Effect of Yiqi Huayu Decoction Combined with Calcium Dobesilate in Treating Diabetic Kidney Disease with Qi Deficiency and Blood Stasis Syndrome and Its Effect on the Expression Levels of Vascular Endothelial Growth Factor and Insulin-like Growth Factor 1 / 广州中医药大学学报

Objective To investigate the clinical efficacy of Yiqi Huayu Decoction(mainly composed of Astragali Radix,Dioscoreae Rhizoma,Poria,fried Euryales Semen,Ecliptae Herba,Rosae Laevigatae Fructus,charred Crataegi Fructus,Ligustri Lucidi Fructus,Salviae Miltiorrhizae Radix et Rhizoma,and Leonuri Herba)combined with Calcium Dobesilate in the treatment of diabetic nephropathy(DN)with qi deficiency and blood stasis syndrome,and to observe the effect of the therapy on vascular endothelial growth factor(VEGF)and insulin-like growth factor 1(IGF-1).Methods Ninety patients with DN of qi deficiency and blood stasis type were randomly divided into an observation group and a control group,with 45 patients in each group.All patients received basic hypoglycemic therapy and treatment for controlling blood pressure and regulating lipid metabolism disorders.Moreover,the patients in the control group were given Calcium Dobesilate orally,and the patients in the observation group were given Yiqi Huayu Decoction combined with Calcium Dobesilate.The course of treatment lasted for 3 months.The changes of traditional Chinese medicine(TCM)syndrome scores,renal function parameters and serum VEGF and IGF-1 levels in the two groups of patients were observed before and after the treatment,and the clinical efficacy of the two groups was evaluated after treatment.Results(1)After 3 months of treatment,the total effective rate of the observation group was 91.11%(41/45),and that of the control group was 75.56%(34/45).The intergroup comparison(tested by chi-square test)showed that the therapeutic effect of the observation group was significantly superior to that of the control group(P＜0.05).(2)After one month and 3 months of treatment,the TCM syndrome scores of both groups were significantly lower than those before treatment(P＜0.05),and the scores after 3 months of treatment in the two groups were significantly lower than those after one month of treatment(P＜0.05).The intergroup comparison showed that the reduction of TCM syndrome scores of the observation group was significantly superior to that of the control group after one month and 3 months of treatment(P＜0.01).(3)After treatment,the levels of renal function parameters such as serum creatinine(Scr),blood urea nitrogen(BUN),and glomerular filtration rate(GFR)in the two groups of patients were significantly improved compared with those before treatment(P＜0.05),and the observation group's effect on the improvement of all renal function parameters was significantly superior to that of the control group(P＜0.01).(4)After treatment,the serum VEGF and IGF-1 levels in the two groups of patients were significantly lower than those before treatment(P＜0.05),and the observation group's effect on the decrease of serum VEGF and IGF-1 levels was significantly superior to that of the control group(P＜0.01).(5)In the course of treatment,no significant adverse reactions occurred in the two groups of patients,with a high degree of safety.Conclusion Yiqi Huayu Decoction combined with Calcium Dobesilate exerts certain therapeutic effect in treating DN patients with qi deficiency and blood stasis syndrome.The combined therapy can effectively down-regulate the serum levels of VEGF and IGF-1,significantly improve the renal function,and alleviate the clinical symptoms of the patients,with a high degree of safety.

miR-146a-3p regulates astrocyte proliferation,migration and apoptosis by inhibiting insulin-like growth factor 1 expression / 中国组织工程研究

BACKGROUND:The alteration of miR-146a-3p level is a common event in the pathogenesis of most neurological diseases,and the specific mechanism of miR-146a-3p regulation of astrocytes has not been studied. OBJECTIVE:To verify that miR-146a-3p regulates astrocyte proliferation,migration and apoptosis through insulin-like growth factor 1. METHODS:12 SD rats were divided into a sham operation group and a spinal cord injury group,with six rats in each group.RNA sequencing analysis was performed on the spinal cord tissues of all groups 2 weeks after surgery to screen out the differential genes(log2FC>2),and to select spinal cord injury-related genes(Score>20)in the Genecards database,and then to predict the target genes of miR-146a-3p by Targetscan.The intersection of three gene sets was obtained to screen out insulin-like growth factor 1 as one of the important target genes.qPCR,western blot assay and immunohistochemistry were performed to analyze the expression level of insulin-like growth factor 1 in spinal cord tissues.The primary astrocytes were divided into NC group,NC-mimics group and miR-146a-3p mimics group.Annexin-V/PI staining was used to detect cell apoptosis.CCK-8 assay was used to detect cell proliferation.Transwell assay was used to detect cell migration ability. RESULTS AND CONCLUSION:The expression of miR-146a-3p in the spinal cord tissue of the spinal cord injury group was lower than that of the sham operation group(P<0.05).The expression of insulin-like growth factor 1 in the spinal cord tissue of the spinal cord injury group was higher than that of the sham operation group(P<0.05).Compared with the NC group and NC-mimics group,the apoptotic rate of astrocytes was increased(P<0.01);the proliferation of astrocytes was decreased(P<0.01)and the number of migration was decreased(P<0.01)in the miR-146a-3p mimics group.To conclude,the expression of miR-146a-3p decreased and the expression of insulin-like growth factor 1 increased in spinal cord tissue after spinal cord injury.miR-146a-3p targeted regulation of insulin-like growth factor 1 in astrocytes,inhibited the proliferation and migration of astrocytes and promoted their apoptosis.

Expression and Clinical Significance of Insulin-like Growth Factor-1 in Pituitary Adenomas / 医学研究杂志

Objective To analyze the expression of insulin-like growth factor-1(IGF-1)in the serum of patients with pituitary adenomas and compare them according to different standards,investigating the expression of IGF-1 in pituitary adenoma and its clinical significance.Methods A total of 156 cases of pituitary adenomas admitted to the Second Affiliated Hospital of Kunming Medical Univer-sity from January 2019 to June 2021 were selected as the experimental group,and a total of 22healthy subjects during the same period were selected as the control group.To determinate the IGF-1 level by magnetic particle chemical luminescence immunoassay.The tumor di-ameter was obtained by combining the diameter measured on magnetic resonance imaging(MRI)with the diameter of postoperative tumor pathological specimen.The experimental group were divided into groups according to different clinical characteristics,and to compare the differences in IGF-1 levels between each subgroup and the control group.Results The levels of IGF-1 in patients with somatotroph adenoma,dual hormone and multihormone adenoma were higher than those in the control group and other types,and the differences were statistically significant(P＜0.05);the level of IGF-1 in giant adenoma patients was higher than those in the control group and other types,and the difference was statistically significant(P＜0.05);the level of IGF-1 in patients with invasive pituitary adenoma was higher than that in non-invasive patients and controls,and the difference was statistically significant(P＜0.05);Logistic regression a-nalysis showed that IGF-1 level and tumor diameter were risk factors for invasiveness of pituitary adenomas,the higher the IGF-1 level and the larger the tumor diameter,the higher the invasiveness risk;the level of IGF-1 in patients with pituitary adenoma within one month after surgery was significantly lower than that before surgery,and the difference was statistically significant(P＜0.05).Conclu-sion IGF-1 has a certain value in the diagnosis of different functional types of pituitary adenomas and the early invasiveness judgment of pituitary adenomas,and is expected to be used as a reliable indicator for postoperative follow-up.

Hedysari Radix Polysaccharide Regulates Apoptosis of Smooth Muscle Cells in Gastric Antrum of Rat Model of Diabetic Gastroparesis via IGF-1/PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effects of Hedysari Radix polysaccharide on the apoptosis of gastric sinus smooth muscle cells and explore the underlying mechanism via the insulin-like growth factor-1 （IGF-1）/phosphatidylinositol 3-kinase （PI3K）/serine-threonine kinase （Akt） pathway in the rat model of diabetic gastroparesis （DGP）. MethodSixty-two Wistar male rats were randomized into a blank group （n=12） and a modelling group （n=50）. The rat model of DGP was established by small-dose multiple intraperitoneal injections of streptozotocin combined with an irregular high-fat and high-sugar diet for 4 weeks. The modeled rats were randomized into model group， mosapride citrate （1.35 mg·kg-1）， and high-， medium-， and low-dose （200， 100， and 50 mg·kg-1， respectively） Hedysari Radix polysaccharide groups. The rats were administrated with corresponding drugs by gavage， and those in the blank and model groups with equal volumes of pure water by gavage once a day for 8 consecutive weeks. The random blood glucose and body mass were measured every 2 weeks， and gastric emptying rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of smooth muscle in gastric antrum， and terminal deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to detect the apoptosis of smooth muscle cells in the gastric antrum. The expression of IGF-1， phosphorylated （p）-PI3K， and p-Akt in the smooth muscle of gastric sinus tissue was detected by immunohistochemistry. Western blot was employed to determine the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the smooth muscle of the gastric antrum. ResultCompared with the blank group， the model group showed elevated random blood glucose at all time points （P<0.01）， decreased body mass and gastric emptying rate （P<0.01）， increased apoptotic index of smooth muscle cells in the gastric antrum （P<0.01）， down-regulated protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated protein level of Bax （P<0.01）. Compared with the model group， the 8 weeks of drug administration lowered the random blood glucose， increased the body mass and gastric emptying rate （P<0.05， P<0.01）， decreased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， up-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and down-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the mosapride citrate group，the administration of low-dose Hedysari Radix polysaccharide for 6 and 8 weeks lowered the random blood glucose and decreased the body mass （P<0.05， P<0.01），low and medium-dose Hedysari Radix polysaccharide decreased the gastric emptying rate and the apoptotic index of smooth muscle cells in the astragaloside low-dose group decreased （P<0.05）. The protein levels of IGF-1，p-PI3K/PI3K，p-Akt/Akt and Bcl-2（low dose）were down-regulated and the protein level of Bax was up-regulated by low doses of Hedysari Radix polysaccharide （P<0.05， P<0.01）. Compared with high-dose Hedysari Radix polysaccharide， low-dose Hedysari Radix polysaccharide elevated random blood glucose and reduced body mass after 6 and 8 weeks of administration （P<0.05， P<0.01）， and the low and medium doses decreased the gastric emptying rate， increased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， down-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the medium-dose group，the low-dose group of Hedysari Radix polysaccharide had lower body mass，lower gastric emptying rate in rats，higher apoptotic index of smooth muscle cells in gastric sinus tissue after 6 and 8 weeks of administration （P<0.05， P<0.01）， and lower protein expression of IGF-1，p-PI3K/PI3K，p-Akt/Akt. ConclusionHedysari Radix polysaccharide protects the smooth muscle cells in gastric antrum against apoptotic injury and promotes gastric motility by activating the IGF-1/PI3K/Akt signaling pathway， as manifested by the up-regulated expression of IGF-1， p-PI3K， p-Akt， and Bcl-2 and down-regulated expression of Bax.

Protective effects of the salt-induced kinase inhibitor HG-9-91-01 on sepsis-associated cognitive dysfunction in mice and the underlying mechanisms / 中南大学学报(医学版)

Objective:Sepsis-associated cognitive dysfunction is a common complication in patients with sepsis and lack of effective treatment.Its pathological mechanisms remain unclear.Salt-induced kinase(SIK)is an important molecule in the regulation of metabolism,immunity,and inflammatory response.It is associated with the development of many neurological diseases.This study aims to investigate the expression of SIK in the hippocampus of septic mice,and to evaluate the role and mechanism of the SIK inhibitor HG-9-91-01 in sepsis-associated cognitive dysfunction. Methods:Firstly,C57BL/6 mice were randomly divided into a control group(Con group)and a sepsis model group[lipopolysaccharide(LPS)group].The model group was injected intraperitoneally with LPS at a dose of 8 mg/kg and the Con group was injected with an equal volume of normal saline.Hippocampal tissues were harvested at 1,3,and 6 days after injection and the expressions of SIK1,SIK2,and SIK3 were detected by real-time fluorescence quantitative PCR(qPCR)and Western blotting.Secondly,C57BL/6 mice were randomly divided into a Con group,a LPS group,and a SIK inhibitor group(HG group).The LPS and HG groups were injected with LPS to establish a sepsis model;in the HG group,HG-9-91-01(10 mg/kg)was injected intraperitoneally at 3-6 days after LPS injection,and the LPS group was injected with the same volume of vehicle.Cognitive function was assessed at 7-11 days after LPS injection using the Morris water maze(MWM).Hippocampal tissues were harvested after the behavioral tests,and the mRNA levels of inflammatory factors and microglial markers were assessed by qPCR.The protein levels of inducible nitric oxide synthase(iNOS),CD68,ionized calcium binding adaptor molecule 1(Iba-1),N-methyl-D-aspartate(NMDA)receptor(NR)subunit,cAMP response element-binding protein(CREB)-regulated transcription coactivator 1(CRTC1),and insulin-like growth factor 1(IGF-1)were detected by Western blotting.Immunohistochemistry(IHC)was used to detect the expression of Iba-1 positive cells in the CA1,CA3 and dentate gyrus(DG)of the hippocampus,followed by Sholl analysis. Results:Compared with the Con group,the mRNA and protein levels of SIK1,SIK2,and SIK3 in the hippocampus were increased in the LPS group(all P<0.05).Compared with the Con group,mice in the LPS group had a significantly longer escape latency,a lower percentage of target quadrant dwell time and a reduced locomotor speed(all P<0.05);the HG group had a decreased escape latency and an increased percentage of time spent in the target quadrant in comparison with the LPS group(both P<0.05).The mRNA levels of inflammatory factors[tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6)],and the M1-type microglial markers iNOS and CD68 in the hippocampus of the LPS group were increased in comparison with the Con group,while the M2-type microglial markers CD206 and arginase-1(Arg-1)were decreased.Compared with the LPS group,the mRNA levels of TNF-α,IL-1β,IL-6,and iNOS were downregulated,while the levels of CD206 and Arg-1 were upregulated in the HG group(all P<0.05).The protein levels of iNOS,CD68,and Iba-1 in the hippocampus of the LPS group were increased in comparison with the Con group,but they were downregulated in the HG group in comparison with the LPS group(all P<0.05).The number of Iba-1 positive cells in CA1,CA3,and DG of the hippocampus was increased in the LPS group in comparison with the Con group,but they were decreased in the HG group in comparison with the LPS group(all P<0.05).Sholl analysis showed that the number of intersections at all radii between 8-38 μm from the microglial soma was decreased in the LPS group in comparison with the Con group(all P<0.05).Compared with the LPS group,the number of intersections at all radii between 14-20 μm was significantly increased in the HG group(all P<0.05).The protein levels of NR subunit NR1,NR2A,NR2B,and IGF-1 were downregulated in the hippocampus of the LPS group in comparison with the Con group,while the expression of phosphorylated CRTC1(p-CRTC1)was increased.Compared with the LPS group,the levels of NR1,NR2A,NR2B,and IGF-1 were upregulated,while p-CRTC1 was downregulated in the HG group(all P<0.05). Conclusion:SIK expression is upregulated in the hippocampus of septic mice.The SIK inhibitor HG-9-91-01 ameliorates sepsis-associated cognitive dysfunction in mice,and the mechanism may involve in the activation of the CRTC1/IGF-1 pathway,inhibition of neuroinflammation,and enhancement of synaptic plasticity.

Progress in the treatment of thyroid-associated ophthalmopathy with teprotumumab / 中华实验眼科杂志

Thyroid-associated ophthalmopathy (TAO) is an autoimmune disease of complex etiology and pathogenesis, which can cause a variety of ocular manifestations and even threaten vision.Glucocorticoids are often used as the first-line treatment in the clinic, but some patients become resistant to them.The insulin-like growth factor-1 receptor (IGF-1R) plays a key role in the development of TAO.Teprotumumab is an anti-IGF-1R monoclonal antibody that can specifically bind to the IGF-1R and block its binding to the α-subunit of IGF-1 to exert biological effects.Clinical trials have shown that teprotumumab has good efficacy in reducing proptosis (decrease≥2 mm) and inflammatory activity (CAS decrease≥2 points) of active moderate-to-severe TAO.Most side effects are mild or moderate.Hyperglycemia is currently an identifiable adverse event associated with teprotumumab that can be controlled with medication.Pregnancy and inflammatory bowel disease are contraindications for teprotumumab due to its teratogenicity to the fetus and its ability to severely exacerbate inflammatory bowel disease.The US FDA has officially approved teprotumumab for the treatment of active moderate-to-severe TAO in January 2020.This article reviews the progress of clinical trials on the mechanism, efficacy and safety of teprotumumab in the treatment of TAO.

Effect of Compound Phyllanthus urinaria Ⅱ on Growth and Expression of miR-122 and IGF-1R of Hepatocellular Carcinoma Hep3B Cells in Nude Mice / 中国实验方剂学杂志

ObjectiveTo investigate the inhibitory effects and mechanism of the compound Phyllanthus urinaria Ⅱ （CPU Ⅱ）on the growth of transplanted hepatocellular carcinoma Hep3B2.1-7 （Short for Hep3R） cells in nude mice. MethodAfter the establishment of a xenograft model of hepatocellular carcinoma Hep3B cells in mice， the model mice were randomly divided into a model group， a high-dose CPU Ⅱ group （57.5 g·kg-1）， a low-dose CPU Ⅱ group （28.75 g·kg-1）， and a 5-fluorouracil （5-FU） group （0.025 g·kg-1）， with eight mice in each group. The mice in the high- and low-dose CPU Ⅱ groups were treated with drugs by gavage， once per day， and those in the model group were treated with the same volume of normal saline. The mice in the 5-FU group were treated by 5-FU by intraperitoneal injection， once every other day. After 28 days of administration， mice were sacrificed， and transplanted tumors were collected. Immunohistochemistry （IHC） was used to detect the expression of proliferating cell nuclear antigen （PCNA） of tumor tissues. Terminal-deoxynucleotidyl transferase-mediated nick end labeling （TUNEL） was used to detect cell apoptosis of tumor tissues. The mRNA expression of miR-122 and insulin-like growth factor 1 receptor （IGF-1R） in tumor tissues was detected by Real-time quantitative PCR （Real-time PCR）. The protein expression of CCAAT/enhancer-binding protein α （C/EBPα）， hepatocyte nuclear factor-4α （HNF-4α）， and IGF-1R in tumor tissues was detected by Western blot. ResultThe tumor suppression rates of the high- and low-dose CPU Ⅱ groups and the 5-FU group were 74.90%， 63.62%， and 64.15%， respectively. Compared with the model group， the CPU Ⅱ groups and the 5-FU group showed reduced weight （P<0.01） and volume of tumors （P<0.01）， decreased PCNA positive cells， shallow staining， increased apoptosis cells of transplanted tumor tissues （P<0.05， P<0.01）， increased expression of mRNA expression of miR-122 （P<0.01）， down-regulated mRNA expression of IGF-1R （P<0.01）， and up-regulated protein expression of C/EBPα and HNF-4α in nude mouse transplanted tumor tissues （P<0.01）. The expression of IGF-1R protein in the high-dose CPU Ⅱ group was down-regulated （P<0.05）. Compared with the low-dose CPU Ⅱ group， the high-dose CPU Ⅱ group showed increased apoptotic cells （P<0.01）， up-regulated mRNA expression of miR-122 （P<0.01）， and increased expression of C/EBPα and HNF-4α proteins （P<0.01）. ConclusionCPU Ⅱ has an obvious inhibitory effect on the growth of transplanted hepatocellular carcinoma Hep3B cells in nude mice. The mechanism of action is related to enhancing the expression of transcription factors HNF-4α and C/EBPα， thereby promoting the expression of miR-122 and inhibiting the expression of its target gene IGF-1R.

Adverse reactions and precautions of teprotumumab in the treatment of thyroid-associated ophthalmopathy / 国际眼科杂志(Guoji Yanke Zazhi)

Thyroid-associated ophthalmopathy(TAO)is an autoimmune disease associated with thyroid dysfunction that can significantly impact quality of life, result in visual impairment and facial disfigurement. Traditional treatments are often unsatisfactory. Studies have shown that teprotumumab, a human monoclonal antibody that can inhibit insulin-like growth factor 1 receptor(IGF-1R), has become an emerging targeted drug for TAO. Although the drug has proven to be effective and relatively safe in the treatment of TAO, adverse reactions are worthy of attention of ophthalmologists with the continuous promotion of clinical application, including hearing impairment, hyperglycemia, diarrhea, muscle spasms, infusion reactions, cognitive decline, thyroid suppression, alopecia, nausea and fatigue. Teprotumumab was generally well tolerated, with most adverse events being mild or moderate in severity. This paper aims to review the adverse reactions and precautions of teprotumumab in the treatment of TAO.

Therapeutic effect of recombinant human growth hormone on children with growth hormone deficiency and different pituitary developmental conditions: a prospective study / 中国当代儿科杂志

OBJECTIVES@#To investigate the therapeutic effect of recombinant human growth hormone (rhGH) on children with growth hormone deficiency (GHD) and different pituitary developmental conditions.@*METHODS@#A prospective study was performed on 90 children with GHD who were admitted to Xuchang Maternity and Child Health Hospital from June 2020 to December 2021. According to pituitary height on the median sagittal plane, they were divided into three groups: pituitary dysplasia group (n=45), normal pituitary group (n=31), and enlarged pituitary growth group (n=14). The changes in body height, growth velocity, height standard deviation score and serum levels of insulin-like growth factor binding protein-3 (IGFBP-3) and insulin-like growth factor-1 (IGF-1) were examined after treatment in the above three groups, and the differences of the above indices before and after treatment were compared among the three groups.@*RESULTS@#After treatment, all three groups had significant increases in body height, growth velocity, height standard deviation score, and the serum levels of IGFBP-3 and IGF-1 (P<0.05). Compared with the normal pituitary group, the pituitary dysplasia group and the enlarged pituitary growth group had significantly higher values in terms of the differences in body height, growth velocity, height standard deviation score, IGF-1, and IGFBP-3 before and after treatment (P<0.05). There was no significant difference in the incidence rate of adverse reactions among the three groups (P>0.05).@*CONCLUSIONS@#In GHD children with different pituitary developmental conditions, rhGH can promote bone growth and increase body height, especially in children with pituitary dysplasia and pituitary hyperplasia, with good safety.

Effect of recombinant human growth hormone on serum Klotho and fibroblast growth factor 23 in children with idiopathic short stature / 中国当代儿科杂志

OBJECTIVES@#To investigate the changes in the serum levels of Klotho, fibroblast growth factor 23 (FGF23), and insulin-like growth factor-1 (IGF-1) in children with idiopathic short stature (ISS) before and after recombinant human growth hormone (rhGH) treatment, as well as the correlation of Klotho and FGF23 with the growth hormone (GH)/IGF-1 growth axis in these children.@*METHODS@#A prospective study was conducted on 33 children who were diagnosed with ISS in the Department of Pediatrics, Hebei Provincial People's Hospital, from March 10, 2021 to December 1, 2022 (ISS group). Twenty-nine healthy children, matched for age and sex, who attended the Department of Child Healthcare during the same period, were enrolled as the healthy control group. The children in the ISS group were treated with rhGH, and the serum levels of Klotho, FGF23, and IGF-1 were measured before treatment and after 3, 6, and 9 months of treatment. A correlation analysis was conducted on these indexes.@*RESULTS@#There were no significant differences in the serum levels of IGF-1, Klotho, and FGF23 between the ISS and healthy control groups (P>0.05). The serum levels of Klotho, FGF23, and IGF-1 increased significantly in the ISS group after 3, 6, and 9 months of rhGH treatment (P<0.05). In the ISS group, Klotho and FGF23 levels were positively correlated with the phosphate level before treatment (P<0.05). Before treatment and after 3, 6, and 9 months of rhGH treatment, the Klotho level was positively correlated with the IGF-1 level (P<0.05), the FGF23 level was positively correlated with the IGF-1 level (P<0.05), and the Klotho level was positively correlated with the FGF23 level (P<0.05), while Klotho and FGF23 levels were not correlated with the height standard deviation of point (P>0.05).@*CONCLUSIONS@#The rhGH treatment can upregulate the levels of Klotho, FGF23, and IGF-1 and realize the catch-up growth in children with ISS. Klotho and FGF23 may not directly promote the linear growth of children with ISS, but may have indirect effects through the pathways such as IGF-1 and phosphate metabolism. The consistent changes in Klotho, FGF23 and IGF-1 levels show that there is a synergistic relationship among them in regulating the linear growth of ISS children.

Diagnostic significance and considerations of growth hormone stimulation testing and insulin-like growth factor 1 in growth hormone deficiency / 中国当代儿科杂志

The growth hormone (GH)/insulin-like growth factor 1 (IGF-1) axis is an essential component of the hypothalamic-pituitary growth hormone axis and plays a crucial role in childhood growth and development. Disruptions and abnormalities in the GH/IGF-1 signaling pathway and its pathways typically manifest as short stature in children. Children with short stature often undergo GH stimulation testing and IGF-1 level measurements to differentiate growth hormone deficiency (GHD) from other causes of growth delay. This article aims to analyze and elucidate the values of GH stimulation testing and IGF-1 measurement, providing reference for the diagnosis of GHD in children.

Effects of p,p'-DDE on H19 DMR methylation and insulin secretion of INS-1 cells / 环境与职业医学

Background Diabetes is a major threat to public health across the world. Studies have shown that exposure to p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) is closely related to the occurrence of type 2 diabetes mellitus. However, the relevant molecular mechanism is not clear. Objective To investigate the effects of p,p'-DDE on H19 differentially methylated region (DMR) methylation and insulin secretion of rat insulinoma cells (INS-1 cells). Methods INS-1 cells were cultured with different concentrations (0, 3.125, 6.25, 12.5, 25, 50, and 75 µmol·L−1) of p,p'-DDE for 24 h, and the viability of INS-1 cells was detected by CCK-8 method. INS-1 cells were exposed to 0, 12.5, 25, and 50 µmol·L−1 p,p'-DDE for 24 h in subsequent experiments. The methylation levels of 24 CpG sites in H19 DMR were analyzed by bisulfite genomic sequencing. The expression levels of insulin-like growth factor 2 (IGF2) mRNA were detected by real-time quantitative PCR. The expression levels of IGF2 and insulin-like growth factor-1 receptor (IGF1R) proteins were detected by Western blotting. The insulin secretion function of INS-1 cells was determined by glucose-stimulatedinsulin secretion test (5 and 25 mmol·L−1 glucose, respectively). Results Compared with the control group, the viability of INS-1 cells increased significantly after treatment with 12.5 µmol·L−1 p,p'-DDE; however, it was significantly inhibited after treatment with 50 or 75 µmol·L−1 p,p'-DDE (P<0.01); therefore, 50 µmol·L−1 was chosen as the maximum concentration of exposure for subsequent experiments. The 25 µmol·L−1 p,p'-DDE treatment decreased the methylation levels of CpG18 and CpG22-CpG24 sites in H19 DMR, and the 50 µmol·L−1 p,p'-DDE treatment decreased the methylation levels of CpG10-CpG24 sites (P<0.05 or P<0.05). Multiple concentrations (12.5, 25, and 50 µmol·L−1) of p,p'-DDE down-regulated the mRNA and protein relative expression levels of IGF2 and the protein relative expression levels of IGF1R. The transcription level of IGF2 decreased to 67.8%, 68.6%, and 62.5% of the control group, the protein level of IGF2 decreased to 73.3%, 79.5%, and 80.9% of the control group, and the protein level of IGF1R decreased to 54.8%, 25.6%, and 12.9% of the control group, respectively (P<0.01). In the high glucose context, p,p'-DDE at selected concentrations inhibited the insulin secretion levels to 85.0%, 58.6%, and 49.5% of the control group, respectively (P<0.01). Conclusion p,p'-DDE could down-regulate methylation level of H19 DMR, interfere the IGF2/IGF1R signaling pathway, and inhibit insulin secretion of islet cells.

Effect of milk on sebaceous gland spots and the IGF-1/SREBP-1/ACC-1 signaling pathway in golden hamsters / 中华皮肤科杂志

Objective:To investigate the effect of milk on sebum secretion in golden hamsters, and to explore its possible mechanism of action.Methods:Eighteen golden hamsters were randomly and equally divided into 3 groups: blank control group receiving no intervention, whole-milk group gavaged with whole milk, and skimmed-milk group gavaged with skimmed milk. The gavage feeding was performed at a dose of 2.5 ml twice a day for 4 consecutive weeks. The maximum transverse diameter and maximum longitudinal diameter of bilateral sebaceous gland spots were measured on days 0, 7, 14, 21 and 28 after the start of intervention, and the area of sebaceous gland spots was calculated; at 24 hours after the last gavage, bilateral sebaceous gland spot tissues were resected, and subjected to immunohistochemical study to determine the expression of insulin-like growth factor-1 (IGF-1) /sterol regulatory element-binding protein-1 (SREBP-1) /acetyl-coenzyme A carboxylase (ACC-1) signaling pathway in sebaceous gland spots. Statistical analysis was carried out by using repeated measures analysis of variance, one-way analysis of variance for independent groups, Kruskal-Wallis H test, and least significant difference- t test for multiple comparisons. Results:Repeated measures analysis of variance showed that there was no significant difference in the area of sebaceous gland spots of golden hamsters among the 3 groups ( F= 0.96, P= 0.417) . The IGF-1 expression was significantly higher in the skimmed-milk group (0.39 ± 0.03) than in the blank control group (0.35 ± 0.03, t= 2.62, P= 0.021) and whole-milk group (0.33 ± 0.02, t= 3.82, P= 0.002) ; compared with the blank control group (0.36 ± 0.02) , the skimmed-milk group showed significantly increased SREBP-1 expression (0.42 ± 0.04, t= 2.64, P= 0.021) ; the ACC-1 expression was significantly higher in the skimmed-milk group (0.40 ± 0.03) and whole-milk group (0.40 ± 0.05) than in the blank control group (0.34 ± 0.03; t= 2.39, 2.47, P= 0.031, 0.026, respectively) . Conclusion:Milk may promote sebum secretion in golden hamsters through the IGF-1/SREBP-1/ACC-1 signaling pathway.

Research progress on the relationship between Klotho and the growth of children and newborns / 中国综合临床

Klotho is a gene associated with aging, the transmembrane protein encoded by this gene is highly expressed in the kidney, and is also expressed in tissues such as the brain, parathyroid and pituitary glands. The extracellular domain of Klotho can also be cleaved and shed to form soluble Klotho, which acts as a circulating hormone and can be detected in blood, cerebrospinal fluid, and urine. More and more studies have shown that Klotho protein plays an important role in the complex regulation of growth hormone (GH)-insulin-like growth factor 1(IGF1) axis. The interaction between Klotho protein and GH-IGF1 axis is bidirectional, which regulates each other, and then regulates the normal linear growth of children. In addition, Klotho protein can also affect the growth and development of fetus and newborn through different ways, and its mechanism is not very clear.

Effect of IGF1Rβ Subunit Mutants on Proliferation, Migration and Apoptosis of Human Osteosarcoma Cells / 肿瘤防治研究

Objective To investigate the effect of IGF1R β subunit mutants sb-IGF1R and ma-IGF1R on the biological behavior of osteosarcoma 143B cells. Methods We designed and constructed sb-IGF1R and ma-IGF1R fragments. They were cloned into adenovirus AdEasy shuttle plasmid, to obtain Ad-sbIGF1R and Ad-maIGF1R. We observed the proliferation, migration and apoptosis of the osteosarcoma cells transfected with Ad-sbIGF1R, Ad-maIGF1R and Ad-IGF1R. The Ad-sbIGF1R, Ad-maIGF1R and Ad-GFP nude mouse models were constructed to evaluate the tumor growth in vitro. Results By plasmid PCR, IGF-1R β subunit mutant was overexpressed in osteosarcoma cells. Ad-sbIGF1R and Ad-maIGF1R significantly inhibited the proliferation and migration of osteosarcoma cells, and promoted cell apoptosis, and inhibited tumor growth in subcutaneous tumor-bearing nude mouse models. Conclusion IGF1R β subunit mutants inhibit the proliferation and migration of osteosarcoma cells and induce cell apoptosis.

Serum levels of insulin-like growth factor-1 and insulin-like growth factor binding protein-3 in children with autism spectrum disorder / 中国当代儿科杂志

OBJECTIVES@#To study the serum levels of insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-3 (IGFBP-3) in children with autism spectrum disorder (ASD) and their association with the core symptoms of ASD.@*METHODS@#A total of 150 ASD children aged 2-7 years (ASD group) and 165 healthy children matched for age and sex (control group) who were recruited at the outpatient service of Chongqing Health Center for Women and Children were enrolled as subjects. Autism Behavior Checklist (ABC) and Childhood Autism Rating Scale (CARS) were used to evaluate the core symptoms of the ASD children. Chemiluminescence was used to measure the serum levels of IGF-1 and IGFBP-3 in both groups.@*RESULTS@#The ASD group had a significantly lower serum level of IGF-1 than the control group (P<0.05). The children with severe ASD had significantly lower serum levels of IGF-1 and IGFBP-3 than those with mild-to-moderate ASD (P<0.001). For the children aged 2-3 years, the ASD group had a significantly lower serum level of IGF-1 than the control group (P<0.05). Boys had a significantly lower serum level of IGF-1 than girls in both ASD and control groups (P<0.05). The serum levels of IGF-1 and IGFBP-3 were negatively correlated with the total score of CARS (r=-0.32 and -0.40 respectively, P<0.001).@*CONCLUSIONS@#The reduction in serum IGF-1 level in early childhood may be associated with the development of ASD, and the serum levels of IGF-1 and IGFBP-3 are associated with the core symptoms of ASD children.

Repercussão da adenotonsilectomia sobre o estado nutricional em crianças respiradoras orais / Repercussion of adenotonsillectomy on the nutritional status of mouth breathing children

Introdução: a hiperplasia adenotonsilar (HAT) é uma das causas mais comuns da Síndrome do Respirador Oral (SRO) devido à obstrução de via aérea superior em crianças e adolescentes. Tal afecção pode causar alterações ortodônticas, miofuncionais orofaciais, posturais, cardiopulmonares, antropométricas e polissonográficas. O diagnóstico precoce e indicação de Adenotonsilectomia (A&T) é essencial para reversão dessas consequências deletérias da SRO e restauração do bem estar biopsicossocial da criança.Objetivo: avaliar o estado nutricional, patência nasal, distúrbios do sono e fator de crescimento semelhante à insulina tipo 1 (IGF-1) em crianças de dois a doze anos de idade com SRO devido HAT grave e comparar com a reavaliação após seis meses de pós-cirúrgico das crianças operadas e com as demais que permanecem com obstrução da via aérea e aguardam a cirurgia na fila de espera do Sistema Único de Saúde. Métodos: trinta pacientes com SRO por HAT grave e indicação de A&T foram submetidos à avaliação antropométrica, polissonográfica, dosagem do IGF-1, rinomanométrica, teste alérgico cutâneo, questionário de padrão alimentar e prática de atividade física antes da A&T. Dez pacientes repetiram essa avaliação seis meses após o procedimento cirúrgico (grupo intervenção). Vinte pacientes aguardam a cirurgia na fila de espera do SUS e tiveram seus dados antropométricos e de IGF-1 reavaliados após seis meses com obstrução da via aérea (grupo controle). Resultados: trinta crianças realizaram a fase pré-operatória do estudo. A idade média foi de 5,6 anos (±2,17). Dezessete (56,7%) eram do sexo masculino e treze (43,3%) do sexo feminino. O teste cutâneo foi positivo em dezesseis indivíduos (53,3%) As médias dos escores Z de estatura por idade foi de -0,95 (±1,09); peso por idade de 0,17 (±1,42); índice de massa corporal (IMC) por idade de 0,31 (±1,36). A média do fluxo nasal inspiratório total (FNIT) foi de 444,63 ml/s (±161,02) e da patência nasal de 72,9% (±24,76). A média do índice de Apneia e Hipopneia (IAH) do sono foi de 4,95 ev/h (±4,07); da saturação mínima de oxihemoglobina no sono (Nadir de O2) de 78,93% (±6,00); da percentagem de sono com saturação menor que 90% (T90) de 4,16% (±5,48); da porcentagem do sono com ondas lentas (sono N3) de 37,62% (±9,61). A média do escore Z de IGF-1 foi de 0,72 (±1,30). O grupo intervenção e grupo controle não apresentaram alterações dos dados antropométricos com significância estatística. Houve diminuição do IGF-1 após a cirurgia sendo a média do escore Z de IGF-1 pré-operatório de 1,33 (±1,74) e pós-cirúrgico de -0,07 (±0,85); p=0,03. No grupo controle a variação do IGF-1 não foi significativa. O grupo intervenção não apresentou alteração com significância estatística do FNIT e da patência nasal. Nas dez crianças operadas foi constatada uma melhora da média do IAH de 5,25 ev/h (±4,29) para 1,99 ev/h (±1,16) e do T90 de 6,27% (±7,46) para 0,64% (±0,55) com p<0,05. Já o sono N3 e o Nadir de O2 não apresentaram alterações significativas. Não houve mudança qualitativa no padrão alimentar e na prática de atividade física nos dois períodos avaliados na vigência da pandemia de COVID19. Conclusão: Após A&T houve diminuição do IGF-1; p=0,03, melhora do IAH; p=0,03 e do T90; p=0,04. A cirurgia não modificou o estado nutricional com significância estatística nas dez crianças após 6 meses de pós-operatório. No pós-cirúrgico, não houve diferença estatística do FNIT e da patência nasal, assim como nessa amostra também não ocorreram alterações significativas do sono N3 e do Nadir de O2. O padrão alimentar e a prática de atividade física foram semelhantes qualitativamente no pré e no pós-operatório. Vinte crianças no grupo controle não tiveram alterações significativas dos dados antropométricos e do IGF-1 com seis meses de espera pela cirurgia e permanência da obstrução da via aérea. Não houve diferença estatística dos dados antropométricos e do IGF-1 entre o grupo controle e o grupo intervenção.

Introduction: adenotonsillar hyperplasia (ATH) is one of the most common causes of Mouth Breathing Syndrome (MBS) due to upper airway obstruction in children and adolescents. This condition can cause orthodontic, orofacial myofunctional, postural, cardiopulmonary, anthropometric and polysomnographic changes. Early diagnosis and indication of Adenotonsillectomy (T&A) is essential to revert these deleterious consequences of MBS and restore the child's biopsychosocial well-being. Objective: to evaluate the nutritional status, nasal patency, sleep disorders and insulin-like growth factor 1 (IGF-1) in children aged two to twelve years old with MBS due to severe ATH and compare with reassessment after six months post-surgical care of operated children and others who remain with airway obstruction and are waiting for surgery on the Unified Health System (UHS) waiting list. Methods: Thirty patients with MBS due to severe ATH and indication for T&A were submitted to anthropometric, polysomnographic, IGF-1 dosage, rhinomanometric, allergic skin test, dietary pattern questionnaire and physical activity practice before T&A. Ten patients repeated this evaluation six months after the surgical procedure (intervention group). Twenty patients were waiting for surgery on the UHS waiting list and had their anthropometric and IGF-1 data reassessed after six months with airway obstruction (control group). Results: Thirty children underwent the preoperative phase of the study. The mean age was 5.6 years (±2.17). Seventeen (56.7%) were male and thirteen (43.3%) were female. The skin test was positive in sixteen individuals (53.3%) The average Z-scores for height for age were -0.95 (±1.09); weight for age 0.17 (±1.42); body mass index (BMI) for age of 0.31 (±1.36). The mean total inspiratory nasal flow (TINF) was 444.63 ml/s (±161.02) and nasal patency was 72.9% (±24.76). The average sleep apnea and hypopnea index (AHI) was 4.95 ev/h (±4.07); minimum oxyhemoglobin saturation during sleep (O2 Nadir) of 78.93% (±6.00); percentage of sleep with saturation lower than 90% (T90) of 4.16% (±5.48); percentage of sleep with slow waves (N3) of 37.62% (±9.61). The mean IGF-1 Z-score was 0.72 (±1.30). The intervention group and control group did not show statistically significant changes in anthropometric data. There was a decrease in IGF-1 after surgery, with a mean preoperative IGF-1 Z-score of 1.33 (±1.74) and postoperative value of -0.07 (±0.85); p=0.03. In the control group, the IGF-1 variation was not significant. The intervention group did not show statistically significant changes in TINF and nasal patency. In the ten operated children, an improvement in the mean AHI from 5.25 ev/h (±4.29) to 1.99 ev/h (±1.16) and T90 of 6.27% (±7. 46) to 0.64% (±0.55) with p<0.05. On the other hand, N3 sleep and O2 Nadir showed no significant changes. There was no qualitative change in dietary patterns and physical activity in the two periods evaluated during the COVID19 pandemic. Conclusion: After T&A there was a decrease in IGF-1; p=0.03, AHI improvement; p=0.03 and T90 too; p=0.04. The surgery did not change the nutritional status with statistical significance in the ten children after 6 months postoperatively. Post-surgery, there was no statistical difference in TINF and nasal patency, as well as in this sample there were no significant changes in N3 sleep and O2 Nadir either. The dietary pattern and the practice of physical activity were qualitatively similar before and after the operation. Twenty children in the control group did not have significant alterations in anthropometric data and IGF-1 after six months of waiting for the surgery and the remaining airway obstruction. There was no statistical difference in anthropometric and IGF-1 data between the control and intervention groups.

Rehabilitation therapy with insulin-like growth factor after acute skeletal muscle injury / Terapia de reabilitação com fator de crescimento semelhante à insulina após lesão muscular esquelética aguda / Terapia de rehabilitación con factor de crecimiento semejante a la insulina después de una lesión aguda del músculo esquelético

ABSTRACT Introduction Skeletal muscle injuries account for 10% to 50% of treadmill sports injuries. Insulin-like growth factor (IGF) is a family of polypeptides with both insulin-like anabolic and growth-promoting effects. Sports play a vital role in the recovery of skeletal muscle injuries. Objective The paper analyzes the ability of insulin-like growth factor 1 (IGF-1) to repair skeletal muscle injury caused by treadmill exercise. Method We injected drugs under the wound after exercise-induced injury in rats. The control group was injected with saline, and the experimental group was injected with an insulin-like growth factor. We conduct histological and electron microscopic structural analysis of rats, Results: After an injury, the experimental group formed a basal lamina protective film earlier than the control group, activated myoblasts, formed myofilaments, formed myotubes, and fused into muscle fibers earlier than the control group. The healing quality was also better. The experimental group was endogenous. The mRNA content of sex IGF-1 and IGF-2 both increased earlier than the control group. Conclusion Local injection of exogenous insulin-like growth factor-1 can stimulate the proliferation of myoblasts and accelerate the post-traumatic repair process of skeletal muscle caused by treadmill sports. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução As lesões do músculo esquelético representam de 10% a 50% das lesões em esteira esportiva. O fator de crescimento semelhante à insulina (IGF) é uma família de polipeptídeos com efeitos anabólicos e de promoção do crescimento semelhantes à insulina. Os esportes desempenham um papel vital na recuperação de lesões musculares esqueléticas. Objetivo o artigo analisa a capacidade do fator de crescimento semelhante à insulina 1 (IGF-1) em reparar lesões musculares esqueléticas causadas por exercícios em esteira. Método Injetamos drogas sob a ferida após lesão induzida por exercício em ratos. O grupo controle foi injetado com solução salina e o grupo experimental foi injetado com um fator de crescimento semelhante à insulina. Realizamos análises histológicas e microscópicas eletrônicas estruturais de ratos. Resultados Após a lesão, o grupo experimental formou um filme protetor da lâmina basal mais cedo do que o grupo controle, mioblastos ativados, miofilamentos formados, miotubos formados e fundidos em fibras musculares mais cedo do que o grupo controle. A qualidade da cura também foi melhor. O grupo experimental era endógeno. O conteúdo do sexo IGF-1 e IGF-2 mRNA aumentou mais cedo do que no grupo de controle. Conclusão A injeção local de fator de crescimento semelhante à insulina 1 exógeno pode estimular a proliferação de mioblastos e acelerar o processo de reparo muscular esquelético pós-traumático causado por esportes em esteira. Nível de evidência II; Estudos terapêuticos: investigação dos resultados do tratamento.

RESUMEN Introducción Las lesiones del músculo esquelético representan del 10% al 50% de las lesiones deportivas en cinta. El factor de crecimiento semejante a la insulina (IGF) es una familia de polipéptidos con efectos anabólicos y estimulantes del crecimiento semejantes a la insulina. Los deportes juegan un papel vital en la recuperación de las lesiones del músculo esquelético. Objetivo El artículo analiza la capacidad del factor de crecimiento semejante a la insulina 1 (IGF-1) para reparar la lesión del músculo esquelético causada por el ejercicio en cinta. Método inyectamos drogas debajo de la herida después de una lesión inducida por el ejercicio en ratas. Al grupo de control se le inyectó solución salina y al grupo experimental se le inyectó un factor de crecimiento semejante a la insulina. Realizamos análisis estructurales histológicos y microscópicos electrónicos de ratas, Resultados: Después de una lesión, el grupo experimental formó una película protectora de la lámina basal antes que el grupo de control, activó mioblastos, formó miofilamentos, formó miotubos y se fusionó en fibras musculares antes que el grupo de control. La calidad de curación también fue mejor. El grupo experimental fue endógeno. El contenido de ARNm de IGF-1 e IGF-2 de sexo aumentaron antes que en el grupo de control. Conclusión La inyección local de factor de crecimiento semejante a la insulina 1 exógeno puede estimular la proliferación de mioblastos y acelerar el proceso de reparación postraumático del músculo esquelético causado por los deportes en cinta. Nivel de evidencia II; Estudios terapéuticos: investigación de los resultados del tratamiento.

Effects of vitamin e on the epithelial-mesenchymal transition in testicular development exposed to valproic acid / Efectos de la vitamina e en la interacción epitelio-mesenquimático en el desarrollo testicular expuesto a ácido valproico

SUMMARY: In testicular differentiation, somatic cells must adopt a specific destiny towards sustentacular, peritubular and interstitial cells, being fundamental for the morphogenesis of seminiferous tubules, mediated by morphogens such as Desert Hedgehog (DHH), insulin-like growth factor-1 (IGF-1) and fibroblastic growth factor 2 (FGF-2). Its alteration could be related to failures in the development mechanisms, such as those caused by valproic acid (VPA), which can be reversed with vitamin E (VE). The objective of the study was to evaluate the epithelial-mesenchymal transition (EMT) in the testicular development of mice exposed to VPA and VE. 12 groups of pregnant female mice were formed that were separated by days post-coital (dpc) at 12.5 dpc, 17.5 dpc and 6 weeks postnatal, each one subdivided into 4 groups of 5 pregnant women each. Subgroups received different treatments from the beginning to the end of gestation orally: 600 mg/kg of VPA, 600 mg/kg of VPA and 200 IU of VE, 200 IU of VE and the control group 0.3 mL of 0.9% physiological solution. Immunohistochemistry was performed for the detection of DHH, IGF-1 and FGF-2. Immunolocalization of DHH was observed in all stages, with more evident significant differences in integrated optical density (IOD) and percentage of immunoreaction area at 6 weeks postnatal, being lower in the VPA group. In IGF-1, lower intensity and distribution of immunostaining was observed in the fetal and pubertal stages in the VPA groups, a similar situation with FGF-2, but only evident at 17.5 dpc, with significant differences. These results demonstrate that VPA can alter EMT between somatic cells in testicular development, with VE being an agent capable of attenuating this process.

RESUMEN: En la diferenciación testicular, es necesario que las células somáticas adopten un destino específico hacia células sustentaculares, peritubulares e intersticiales, siendo fundamental para la morfogénesis de los túbulos seminíferos, mediado por morfógenos como Desert Hedgehog (DHH), Factor de Crecimiento Fibroblástico 2 (FGF-2) y Factor de Crecimiento símil a Insulina (IGF-1). Su alteración se podría relacionar a fallas en los mecanismos de desarrollo, como los que ocasiona el ácido valproico (VPA), los cuales pueden ser revertidos con la vitamina E (VE). El objetivo de estudio fue evaluar la transición epitelio-mesenquimática (EMT) en el desarrollo testicular de ratones expuestos a VPA y VE. Se conformaron 12 grupos de ratones hembra gestantes que se separaron por días post-coital (dpc) a los 12.5 dpc, 17.5 dpc y 6 semanas post-natal, cada uno subdividido en 4 grupos de 5 gestantes cada uno. Cada subgrupo recibió diferentes tratamientos desde el inicio hasta el término de la gestación vía oral: 600 mg/kg de VPA, 600 mg/kg de VPA y 200 UI de VE, 200 UI de VE y el grupo control 0,3 mL de solución fisiológica 0,9%. Se realizó técnica inmunohistoquímica para la detección de DHH, IGF-1 y FGF-2. Se observó la inmunolocalización de DHH en todos los estadios, con diferencias significativas más evidentes en la densidad óptica integrada (IOD) y porcentaje de área de inmunoreacción a las 6 semanas post-natal, siendo menor en el grupo VPA. En IGF-1, se observó en la etapa fetal y puberal menor intensidad y distribución de la marcación en los grupos VPA, situación similar con la inmunomarcación de FGF-2, pero sólo evidenciándose a los 17.5 dpc, con diferencias significativas. Estos resultados demuestran que el VPA puede alterar la EMT entre las células somáticas en el desarrollo testicular, siendo la VE un agente capaz de atenuar este proceso.

Antidepressant Effect of n-Butyl Alcohol Extracts from Xiaoyaosan via IGF-1Rβ/PI3K/Akt Pathway / 中国实验方剂学杂志

Objective:To investigate the intervention effect of <italic>n</italic>-butyl alcohol extracts from Xiaoyaosan against depression-like behavior induced by chronic unpredictable mild stress （CUMS） in model mice and the role of insulin-like growth factor-1 receptor <italic>β</italic> （IGF-1R<italic>β</italic>）/phosphatidylinositol-3 kinase （PI3K）/protein kinase B （Akt） signaling pathway in such intervention. Method:The effective dose of n-butyl alcohol extracts from Xiaoyaosan was preliminarily determined in model mice with behavioral despair. Then the male C57BL/6 mice were randomly divided into the blank group， model group， fluoxetine group， Xiaoyaosan group， and the low- （20 g·kg^-1） and high-dose （40 g·kg^-1） <italic>n</italic>-butyl alcohol extract groups. The mice in all groups except for the blank group were exposed to CUMS for inducing the depression-like behavior， which was judged by the sucrose preference test （SPT）. The successfully modeled mice in the medication groups were intragastrically administered with the corresponding drugs， whereas those in the blank and model groups were treated with an equal volume of solvent for five successive weeks. Following the SPT， tail suspension test （TST）， and novelty suppressed feeding test （NSFT） at the end of the fifth week， the insulin-like growth factor-1 （IGF-1） levels in mouse serum and hippocampus were measured by enzyme-linked immunosorbent assay （ELISA）. The average optical density （<italic>IA</italic>） of Nissl bodies in mouse hippocampal CA3 region was detected by toluidine blue staining. The 5-bromo-2-deoxyuridine （Brdu） and doublecortin （DCX） expression in the dentate gyrus （DG） was assayed using immunofluorescence method. The protein expression levels of IGF-1R<italic>β</italic>， PI3K， phosphorylated-PI3K （p-PI3K）， Akt， p-Akt， cysteine aspartic acid-specific protease 3 （Caspase-3）， and cleaved Caspase-3 in the hippocampus were determined by Western blot. Result:The results of forced swimming test and TST showed that n-butyl alcohol extracts from Xiaoyaosan at 9.1 and 40 g·kg^-1 both significantly shortened the immobility time of mice （<italic>P</italic><0.05， <italic>P</italic><0.01）， indicating that the effective dose ranged from 9.1-40 g·kg^-1. Compared with the model control， the n-butyl alcohol extracts from Xiaoyaosan at 20 and 40 g·kg^-1 significantly increased the sucrose preference percentage （<italic>P</italic><0.05， <italic>P</italic><0.01）， shortened the immobility time in TST （<italic>P</italic><0.01） and the feeding latency in NSFT （<italic>P</italic><0.01）， reversed the down-regulated IGF-1 content in mouse serum and hippocampus （<italic>P</italic><0.01）， increased the AOD of Nissl bodies in the hippocampal CA3 region （<italic>P</italic><0.01）， promoted the expression of Brdu and DCX in DG （<italic>P</italic><0.05， <italic>P</italic><0.01）， and down-regulated the protein expression levels of IGF-1R<italic>β</italic> （<italic>P</italic><0.05， <italic>P</italic><0.01）， p-PI3K/PI3K （<italic>P</italic><0.05， <italic>P</italic><0.01）， p-Akt/Akt （<italic>P</italic><0.05）， and cleaved Caspase-3/Caspase-3 in the hippocampus of CUMS mice. Conclusion:The n-butyl alcohol extracts from Xiaoyaosan are equivalent to Xiaoyaosan in inhibiting expression. They alleviate the depression-like behavior in CUMS mice， induce the production of Nissl bodies in hippocampal CA3 region， enhance neuronal proliferation and differentiation in DG， and facilitate neurogenesis. All these may be related to the inhibition of over-activated IGF-1R<italic>β</italic>/PI3K/Akt pathway and the reduction of neuronal apoptosis.