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1.
Food Chem ; 368: 130816, 2022 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-34416489

RESUMEN

Acrylamide (AA), a potential carcinogen, is commonly formed in foods rich in carbohydrates at high heat. It is known that AA-induced mitochondrial dysfunction is responsible for its toxicity. Previously we found AA exposure increased miR-27a-5p expression in livers of SD rats. Here, the regulation mechanism of miR-27a-5p in mitochondrial dysfunction was investigated in rat liver cell lines (IAR20) and SD rats. The results showed that the overexpressed miR-27a-5p contributes to modulating mitochondrial dysfunction and Btf3 is identified as its target gene. The knockdown of Btf3 increases the cleaved PARP1 level and the phosphorylation of ATM and p53, which results in mitochondria-dependent apoptosis. Therefore, the miR-27a-5p-Btf3-ATM-p53 axis might play a vital role in the promotion of AA-induced cell apoptosis through disrupting mitochondrial structure and function. This would provide a potential target for the assessment and intervention of AA toxicity.


Asunto(s)
MicroARNs , Acrilamida/toxicidad , Animales , Apoptosis , MicroARNs/genética , Mitocondrias/genética , Ratas , Ratas Sprague-Dawley
2.
J Pharm Biomed Anal ; 207: 114431, 2022 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-34710728

RESUMEN

Sinapine thiocyanate (ST) is an index component and pharmacological active component of Semen Sinapis and Semen Raphani, and it is widely used to relieving cough and asthma. This study aimed to obtain the metabolic and pharmacokinetic characterization of ST. The metabolic profiles of ST were obtained from rat plasma, urine, and feces via ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-Q/TOF-MS). Thirteen metabolites were structurally identified, and the proposed metabolic pathways of ST included deamination, demethylation, hydrogenation, dehydration, and extensive conjugation, including glucuronidation and sulfonation. ST was selected as the plasma marker for the pharmacokinetic study. A simple and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantitation of ST in rat plasma. The linear range of ST was 0.1-500 ng/mL (R2 = 0.9976), and the lowest limit of quantification was 0.1 ng/mL. The intra-precision and inter-precision of the assay were 1.31-5.12% and 2.72-7.66%, and the accuracy (RE%) ranged from - 4.88% to 3.82% and - 3.47% to 6.18%. The extraction recovery, matrix effect, and stability of ST were within acceptable limits. The established method was validated and successfully applied to the pharmacokinetic study of ST. For pharmacokinetic experiments, the male Sprague-Dawley rats were administrated with ST solution intravenously (2 mg/kg) or orally (100 mg/kg). The oral absolute bioavailability of ST was calculated as 1.84%, and the apparent volume of distribution of intravenous and intragastric administrations were 107.51 ± 21.16 L/kg and 78.60 ± 14.44 L/kg, respectively. The maximum plasma concentration was 47.82 ± 18.77 nM, and the time to maximum peak was 88.74 ± 20.08 min for the intragastric dosing group. According to the pharmacokinetic and metabolic profiling results, metabolites with high abundance of ST in bio-fluids would be the next object in tissue distribution and pharmacodynamic study.


Asunto(s)
Espectrometría de Masas en Tándem , Tiocianatos , Administración Oral , Animales , Colina/análogos & derivados , Cromatografía Líquida de Alta Presión , Masculino , Ratas , Ratas Sprague-Dawley
3.
Biol Trace Elem Res ; 200(1): 228-237, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33566284

RESUMEN

Organophosphate compounds are the most widely employed insecticides in countries with high agriculture activity. On average, organophosphates cause 3 million people to poison and 200 000 deaths per year due to food chain or occupational, accidental, or suicidal exposure. Our study aimed to research selenium's protective role against the toxic action of CPF, one of the most commonly used organophosphates, with an experimental model formed with rats. A total of 56 male SD rats were distributed into seven groups as follows: control (tap water), sham (corn oil), group I (5.4 mg/kg CPF), group II (13.5 mg/kg CPF), group III (3 mg/kg Se), group IV (5.4 mg/kg CPF+Se), and group V (13.5 mg/kg CPF+Se). Following 6 weeks of oral exposure, there were significant changes in AChE activity, biochemical and hematological parameters, and trace element levels in CPF-treated rats. In the high-dose CPF group, RBC values, Hb, and Hct decreased, and values of WBC, AST, ALT, ALP increased (p < 0.001) significantly compared to control, sham, and Se groups. While there was no significant change in zinc level, the copper and selenium levels were significantly higher in group IV than in control (p < 0.001) and sham (p < 0.05, p < 0.01, respectively) groups. Moreover, max. O.R.L. was found statistically more elevated in the high-dose CPF group compared to control, sham, and Se groups (p < 0.05, p < 0.05, and p < 0.01, respectively). All results indicated that Se is an antioxidant that reduces the toxic effects caused by CPF. Employing combinations of chlorpyrifos and selenium appeared greatly in restoring the harmful effects of CPF exposure.


Asunto(s)
Cloropirifos , Insecticidas , Selenio , Oligoelementos , Animales , Cloropirifos/toxicidad , Insecticidas/toxicidad , Masculino , Ratas , Ratas Sprague-Dawley , Selenio/farmacología
4.
Biol Trace Elem Res ; 200(1): 271-280, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33629228

RESUMEN

Excessive fluoride (F) exposure can lead to liver damage; moreover, recent studies found that the addition of appropriate calcium (Ca) can alleviate the symptom of skeletal fluorosis. However, whether Ca can relieve F-induced liver damage through the mitochondrial apoptosis pathway has not been reported yet. Therefore, we assessed the liver morphology, serum transaminase content, liver oxidative stress-related enzymes, and apoptosis-related gene and protein expression in Sprague Dawley (SD) rats treated with 150 mg/L sodium fluoride (NaF) and different concentrations of calcium carbonate (CaCO3) for 120 days. Our results showed that NaF brought out pathological changes in liver morphology, serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels increased, total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) content decreased, and malondialdehyde (MDA) content increased, suggesting that NaF caused hepatotoxicity and oxidative stress. In addition, the results of quantitative real-time PCR (qRT-PCR) and immunohistochemistry showed that NaF exposure upregulated the expression of Bcl-2-associated x protein (Bax), rho-related coiled-coil kinase 1 (ROCK1), cytochrome C (Cyto-C) mRNA and protein (P < 0.01), and downregulated B cell lymphoma 2 (Bcl-2) protein and mRNA (P < 0.01), indicating that excessive F exposure activated mitochondrial-mediated apoptosis in the liver. However, the addition of 1% CaCO3 to the diet significantly increased the expression of anti-apoptotic gene Bcl-2 (P < 0.01), inhibited the activation of the mitochondrial apoptosis pathway, and reduced mitochondrial damage. In summary, supplementing 1% CaCO3 in the diet can alleviate the NaF-induced liver cell damage through the mitochondrial apoptosis pathway.


Asunto(s)
Calcio en la Dieta , Enfermedad Hepática Crónica Inducida por Sustancias y Drogas , Animales , Apoptosis , Enfermedad Hepática Crónica Inducida por Sustancias y Drogas/metabolismo , Fluoruros/metabolismo , Flúor , Hígado/metabolismo , Estrés Oxidativo , Ratas , Ratas Sprague-Dawley
5.
Zhongguo Dang Dai Er Ke Za Zhi ; 23(11): 1174-1183, 2021 Nov 15.
Artículo en Inglés, Chino | MEDLINE | ID: mdl-34753551

RESUMEN

OBJECTIVES: To study the effect of high-fat diet for maternal Sprague-Dawley rats at different stages on glucose and lipid metabolism in offspring and related mechanisms. METHODS: According to the diet before pregnancy and during pregnancy and lactation, maternal rats were randomly divided into four groups (n=9 each): CC (control diet before pregnancy and during pregnancy and lactation), HC (high-fat diet before pregnancy and control diet during pregnancy and lactation), CH (control diet before pregnancy and high-fat diet during pregnancy and lactation), and HH (high-fat diet before pregnancy and during pregnancy and lactation), and all offspring rats were given control diet after weaning. The body weight of maternal rats was recorded before and during pregnancy. Male offspring rats were selected from each group at the juvenile stage (3-week old) and the adult stage (12-week old) to measure the levels of fasting blood glucose (FBG) and fasting insulin (FINS) and the levels of triglyceride (TG) and total cholesterol (TC) in the liver. Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) index was calculated, and the area under the curve (AUC) was calculated for glucose tolerance test (GTT) and insulin tolerance test (ITT). Lipid deposition in the liver was observed, and the mRNA and protein expression levels of the key genes in glucose and lipid metabolism (IR, IRS, and AKT), FASN, SREBP1c, and PPARα in the liver were also measured. RESULTS: Compared with the control diet groups (CC and CH groups), the groups with high-fat diet before pregnancy (HC and HH groups) had a significant increase in body weight (P<0.001). Compared with the CC group, the HC, CH, and HH groups had a significantly greater increase in body weight (P<0.001). Compared with the CC group, the HC, CH, and HH groups had significant increases in body weight, the levels of TG and TC in the liver, and the mRNA and protein expression levels of FASN, SREBP1c, and PPARα in the offspring rats at week 3 after birth (P<0.05), as well as a significant increase in lipid deposition in the liver, with the most significant increase of the parameters in the HH group. Compared with the CC group, the HH group had significant increases in the levels of FBG and FINS, HOMA-IR index, GTT-AUC, ITT-AUC, and the protein expression level of p-IRS in the liver and significant reductions in the mRNA and protein expression levels of IR and IRS in the liver in the offspring rats at week 3 after birth (P<0.05). Compared with the CC group, the HC, CH, and HH groups had significant increases in body weight, the levels of FBG and FINS, HOMA-IR index, GTT-AUC, ITT-AUC, the levels of TG and TC in the liver, protein expression level of p-IRS in the liver, and the mRNA and protein expression levels of FASN, SREBP1c, and PPARα in the offspring rats at week 12 after birth (P<0.05), as well as a significant increase in lipid deposition in the liver, with the most increase of the parameters in the HH group. Compared with the CC group, the HC, CH, and HH groups had significant reductions in the mRNA expression levels of IR, IRS, and AKT and the protein expression levels of IR, IRS, and p-AKT in the offspring rats at week 12 after birth (P<0.05). There were no significant differences in the levels of glucose and lipid metabolism between the HC and CH groups at various stages (P>0.05). CONCLUSIONS: High-fat diet for rats at different stages before and after pregnancy has different effects on glucose and lipid metabolism of offspring rats, and high-fat diet before pregnancy and during pregnancy and lactation has the greatest effect. The effect of high-fat diet on glucose and lipid metabolism of offspring rats is considered associated with the changes in the expression of genes involved in glucose and lipid metabolism.


Asunto(s)
Dieta Alta en Grasa , Glucosa , Resistencia a la Insulina , Metabolismo de los Lípidos , Animales , Peso Corporal , Dieta Alta en Grasa/efectos adversos , Femenino , Glucosa/metabolismo , Insulina , Hígado/metabolismo , Masculino , Embarazo , Ratas , Ratas Sprague-Dawley
6.
Zhongguo Zhen Jiu ; 41(11): 1249-55, 2021 Nov 12.
Artículo en Chino | MEDLINE | ID: mdl-34762379

RESUMEN

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Neiguan" (PC 6) on cardiac function and cardiomyocyte apoptosis in rats with acute myocardial ischemia (AMI), and to explore the correlation between myocardial protective effect of EA and inflammatory factors i.e. interleukin-1ß (IL-1ß) and interleukin-17 (IL-17) of "Neiguan" (PC 6) area. METHODS: A total of 40 male SD rats with normal ultrasonic cardiogram were randomized into a sham-operation group, a sham-operation plus EA group, a model group and an EA group, 10 rats in each group. The AMI model was established by ligating the left anterior descending (LAD) branch of the coronary artery in the model group and the EA group, while the threading without ligating was adopted in the sham-operation group and the sham-operation plus EA group. In the sham-operation plus EA group and the EA group, EA at bilateral "Neiguan" (PC 6) was applied, with disperse-dense wave, 2 Hz/100 Hz in frequency and 2 mA in density, once a day, 20 min a time for 3 days. The cardiac ejection fraction (EF) and fractional shortening (FS) were measured by ultrasonic cardiogram to evaluate the cardiac function, the cardiomyocyte apoptosis was detected by TUNEL staining, the infiltration of inflammatory factors of "Neiguan" (PC 6) area was observed by H.E. staining, the expression of inflammatory factors IL-1ß and IL-17 of "Neiguan" (PC 6) area was detected by immunofluorescence staining. RESULTS: Compared with the sham-operation group, EF and FS were decreased (P<0.001), the average optical density of cardiomyocyte apoptosis was increased (P<0.001), the infiltration of inflammatory factors was obvious in skin dermis of "Neiguan" (PC 6) area in the model group; the positive expression of IL-1ß and IL-17 of "Neiguan" (PC 6) area was increased in the model group and the sham-operation plus EA group (P<0.001, P<0.01). Compared with the model group, EF and FS were increased (P<0.01), the average optical density of cardiomyocyte apoptosis was decreased (P<0.01), the infiltration of inflammatory factors was aggravating in skin dermis of "Neiguan" (PC 6) area, and the positive expression of IL-1ß and IL-17 of "Neiguan" (PC 6) area was increased in the EA group (P<0.001, P<0.01). CONCLUSION: Electroacupuncture at "Neiguan" (PC 6) can improve the cardiac function and reduce the apoptosis of cardiomyocyte in rats with acute myocardial ischemia, its mechanism may be related to the regulation of the inflammatory factors of "Neiguan" (PC 6) area.


Asunto(s)
Electroacupuntura , Isquemia Miocárdica , Puntos de Acupuntura , Animales , Masculino , Isquemia Miocárdica/terapia , Miocardio , Ratas , Ratas Sprague-Dawley
7.
Ann Palliat Med ; 10(10): 10756-10767, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-34763437

RESUMEN

BACKGROUND: Myocardial infarction (MI) is one of the most common cardiovascular diseases, inducing severe myocardial injury and leading to high mortality. Bromodomain-containing protein 7 (BRD7), a member of bromodomain-containing protein family, is involved in multiple cellular processes, such as cell cycle, transcriptional regulation, and chromatin remodeling, but the functions of BRD7 in regulating MI-associated myocardial injury are still obscure. In this work, we investigated the effect of BRD7 on MI-induced myocardial injury in vitro and in vivo. METHODS: The MI model was established by ligating the left anterior descending coronary artery (LAD) of rats which were then injected with BRD7 short hairpin RNA (shRNA). The rat H9C2 cardiomyocytes were treated with hypoxia and injected with BRD7 shRNA. The expression of BRD7 in MI rat model, and hypoxia-treated H9C2 cells was detected by quantitative polymerase chain reaction (qPCR), western blot, and immunohistochemical staining. The effect of BRD7 was analyzed using western blot, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, echocardiography, and flow cytometry analysis. The expressions of Wnt/ß-catenin signaling relative proteins were determined by western blot. RESULTS: Significantly, BRD7 was highly expressed in MI patients, MI rat models, and hypoxia treated rat H9C2 cardiomyocytes. Echocardiography analysis demonstrated that the left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) were repressed in the MI rats relative to sham group rats, while the silencing of BRD7 rescued the dysfunction in the model. We also found that BRD7 silencing reduced cardiomyocyte apoptosis in both MI rats and H9C2 cells under the treatment of hypoxia. BRD7 silencing inhibited the activation of Wnt/ß-catenin signaling in H9C2 cells under the treatment of hypoxia. Moreover, Wnt agonist BML294 reversed the anti-apoptosis effect of BRD7 silencing in hypoxia-induced H9C2 cells. CONCLUSIONS: Collectively, we concluded that BRD7 contributed to MI-induced myocardial injury through activating Wnt/ß-catenin signaling. Targeting BRD7 may become a promising therapeutic strategy for the treatment of MI-induced myocardial injury.


Asunto(s)
MicroARNs , Infarto del Miocardio , Animales , Proteínas Cromosómicas no Histona , Humanos , Infarto del Miocardio/genética , Ratas , Ratas Sprague-Dawley , Volumen Sistólico , Función Ventricular Izquierda , Vía de Señalización Wnt , beta Catenina/genética , beta Catenina/metabolismo
8.
Investig Clin Urol ; 62(6): 690-696, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34729968

RESUMEN

PURPOSE: The purpose of this study was to investigate the effect of aging on bladder function and caveolin protein expression in rat urothelium. MATERIALS AND METHODS: Female Sprague-Dawley rats were divided into the following two groups: young age control group (12 weeks) and old-aged group of rats (80 weeks). Urodynamic measurements were taken to compare the contraction interval and the contraction pressure between the two groups. The expression and cellular localization of caveolin 1 and 2 in the urothelium of the rat urinary bladder were determined by Western blot and immunofluorescence microscopy. RESULTS: In cystometrograms, the contraction interval (min) was significantly shorter in the old-aged group (3.7±0.5 min) than in the young age control group (6.2±0.8 min). Also, the average contraction pressure (mmHg) was lower in the old-aged group (8.4±0.6 mmHg) than in the young age control group (13.2±1.3 mmHg). Caveolin 1 and 2 were expressed in the subepithelial area in the urothelium. The protein expression of both caveolin 1 and 2 was significantly lower in the old-aged group than in the young age control group. CONCLUSIONS: Aging caused a significant change in the expression of caveolin 1 and 2 in the urothelium of the rat urinary bladder. These findings suggest that these molecules might have specific roles in the functional change of the urinary bladder that occurs in association with aging.


Asunto(s)
Vejiga Urinaria Hiperactiva , Urotelio , Envejecimiento , Animales , Caveolina 1 , Femenino , Ratas , Ratas Sprague-Dawley
9.
Nan Fang Yi Ke Da Xue Xue Bao ; 41(10): 1448-1455, 2021 Oct 20.
Artículo en Chino | MEDLINE | ID: mdl-34755659

RESUMEN

OBJECTIVE: To evaluate the efficacy and safety of attenuated Herpes simplex virus 1 (HSV-1) vector expressing oncomodulin (OCM) for treatment of mechanical optic nerve injury in rats. METHODS: The proliferation characteristics and OCM expression of the recombinant HSV-1 vector (1716-OCM) was assessed in cultured Vero cells. Twelve-week-old SD rats were randomly divided into control group, 1716-OCM injection group and wild-type virus corneal infection group, and at 7, 14, 30 and 60 days post-infection (3 rats in each group at each time point), the expressions of OCM and HSV-1 structural protein gB in the retina and the hypothalamus of the rats were detected using immunofluorescence assay. Another 20 rats were randomized into sham operation group, PBS treatment group, 1716-OCM infection group and 1716-OCM infection with cAMP sensitization group (n=5), and in the latter 3 groups, rat models of optic nerve injury models were established followed by intravitreal injection of PBS, 1716-OCM or cAMP as indicated. At 45 days after the treatments, the rats were examined for visual electrophysiological function using FVEP method, and the number of retinal ganglion cells (RGCs) and the expression of myelin basic protein in the optic nerve were detected using immunofluorescence assay. RESULTS: The recombinant 1716-OCM vector was capable of mediating effective expression of OCM in Vero cells in vitro, but its proliferation rate was much lower than that of the wild-type virus. In SD rats, the recombinant virus could mediate the expression of OCM in the RGC layer and choroid layer of the eyes without inducing significant structural damage of the eyes as compared with the wild-type virus. In rat models of optic nerve injury, 1716-OCM combined with cAMP significantly promoted the survival of retinal RGCs (P= 0.007) and inhibited demyelination of the optic nerve (P=0.03) as compared with the mock treatment. FVEP analysis showed that 1716-OCM combined with cAMP significantly promoted the recovery of the peak amplitude of ΔN1-P1 in the rats (P < 0.0001). CONCLUSION: Attenuated recombinant 1716-OCM vector can mediate OCM expression in the retina of rats, and in rat models of mechanical optic nerve injury, intravitreal injection of 1716-OCM combined with cAMP can effectively alleviate optic nerve injuries.


Asunto(s)
Herpesvirus Humano 1 , Traumatismos del Nervio Óptico , Animales , Proteínas de Unión al Calcio , Chlorocebus aethiops , Modelos Animales de Enfermedad , Herpesvirus Humano 1/genética , Ratas , Ratas Sprague-Dawley , Células Vero
10.
Nan Fang Yi Ke Da Xue Xue Bao ; 41(10): 1464-1472, 2021 Oct 20.
Artículo en Chino | MEDLINE | ID: mdl-34755661

RESUMEN

OBJECTIVE: To explore the association of the Notch1/Jagged1 pathway with the homing of mesenchymal stem cells (BMSCs) to regulate Th1/Th2 drift in asthma. METHODS: Twenty SD rats were randomly divided into normal control group, model group, BMSC transplantation group, and BMSC+Notch inhibitor group. Ovalbumin sensitization was used to establish rat models of asthma, and BMSCs were transplanted via the tail vein. The pathology of the lung tissue was examined with HE staining, and the contents of interleukin (IL)-5, IL-13, and interferon-γ (IFN-γ) in lung tissue homogenate were determined with enzyme-linked immunosorbent assay. The expressions of Notch1 and Jagged1 mRNA were detected with RT-PCR, and CXCR4 expression in the bronchial epithelial cells was examined using immunofluorescence staining; Western blotting was used to detect the protein expressions of T-bet, GATA-3, Notch1, and Jagged1 in the lung tissue. RESULTS: Compared with those in the control group, the expressions of IFN-γ and T-bet proteins decreased significantly and the pulmonary expressions of IL-5, IL-13, and GATA-3 proteins as well as Notch1 and Jagged1 mRNA and protein expressions all increased significantly in the model group (P < 0.05 or 0.01). Compared with those in the model group, CXCR4, IFN-γ, and T-bet protein expressions in BMSC group and BMSCs+Notch inhibitor group all increased significantly, and Notch1 and Jagged1 protein expressions in BMSCs group and IL-5, IL-13, Notch1, and Jagged1 mRNA and protein expressions in BMSCs + Notch inhibitor group all decreased significantly (P < 0.05 or 0.01). The expressions of CXCR4 and IFN-γ were significantly higher and the expressions of IL-13 and Notch1 mRNA were significantly lower in BMSCs+Notch inhibitor group than in BMSC group (P < 0.05). CONCLUSION: In asthmatic rats, the homing of the BMSCs to the lung tissue has a regulatory effect on Th1/Th2 drift, and the Notch1/Jagged1 pathway may participate in the homing of the BMSCs.


Asunto(s)
Asma , Células Madre Mesenquimatosas , Animales , Proteína Jagged-1/genética , Pulmón/metabolismo , Células Madre Mesenquimatosas/metabolismo , Ratas , Ratas Sprague-Dawley , Receptor Notch1/genética , Receptor Notch1/metabolismo
11.
Nan Fang Yi Ke Da Xue Xue Bao ; 41(10): 1519-1526, 2021 Oct 20.
Artículo en Chino | MEDLINE | ID: mdl-34755667

RESUMEN

OBJECTIVE: To explore the role of cell cycle checkpoint kinase 1/2 (CHK1/2) in mediating the inhibitory effect of oxymatrine (OMT) against renal inflammation and fibrosis in diabetic rats. METHODS: SD rats were randomly divided into normal control group, diabetes model group (DM) and OMT treatment group (n=6). HE and Masson staining were used to observe histopathological changes of the renal tissue, and the expressions of CHK1, CHK2, p-CHK1 and p-CHK2 were localized by immunohistochemical staining. The contents of interleukin-6 (IL-6) and IL-1ß in the renal tissue were detected using ELISA, and the expression levels of CHK1, CHK2, p-CHK1, p-CHK2, type Ⅲ collagen (Col-Ⅲ), type Ⅳ collagen (Col-Ⅳ), and fibronectin (FN) were determined using Western blotting. The changes in the expressions of CHK1, CHK2, p-CHK1, p-CHK2, Col-Ⅲ, Col-Ⅳ and FN proteins were also examined with Western blotting in NRK-52E cells in response to high glucose exposure, OMT treatment and siRNA-mediated CHK1/2 knockdown. RESULTS: In diabetic rats, OMT treatment significantly decreased the levels of blood glucose, serum creatinine and 24 h urinary protein (P < 0.05) and obviously improved inflammatory cell infiltration and fibrosis phenotype in the renal tissue (P < 0.05). CHK1 and CHK2 were mainly expressed in the cytoplasm and nuclei of renal tubule cells, and their phosphorylation levels were significantly higher in DM group than in the control group and OMT group. OMT treatment significantly decreased the protein expression levels of p-CHK1, p-CHK2, Col-Ⅲ, Col-Ⅳ and FN in the renal tissue of diabetic rats and in NRK-52E cells exposed to high glucose (P < 0.05). In NRK-52E cells, CHK1/2 knockdown resulted in significant reduction of the protein expressions of p-CHK1/2, Col-Ⅲ, Col-Ⅳ and FN (P < 0.05). CONCLUSION: The inhibitory effects of OMT against renal inflammation and fibrosis in diabetic rats are mediated probably by lowered phosphorylation levels of CHK1 and CHK2, which result in reduced release of the downstream inflammatory mediators and decreased secretion and deposition of extracellular matrix.


Asunto(s)
Diabetes Mellitus Experimental , Alcaloides , Animales , Diabetes Mellitus Experimental/complicaciones , Fibrosis , Inflamación/tratamiento farmacológico , Fosforilación , Quinolizinas , Ratas , Ratas Sprague-Dawley
12.
Nan Fang Yi Ke Da Xue Xue Bao ; 41(10): 1547-1553, 2021 Oct 20.
Artículo en Chino | MEDLINE | ID: mdl-34755671

RESUMEN

OBJECTIVE: To detect the binding of Mage-D1 with activated p75NTR and explore their role in regulating mineralization of ectomesenchymal stem cells (EMSCs). METHODS: EMSCs were isolated from the tooth germs of embryonic SD rats (19.5 days of gestation) by tissue explant culture and were identified for surface markers using flow cytometry. The cultured cells were divided into blank control group, 100 ng/mL nerve growth factor (NGF) stimulation group, and lentivirus-mediated Mage-D1 interference (SH-Mage-D1) group. Proximity ligation assay was used to detect the binding of Mage-D1 with activated p75NTR in the EMSCs, and the binding strength was compared among the 3 groups. Alizarin red staining and ALP staining were used to observe mineralization of the induced cells. The expressions of ALP, Runx2, OCN, BSP, OPN, Msx1 and Dlx1 at both the mRNA and protein levels were detected using RT-PCR and Western blotting. RESULTS: The isolated EMSCs expressed high levels of cell surface markers CD44, CD90, CD29, CD146, and CD105 with a low expression of CD45. The results of proximity ligation assay showed that the binding of Mage-D1 with activated p75NTR in the cells increased over time, and the binding strength was significantly greater in NFG-treated cells than in the cells in the other two groups (P < 0.05). Alizarin red staining and ALP staining of the induced cells showed that the changes in the mineralization nodules were consistent with those of ALP activity. The cells treated with 100 ng/mL NGF exhibited significantly increased expressions of ALP, Runx2, OCN, BSP, OPN, Col1, Msx1 and Dlx1 as compared with the cells in the other two groups (P < 0.05). CONCLUSION: Mage-D1 directly binds to activated p75NTR in embryonic rat EMSCs to positively regulate the mineralization of the EMSCs.


Asunto(s)
Células Madre Mesenquimatosas , Animales , Diferenciación Celular , Células Cultivadas , Lentivirus , Osteogénesis , Ratas , Ratas Sprague-Dawley
13.
BMC Musculoskelet Disord ; 22(1): 944, 2021 Nov 11.
Artículo en Inglés | MEDLINE | ID: mdl-34763682

RESUMEN

BACKGROUND: Poor osseointegration is the key reason for implant failure after arthroplasty,whether under osteoporotic or normal bone conditions. To date, osseointegration remains a major challenge. Recent studies have shown that deferoxamine (DFO) can accelerate osteogenesis by activating the hypoxia signaling pathway. The purpose of this study was to test the following hypothesis: after knee replacement, intra-articular injection of DFO will promote osteogenesis and osseointegration with a 3D printed titanium prosthesis in the bones of osteoporotic rats. MATERIALS AND METHODS: Ninety female Sprague-Dawley rats were used for the experiment. Ten rats were used to confirm the successful establishment of the osteoporosis model: five rats in the sham operation group and five rats in the ovariectomy group. After ovariectomy and knee arthroplasty were performed, the remaining 80 rats were randomly divided into DFO and control groups (n = 40 per group). The two groups were treated by intraarticular injection of DFO and saline respectively. After 2 weeks, polymerase chain reaction (PCR) and immunohistochemistry were used to evaluate the levels of HIF-1a, VEGF, and CD31. HIF-1a and VEGF have been shown to promote angiogenesis and bone regeneration, and CD31 is an important marker of angiogenesis. After 12 weeks, the specimens were examined by micro-computed tomography (micro-CT), biomechanics, and histopathology to evaluate osteogenesis and osseointegration. RESULTS: The results of PCR showed that the mRNA levels of VEGF and CD31 in the DFO group were significantly higher than those in the control group. The immunohistochemistry results indicated that positive cell expression of HIF-1a, VEGF, and CD31 in the DFO group was also higher. Compared with the control group, the micro-CT parameters of BMD, BV/TV, TB. N, and TB. Th were significantly higher. The maximal pull-out force and the bone-to-implant contact value were also higher. CONCLUSIONS: The local administration of DFO, which is used to activate the HIF-1a signaling pathway, can promote osteogenesis and osseointegration with a prosthesis in osteoporotic bone.


Asunto(s)
Miembros Artificiales , Titanio , Animales , Femenino , Humanos , Hipoxia , Oseointegración , Ovariectomía , Ratas , Ratas Sprague-Dawley , Microtomografía por Rayos X
14.
Int J Mol Sci ; 22(21)2021 Oct 21.
Artículo en Inglés | MEDLINE | ID: mdl-34768820

RESUMEN

Disseminated intravascular coagulation (DIC) is a severe condition characterized by the systemic formation of microthrombi complicated with bleeding tendency and organ dysfunction. In the last years, it represents one of the most frequent consequences of coronavirus disease 2019 (COVID-19). The pathogenesis of DIC is complex, with cross-talk between the coagulant and inflammatory pathways. The objective of this study is to investigate the anti-inflammatory action of ultramicronized palmitoylethanolamide (um-PEA) in a lipopolysaccharide (LPS)-induced DIC model in rats. Experimental DIC was induced by continual infusion of LPS (30 mg/kg) for 4 h through the tail vein. Um-PEA (30 mg/kg) was given orally 30 min before and 1 h after the start of intravenous infusion of LPS. Results showed that um-PEA reduced alteration of coagulation markers, as well as proinflammatory cytokine release in plasma and lung samples, induced by LPS infusion. Furthermore, um-PEA also has the effect of preventing the formation of fibrin deposition and lung damage. Moreover, um-PEA was able to reduce the number of mast cells (MCs) and the release of its serine proteases, which are also necessary for SARS-CoV-2 infection. These results suggest that um-PEA could be considered as a potential therapeutic approach in the management of DIC and in clinical implications associated to coagulopathy and lung dysfunction, such as COVID-19.


Asunto(s)
Amidas/uso terapéutico , Trastornos de la Coagulación Sanguínea/tratamiento farmacológico , Coagulación Intravascular Diseminada/tratamiento farmacológico , Etanolaminas/uso terapéutico , Ácidos Palmíticos/uso terapéutico , Sepsis/complicaciones , Amidas/química , Amidas/farmacología , Animales , Trastornos de la Coagulación Sanguínea/etiología , COVID-19/patología , COVID-19/virología , Citocinas/sangre , Citocinas/metabolismo , Modelos Animales de Enfermedad , Coagulación Intravascular Diseminada/etiología , Etanolaminas/química , Etanolaminas/farmacología , Productos de Degradación de Fibrina-Fibrinógeno/metabolismo , Lipopolisacáridos/toxicidad , Pulmón/metabolismo , Pulmón/patología , Masculino , Mastocitos/citología , Mastocitos/efectos de los fármacos , Mastocitos/metabolismo , Ácidos Palmíticos/química , Ácidos Palmíticos/farmacología , Tiempo de Tromboplastina Parcial , Tiempo de Protrombina , Ratas , Ratas Sprague-Dawley , SARS-CoV-2/aislamiento & purificación , Sepsis/patología , Serina Proteasas/metabolismo
15.
J Int Med Res ; 49(11): 3000605211053549, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34743632

RESUMEN

OBJECTIVE: Inflammation is an important factor in the pathological process of cerebral ischemia. Artesunate exhibits a broad range of anti-inflammatory properties in many diseases. We investigated the potential protective effect of artesunate against cerebral ischemia and the related mechanisms. METHODS: Mice were divided into distal middle cerebral artery occlusion (dMCAO), sham, low dose, and high dose groups and subjected to dMCAO, except for the sham group. The low and high dose groups were administered artesunate (15 and 30 mg/kg), and the neuroprotective effects were analyzed by evaluating infarct volumes and neurological deficits. Microglial activation and neutrophil infiltration were evaluated by immunofluorescence, immunohistochemical staining, and western blotting. Inflammatory mediators were measured by enzyme-linked immunosorbent assays. Nuclear factor (NF)-κB nuclear translocation was detected by immunofluorescence and western blotting. RESULTS: Compared with the dMCAO group, artesunate significantly improved neurological deficit scores and infarct volumes and ameliorated inflammation by reducing neutrophil infiltration, suppressing microglial activation, and downregulating tumor necrosis factor-α and interleukin-1ß expression. Furthermore, artesunate inhibited nuclear translocation of NF-κB and inhibitor protein α proteolysis. CONCLUSIONS: Artesunate protected against inflammatory injury by reducing neutrophil infiltration and microglial activation, suppressing inflammatory cytokines, and inhibiting the NF-κB pathway. Therefore, artesunate is a potential ischemic stroke treatment.


Asunto(s)
Isquemia Encefálica , FN-kappa B , Animales , Artesunato , Isquemia Encefálica/tratamiento farmacológico , Infarto de la Arteria Cerebral Media/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Ratones , FN-kappa B/genética , FN-kappa B/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal
16.
J Agric Food Chem ; 69(45): 13618-13627, 2021 Nov 17.
Artículo en Inglés | MEDLINE | ID: mdl-34735150

RESUMEN

Using Sprague-Dawley rats and rat PC12 cells treated with sodium fluoride (NaF), we investigated the effects of SIK2-CRTC1 signaling on the neurobehavioral toxicity induced by fluoride. The in vivo results demonstrated that NaF treatment induced anxiety- and depression-like behaviors in juvenile rats, resulting in histological and ultrastructural abnormalities in the rat hippocampus and medial prefrontal cortex. Moreover, NaF exposure induced neuronal loss and excessive apoptosis. We also found that NaF elevated the expression of SIK2 and reduced the expression of CRTC1, brain-derived neurotrophic factor (BDNF), and VGF. The in vitro results showed that NaF suppressed cell viability, induced SIK2-CRTC1 signaling dysfunction, and caused excessive apoptosis in PC12 cells. Notably, targeted knockout of SIK2 with SIK2-siRNA or blocking of SIK2-CRTC1 signaling with 7,8-dihydroxyflavone (7,8-DHF) (as well as venlafaxine) can reduce apoptosis and increase cell viability in vitro. These findings suggest that neuronal death resulting from abnormal SIK2-CRTC1 signaling contributes to neurobehavioral toxicity induced by fluoride.


Asunto(s)
Depresión , Fluoruros , Animales , Ansiedad/inducido químicamente , Ansiedad/genética , Apoptosis , Depresión/inducido químicamente , Depresión/genética , Proteínas Serina-Treonina Quinasas/genética , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Factores de Transcripción
17.
Acta Cir Bras ; 36(9): e360906, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34755766

RESUMEN

PURPOSE: To evaluate the effect of hyperbaric oxygenation (HBO) on angiogenesis in random rat skin flaps, by immunoexpression of vascular endothelial growth factor A (VEGF-A). METHODS: Forty adult rats were divided into four groups: GE) epilated; GE/HBO) epilated subjected to HBO; GER) epilated submitted to dorsal skin flap; GER/HBO) epilated subjected to dorsal skin flap + HBO. HBO was performed with rats inside a chamber under atmosphere close to 100% oxygen and pressure of 2.4 absolute atmospheres, 2h per day during seven consecutive days. GE and GER groups were placed in the hyperbaric chamber without HBO. Then, under anesthesia, skin flaps were removed and separated into three portions relative to pedicle fixation. The samples were fixed in formalin and processed for paraffin embedding. Histological sections were submitted to immunohistochemistry for VEGF-A detection. The number of immunostained-blood vessels were counted under light microscopy. RESULTS: GE and GE/HBO groups showed normal and similar skin morphology in the three flap portions. A fibrin-leukocyte crust, along with denatured collagen and intense leukocyte infiltrate, was mainly observed in the dermis of the medial and distal flap portions of GER group. Meanwhile, the GER/HBO group presented more regions with intact collagen and small areas of leukocyte infiltrate in the three flap regions. VEGF-A-immunostained blood vessels were largely seen in all regions of GE and GE/HBO groups, whereas no significant differences were found between these groups. A decrease in vascularization was noticed in GER and GER/HBO groups, which was more evident in the most distal portion of the flaps. However, the number of VEGF-A-immunostained blood vessels in GER/HBO group was significantly higher when compared to GER group. CONCLUSIONS: Hyperbaric oxygenation was associated with increased angiogenesis and improved viability of rat skin flaps.


Asunto(s)
Oxigenación Hiperbárica , Animales , Ratas , Ratas Sprague-Dawley , Trasplante de Piel , Colgajos Quirúrgicos , Factor A de Crecimiento Endotelial Vascular
18.
BMC Musculoskelet Disord ; 22(1): 949, 2021 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-34781961

RESUMEN

INTRODUCTION: Tendon diseases and injuries are a serious problem for the aged population, often leading to pain, disability and a significant decline in quality of life. The purpose of this study was to determine the influence of aging on biochemistry and histology during tendon healing and to provide a new strategy for improving tendon healing. METHOD: A total of 24 Sprague-Dawley rats were equally divided into a young and an aged group. A rat patellar tendon defect model was used in this study. Tendon samples were collected at weeks 2 and 4, and hematoxylin-eosin, alcian blue and immunofluorescence staining were performed for histological analysis. Meanwhile, reverse transcription-polymerase chain reaction (RT-PCR) and western blot were performed to evaluate the biochemical changes. RESULTS: The histological scores in aged rats were significantly lower than those in young rats. At the protein level, collagen synthesis-related markers Col-3, Matrix metalloproteinase-1 and Metallopeptidase Inhibitor 1(TIMP-1) were decreased at week 4 in aged rats compared with those of young rats. Though there was a decrease in the expression of the chondrogenic marker aggrecan at the protein level in aged tendon, the Micro-CT results from weeks 4 samples showed no significant difference(p>0.05) on the ectopic ossification between groups. Moreover, we found more adipocytes accumulated in the aged tendon defect with the Oil Red O staining and at the gene and protein levels the markers related to adipogenic differentiation. CONCLUSIONS: Our findings indicate that tendon healing is impaired in aged rats and is characterized by a significantly lower histological score, decreased collagen synthesis and more adipocyte accumulation in patellar tendon after repair.


Asunto(s)
Calidad de Vida , Cicatrización de Heridas , Envejecimiento , Animales , Ratas , Ratas Sprague-Dawley , Tendones
19.
J Int Med Res ; 49(11): 3000605211055059, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34772311

RESUMEN

OBJECTIVE: To investigate the effect of liraglutide on the browning of white fat and the suppression of obesity via regulating microRNA (miR)-27b in vivo and in vitro. METHODS: Sprague-Dawley rats were fed a high-fat (HF) diet and 3T3-L1 pre-adipocytes were differentiated into mature white adipocytes. Rats and mature adipocytes were then treated with different doses of liraglutide. The mRNA and protein levels of browning-associated proteins, including uncoupling protein 1 (UCP1), PR domain containing 16 (PRDM16), CCAAT enhancer binding protein ß (CEBPß), cell death-inducing DFFA-like effector A (CIDEA) and peroxisome proliferator-activated receptor-γ-coactivator 1α (PGC-1α), were detected using quantitative real-time polymerase chain reaction and Western blotting. RESULTS: Liraglutide decreased body weight and reduced the levels of blood glucose, triglyceride and low-density lipoprotein cholesterol in HF diet-fed rats. Liraglutide increased the levels of UCP1, PRDM16, CEBPß, CIDEA and PGC-1α in vivo and vitro. The levels of miR-27b were upregulated in HF diet-fed rats, whereas liraglutide reduced the levels of miR-27b. In vitro, overexpression of miR-27b decreased the mRNA and protein levels of UCP1, PRDM16, CEBPß, CIDEA and PGC-1α. Transfection with the miR-27b mimics attenuated the effect of liraglutide on the browning of white adipocytes. CONCLUSION: Liraglutide induced browning of white adipose through regulation of miR-27b.


Asunto(s)
Liraglutida , MicroARNs , Tejido Adiposo Pardo , Tejido Adiposo Blanco , Animales , Dieta Alta en Grasa/efectos adversos , Liraglutida/farmacología , MicroARNs/genética , Obesidad/tratamiento farmacológico , Obesidad/genética , Ratas , Ratas Sprague-Dawley
20.
An Acad Bras Cienc ; 94(1): e20210314, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34787291

RESUMEN

Excessive exercise leads to myocardial injury or even sudden exercise death. For the vast sports population, appropriate physiological state is a necessary condition for exercise. The present study aims to investigate the cardioprotective effects and potent mechanism of astragalus polysaccharide (APS) treatment against the exercise-induced myocardial injury via in vitro cell-based assay and in vivo model rat. Efficacies of APS incubation on the inflammatory response and oxidative stress induced by LPS were both explored in H9c2 cells by using CCK-8 and western blotting method, respectively. Normal SD rats were randomly divided into saline-treated overexercise rat group, and APS-treated overexercise rat groups with three doses. Then long-term swimming training load cycle (8 week) were performed on these rats. Finally, the changes on body weight, myocardial morphological and injury indicators, as well as the inflammation-related proteins in overexercise-induced model rats were all assessed. Three concentrations of APS all significantly increased cell viability, and decreased the apoptosis of cardiomyocytes in LPS-treated H9c2 cells. Moreover, chronic treatment of APS at all three doses also could obviously decreased myocardial injury-related indicators. Furthermore, the histopathologic examination exhibited that the APS successfully attenuated the changes of myocardial tissues, reduced the lipid accumulation and the protein levels of IL-1ß, TNF-α and NF-κB. Furthermore, the APS could activate the AMPK signaling pathway, enhance the autophagy and suppress the production of ROS. On conclusions, APS exerted the protective efficacies on overexercise-induced myocardial injury by activating the AMPK signaling pathway to increase autophagy and suppress the inflammation response, oxidative stress, apoptosis of myocardial cells.


Asunto(s)
Proteínas Quinasas Activadas por AMP , Transducción de Señal , Proteínas Quinasas Activadas por AMP/metabolismo , Animales , Apoptosis , Inflamación/tratamiento farmacológico , Estrés Oxidativo , Polisacáridos/farmacología , Ratas , Ratas Sprague-Dawley
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