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ObjectiveTo investigate the mechanism of Xiaonang Tiaojing decoction（XNTJD）in improving polycystic ovary syndrome with insulin resistance（PCOS-IR）model rats based on anti-Müllerian hormone（AMH）/AMH type Ⅱ receptor（AMHRⅡ）signaling pathway. MethodForty-eight adult female SD rats were randomly divided into the blank group， model group， XNTJD group（11.4 g·kg-1·d-1） and Diane-35 group（0.21 g·kg-1·d-1）， PCOS-IR model was established by high-fat and high-sugar diet combined with letrozole in rats of all groups except the blank group， rats in the administration groups were given the corresponding dose of drugs by gavage for 15 days with an interval of 1 d every 4 d， normal saline of the same volume was given to the blank group and the model group. Estrous cycle was recorded daily during treatment. At the end of treatment， body weight and Lee's index were recorded， AMH， luteinizing hormone（LH）， LH/follicle stimulating hormone（FSH）， testosterone（T）and estradiol（E2）levels were measured by enzyme-linked immunosorbent assay（ELISA）， fasting plasma glucose（FPG）was measured by glucometer， fasting insulin（FINS） level was measured by radioimmunoassay（RIA）， and the insulin resistance index（HOMA-IR） and insulin sensitivity index（QUICKI）were calculated， triglyceride（TG）and total cholesterol（TC）levels were measured by automatic biochemical analyzer， hematoxylin-eosin（HE）staining was used to observe the morphological changes of the ovary， the levels of AMHRⅡ， bone morphogenetic protein-15（BMP-15）and Smad5 in ovarian tissues were detected by immunohistochemistry（IHC），Western blot was used to analyze the protein expression levels of AMHRⅡ， BMP-15 and Smad5. ResultCompared with the blank group， a large number of leukocytes were observed in the vaginal exfoliated cells of rats in the model group， mainly in diestrus， the levels of body weight， Lee's index， glucose-lipid metabolism indexes（FPG， FINS， HOMA-IR， TG， TC）， AMH and sex hormones（LH， LH/FSH， T）were significantly increased（P<0.01）， and QUICKI and E2 levels were significantly decreased（P<0.01）， there were more cystic bulges on the ovarian surface， more wet weight， more atretic and cystic dilated follicles in the ovarian tissues， and the thickness of granulosa cell layer was reduced without oocytes， the expression level of AMHRⅡ protein in ovarian tissues was significantly increased（P<0.01）， and the expression levels of BMP-15 and Smad5 proteins were significantly decreased（P<0.01）. Compared with the model group， the exfoliated cells in the vagina of rats treated with XNTJD group showed keratinocytes from the 5th to 6th day of treatment， and a stable estrous cycle gradually appeared， body weight， Lee's index， glucose-lipid metabolism indexes（FPG， FINS， HOMA-IR， TG， TC）， AMH and sex hormones（LH， LH/FSH， T）levels were significantly decreased（P<0.05， P<0.01）， QUICKI and E2 levels were significantly increased（P<0.01）， ovarian surface was smoother and lighter in wet weight， oocytes and mature follicles were observed in ovarian tissues， the thickness of granulosa cell layer increased and the morphology was intact， the expression levels of BMP-15 and Smad5 proteins were significantly increased（P<0.01）and the expression level of AMHRⅡ protein was significantly decreased（P<0.01）in ovarian tissues. ConclusionXNTJD may mediate the up-regulation of BMP-15 and Smad5 in ovarian tissues of PCOS-IR rats by down-regulating AMH/AMHRⅡ， thereby improving ovarian function， sex hormones and glucose-lipid metabolism levels in PCOS-IR rats.
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ObjectiveTo explore the effects and mechanisms of Kaiyu Zhongyutang on insulin resistance， glucose and lipid metabolism， psychological state， and embryo outcome in the infertile patients with polycystic ovary syndrome （PCOS） due to liver depression and kidney deficiency. MethodThe 126 infertile patients with PCOS due to liver depression and kidney deficiency who underwent in vitro fertilization-embryo transfer （IVF-ET） in the Department of Reproduction of the affiliated hospital of Nanjing university of Chinese medicine were randomly assigned into the observation （Kaiyu Zhongyutang + metformin） and control （metformin） groups. The two groups were compared in terms of traditional Chinese medicine （TCM） syndrome scores， body mass index （BMI）， glucose and lipid metabolism， homeostasis model assessment-insulin resistance （HOMA-IR）， interleukin （IL）-6， IL-8， C-reactive protein （CRP）， self-rating depression scale （SDS） score， self-rating anxiety scale （SAS） score， and clinical and laboratory scores of IVF after treatment. Result① After treatment， the observation group showed decreased scores of primary and secondary TCM syndromes and total TCM syndrome score （P < 0.05）， and the control group presented decreased scores of irritability and depression and total TCM syndrome score （P<0.05）. Compared with the control group， the observation group demonstrated reduced primary and secondary syndrome scores and total score after treatment （P<0.05）. ② After treatment， both groups showed decreased BMI， lowered levels of fasting insulin （FINS）， fasting plasma glucose （FPG）， cholesterol （TC）， triglyceride （TG）， low-density lipoprotein （LDL）， and HOMA-IR （P<0.05）， and elevated high-density lipoprotein （HDL） level （P<0.05）. Moreover， the observation group had lower BMI， FINS， FPG， TC， TG， LDL， and HOMA-IR and higher HDL than the control group （P<0.05）. ③ The treatment in both groups lowered the levels of IL-6， IL-8， and CRP and decreased SDS and SAS scores （P<0.05）. Moreover， the declines in the observation group were more obvious than those in the control group （P<0.05）. ④ Correlation analysis before treatment， IL-6， IL-8， and CRP had positive correlations with BMI， HOMA-IR， TC， TG， LDL， SDS， and SAS （P<0.05） and a negative correlation with HDL （P<0.05）. ⑤ The observation group showed reduced gonadotropin （Gn） using days and total Gn dose and higher two-pronuclear （2PN） fertilized oocytes， 2PN cleavage rate， normal fertilization rate， D3 transferable embryo number， D3 high-quality embryo number， high-quality embryo rate， and blastocyst formation rate than the control group （P<0.05）. ConclusionKaiyu Zhongyutang can treat PCOS patients by improving the emotional and reproductive functions and alleviating insulin resistance and glucose and lipid metabolism disorders. Moreover， it can reduce the Gn dose and Gn using days in the IVF process， improve the quality and maturity of eggs， increase the egg fertilization rate， enhance the potential of embryo development， and increase the rate of blastocyst formation by inhibiting inflammation.
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Type 2 diabetes mellitus （T2DM） is a heterogeneous disease with insulin deficiency and insulin resistance （IR） as the main etiology and is often accompanied by complications. Volatile oil is a volatile oily liquid extracted from natural plants， which has many pharmacological effects such as regulating Qi， relieving pain， inhibiting bacteria， and reducing inflammation. In recent years， there have been numerous reports on the treatment of T2DM by natural plant volatile oil and its effective components， which has become one of the new directions in the treatment of T2DM. With natural plant essential oil and its active components as the starting point， this paper comprehensively analyzed and summarized the material basis， mechanism， and signaling pathways of essential oil in the treatment of T2DM and its complications in China and abroad in recent years， and focused on the inhibitory effect of essential oil and its active components， such as carvacrol， paeonol， and β-caryophylene， on IR to improve T2DM by protecting pancreatic β-cells， inhibiting α-glucosidase activity， regulating the abundance and diversity of intestinal microbiota， and regulating glucose transporter protein type4 （GLUT4）， adenylate 5′-monophosphate-activated protein kinase （AMPK）， phosphatidylinositol-3 kinase （PI3K）/protein kinase B （Akt） signaling pathways to provide some references for the volatile oil intervention in T2DM and the development of new green antidiabetic drugs.
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This study investigated the mechanism of improving impaired glucose tolerance(IGT) of rats by Huanglian Wendan Decoction from the perspective of the skeletal muscle Nod-like receptor protein 3(NLRP3)/cysteinyl aspartate specific proteinase-1(caspase-1)/interleukin-1β(IL-1β), interleukin-18(IL-18) pathway. Healthy male SD rats were fed with the diet containing 45% fat for 20 weeks, accompanied by a high-temperature and high-humidity environment and an inactive lifestyle, for the establishment of the IGT rat model. The rats were divided into the blank control group, model control group, metformin hydrochloride group(positive drug group, 0.05 g·kg~(-1)·d~(-1)) and Huanglian Wendan Decoction group(7.8 g·kg~(-1)·d~(-1)). After continuous intragastric administration for 4 weeks, the obesity and glycemic indexes of all the rats were measured. The fasting serum insulin(FINS) level was determined by ELISA, with the insulin sensitivity index(ISI) and insulin resistance index(IRI) calculated. The mRNA and protein expression le-vels of nuclear factor kappaB(NF-κB), NLRP3, caspase-1, IL-1β and IL-18 in skeletal muscle tissue were detected by real-time polymerase chain reaction(PCR), Western blot and immunofluorescence. Compared with the blank control group, the model control group witnessed significantly increased mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1β and IL-18. As revealed by the comparison with the model control group, Huanglian Wendan Decoction could effectively regulate the obesity status, reduce body weight, correct the abnormal levels of 2-hour plasma glucose(2 hPG), insulin resistance index(IRI), insulin sensitivity index(ISI), and lower the mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1β and IL-18 in the skeletal muscle tissue of IGT rats. Combined with previous studies, the above results showed that the occurrence and development of IGT was closely related to inflammatory response and the classic pyroptosis pathway in skeletal muscle, such as NLRP3/caspase-1/IL-1β, IL-18. It is inferred that the mechanism of Huanglian Wendan Decoction was to alleviate insulin resistance(IR) and then reverse the course of IGT lies in the regulation of the abnormal insulin receptor signaling pathway based on the NLRP3 inflammasome pathway.
Subject(s)
Animals , Male , Rats , Caspase 1/genetics , Drugs, Chinese Herbal , Glucose Intolerance , Interleukin-18/genetics , Interleukin-1beta , Muscle, Skeletal , NF-kappa B/genetics , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , Rats, Sprague-DawleyABSTRACT
To investigate the mechanism of Huanglian Wendan Decoction in intervening impaired glucose tolerance( IGT) rat insulin resistance( IR) based on the pyroptosis pathway of liver cells. The IGT rat model was established by high-fat diet( 20 weeks) combined with high temperature,humidity environment and inactive lifestyle. The experiment was divided into blank group,model group,metformin hydrochloride group( positive group)( 0. 05 g·kg~(-1)·d~(-1)) and Huanglian Wendan Decoction group( 7. 8 g·kg~(-1)·d~(-1)). After continuous intragastric administration for 4 weeks,the body weight,body length and abdominal circumferences of all the rats were measured,the Lee' s obesity indexes was calculated,and the levels of fasting plasma glucose( FPG) and 2-hour plasma glucose( 2 hPG) in each group were measured. Serum insulin levels( FINS) and tumour necrosis factor-α( TNF-α) levels were detected by ELISA,and insulin sensitivity index( ISI) and insulin resistance index( IRI) values were calculated. The expressions of cysteinyl aspartate specific proteinase-1( caspase-1),gasdermin D( GSDMD),interleukin-1β( IL-1β) and interleukin-18( IL-18) m RNA and proteins in liver tissues were detected by Real-time PCR and Western blot. Immunofluorescence was used to detect the expressions of caspase-1 and GSDMD protein in liver tissues. Huanglian Wendan Decoction could effectively reduce the weight of model rats,improve abdominal obesity,FINS,IRI and ISI indexes and correct 2 hPG levels. Compared with blank group,TNF-α levels in serum and caspase-1,GSDMD,IL-1β and IL-18 expressions in liver tissue of model control group were increased significantly. Huanglian Wendan Decoction could effectively reduce TNF-α level in serum,regulate the expressions of caspase-1,GSDMD,IL-1β and IL-18 genes and proteins in liver tissues of IGT rats. The above results showed that the occurrence and development of " obesity-IR-abnormal glucose metabolism-type 2 diabetes mellitus" was closely related to inflammatory response and the classical pyroptosis pathway mediated by caspase-1/GSDMD. The mechanism of Huanglian Wendan Decoction in improving IR may be correlated with reduction of the level of inflammatory factors and the alleviation of the state of pyroptosis,and thus reverse the course of IGT.
Subject(s)
Animals , Rats , Diabetes Mellitus, Type 2 , Drugs, Chinese Herbal , Liver , PyroptosisABSTRACT
OBJECTIVE@#To observe the effect of electroacupuncture (EA) of " acupoint combination" on appetite, body fat, insulin sensitivity and central sensitivity of cholecystokinin (CCK) in obese rats with insulin resistance (IR), and to explore the mechanism of EA on improving obesity with insulin resistance.@*METHODS@#Among the fifty 8-week-old healthy SPF male Wistar rats, 10 rats were randomly selected and fed with normal diet; after 8 weeks, 8 rats were randomly selected as a normal group. The remaining 40 rats were fed with high-fat diet to establish the model of obsesity IR; after 8 weeks, 24 rats with successful model of obsesity IR were randomly divided into a model group, an EA group and a sham EA group, 8 rats in each group. Eight weeks after model establishment, the rats in the EA group were intervened with EA at "Fenglong" (ST 40), "Zhongwan" (CV 12), "Guanyuan" (CV 4) and "Zusanli" (ST 36), with continuous wave, in frequency of 2 Hz, and current intensity of 1 mA, for 10 min each time. The rats in the sham EA group were intervened with EA at the points 5 mm next to the acupoints used in the EA group and no electricity was given; the sham EA was given for 10 min each time. Both the treatments were given once every other day for 8 weeks. The Lee's index and food intake were observed before the intervention as well as 2 weeks, 4 weeks, 6 weeks and 8 weeks into intervention; after the intervention, serum insulin (INS) and glucose infusion rate (GIR) were detected; serum cholecystokinin (CCK) level was detected by ELISA; c-fos expression in the area postrema (AP) and nucleus tractus solitarius (NTS) of medulla oblongata was detected by immunohistochemistry.@*RESULTS@#Before the intervention as well as 2 weeks, 4 weeks, 6 weeks and 8 weeks into intervention, the Lee's index and food intake in the model group were higher than those in normal group (<0.01). The Lee's index (6 weeks and 8 weeks into intervention) and food intake (4 weeks, 6 weeks and 8 weeks into intervention) in the EA group were lower than those in the model group and the sham EA group (<0.05, <0.01). After intervention, compared with the normal group, serum level of INS was increased (<0.01), while GIR, serum CCK level, c-fos expression in AP and NTS in the model group were decreased (<0.01, <0.05). Compared with the model group and the sham EA group, serum level of INS in the EA group was decreased (<0.01), and GIR, serum CCK level, c-fos expression in AP and NTS were increased (<0.01, <0.05).@*CONCLUSION@#EA of " acupoint combination" could effectively reduce appetite, body fat and enhance insulin sensitivity in obese rats with IR. The mechanism may be related to the regulation of central sensitivity of CCK.
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OBJECTIVE@#To explore the therapeutic effect and partial mechanism of electroacupuncture (EA) for patients with insulin resistance (IR) polycystic ovary syndrome (PCOS).@*METHODS@#Seventy patients with IR-PCOS were randomly divided into an EA group (36 cases, 5 cases dropped off) and a medication group (34 cases, 4 cases dropped off). The patients in the medication group were treated with oral administration of metformin hydrochloride, 500 mg each time, twice a day. The patients in the EA group were treated with EA (continuous wave, 2 Hz of frequency) at Zusanli (ST 36), Zhongwan (CV 12), Qihai (CV 6), Yishu (EX-B 3), Shenshu (BL 23), Pishu (BL 20), Ciliao (BL 32) for 30 min, three times a week. One menstrual cycle or 4 weeks were taken as a course of treatment, and 3 continuous courses were given. The follow-up was 3 months. The lipid metabolism indexes of triacylglycerol (TG), total cholesterol (TC), high-density lipoprotein (HDL) and low-density lipoprotein (LDL), homeostasis model assessment-insulin resistance index (HOMA-IR) and testosterone (T) in serum were compared before and after treatment, and the clinical effects of the two groups were evaluated during the follow-up.@*RESULTS@#The total effective rate was 67.7% (21/31) in the EA group and 60.0% (18/30) in the medication group, with no significant difference between the two groups (>0.05). After treatment, the levels of serum T, HOMA-IR, LDL, TG and TC were decreased significantly in the two groups (<0.01, <0.05), and HDL was increased significantly (<0.01); the levels of TC in the EA group after treatment was lower than that in the medication group (<0.05).@*CONCLUSION@#EA may adjust some dyslipidemia in patients to correct IR and improve endocrine disorder of PCOS, which had superior/similar effects to metformin.
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Female , Humans , Acupuncture Points , Electroacupuncture , Insulin Resistance , Polycystic Ovary Syndrome , TherapeuticsABSTRACT
Objective::To define the clinical efficacy of modified Taohe Chengqitang combined with colon hydrotherapy in patients with severe nonalcoholic fatty liver disease (NAFLD) accompanied by phlegm-heat stagnation syndrome and its mechanism. Method::Totally 100 patients with severe NAFLD by phlegm-heat stagnation syndrome were enrolled in the study.They were all given Shanzha Xiaozhi capsule.According to the random number table, the patients were randomly divided into the observation group (50 patients, colon hydrotherapy combined with traditional Chinese medicine) and the control group (50 patients, Shanzha Xiaozhi capsule alone). The observation period was 4 weeks.The therapeutic effect of colon hydrotherapy was verified through determinations of the liver function, blood lipid, insulin resistance index (IRI), controlled attenuation parameter (CAP), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) before and after treatment.The mechanism of colon hydrotherapy combined with modified Taohe Chengqitang was preliminarily analyzed based on changes of IR, TNF-α and IL-6. Result::Alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptadase (γ-GT), total cholesterol (TCH), triglyceride (TG), fasting plasma glucose (FPG), fasting insulins (FINS), IRI, CAP, TNF-αand IL-6 of NAFLD patients in both of two groups were significantly lower than those before treatment (P<0.01). ALT, AST, γ-GT, TCH, TG, FPG, IRI, CAP, TNF-α and IL-6 in observation group were significantly lower than those in the control group after treatment (P<0.01). FINS in observation group was significantly lower than that in the control group (P<0.05). Conclusion::Colon hydrotherapy combined with modified Taohe Chengqitang is an effective method for treating NAFLD accompanied by phlegm-heat stagnation syndrome.Its mechanism may be mainly correlated with the reduction of IRI, serum TNF-α and IL-6.The course of colon hydrotherapy, the therapeutic mechanism and the long-term efficacy need to be further studied in the future.
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Objective • To investigate the effects of ovarian granulosa cell androgen receptor splice variant insertion isoform (AR-ins) on the levels of sex hormones and insulin in patients with polycystic ovary syndrome (PCOS). Methods • Seventy-nine control patients (control group) of normal ovulation and one hundred and fifty-four PCOS patients (PCOS group) who received in vitro fertilization embryo transfer in the Assisted Reproductive Department of the International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine from Jun. 2017 to Jun. 2018 were recruited. The sociodemographic information, serum basic hormones and insulin data were collected according to medical history record, and the hormone levels were measured by chemiluminescence. The ovarian granulosa cells and follicular fluids of all the patients were collected. Nested PCR was used to identify the androgen receptor splice variant (AR SV) in the ovarian granulosa cells from PCOS patients, and then they were divided into two groups, i.e., PCOS Wt group (n=39) and PCOS AR-ins group (n=115). The levels of sex hormones and insulin in follicular fluids of the three groups were detected by chemiluminescence. The free androgen index (FAI) was calculated. The sociodemographic information, the levels of sex hormones and insulin in serum and follicular fluid of the three groups were compared by One-way ANOVA, Welch's ANOVA, Kruskal-Wallis H test and chi-square test. Results • Compared with the control group, the levels of serum sex hormone-binding globulin (SHBG) were significantly lower in the PCOS AR-ins group (P=0.000), and the levels of serum total testosterone (TT) (P=0.000), insulin (P=0.001) and FAI (P=0.000) were significantly higher in the PCOS AR-ins group. Compared with the control group and the PCOS Wt group, the differences of SHBG, TT, insulin levels and FAI in the follicular fluid in the PCOS AR-ins group were statistically significant (all P<0.05). Conclusion • PCOS patients with AR-ins have high androgen and insulin levels in serum, and this effect is more pronounced in the ovarian local microenvironment.
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To investigate the hypoglycemic effects of baicalin, berberine, puerarin and liquiritin on the insulin resistance (IR) cells. The IR model of HepG2 cells was established by treatment with insulin and dexamethasone for 48 h. Glucose uptake, glycogen content and cell viability were detected with different concentrations of baicalin, berberine, puerarin, liquiritin in IR-HepG2 cells. Compared with IR model group, all of intervened groups significantly increased the glucose consumption, except for liquiritin groups and 1 μmol·L⁻¹ baicalin group. Moreover, 10, 20, 50 μmol·L⁻¹ baicalin, 5, 10, 20, 50 μmol·L⁻¹ berberine and 40, 80, 160 μmol·L⁻¹ puerarin significantly elevated glycogen content in IR-HepG2 cells. Liquiritin did not show obvious hypoglycemic effect. Compared with normal group, the mRNA expression levels of GLUT1 and GLUT4 were decreased in IR-HepG2 cells according to qPCR results. 5, 20 μmol·L⁻¹ berberine decreased the mRNA expression level of GLUT1 in IR-HepG2 cells, whereas 20, 40, 80 μmol·L⁻¹ puerarin significantly elevated the mRNA expression level of GLUT1. Moreover, 10, 20, 50 μmol·L⁻¹ baicalin and 20 μmol·L⁻¹ berberine increased the mRNA expression level of GLUT4. Whereas, 40, 80 μmol·L⁻¹ puerarin decreased the mRNA expression level of GLUT4. Western blot results suggested that 10, 20, 50 μmol·L⁻¹ baicalin significantly increased the protein expressions of GLUT2 and GLUT4, whereas 20, 40, 80 μmol·L⁻¹ puerarin significantly up-regulated GLUT1 and GLUT2 proteins. In addition, 20 μmol·L⁻¹ berberine increased the protein expressions of GLUT2 and GLUT4, whereas 10 μmol·L⁻¹ berberine up-regulated GLUT4 expression. The results preliminarily suggested that baicalin, berberine and puerarin have differentiated hypoglycemic effects, which accelerate glucose transport, increase glycogen synthesis, regulate glucose metabolism and improve hepatic IR.
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Humans , Berberine , Pharmacology , Flavonoids , Pharmacology , Glucose , Hep G2 Cells , Hypoglycemic Agents , Pharmacology , Insulin , Insulin Resistance , Isoflavones , PharmacologyABSTRACT
This paper aimed to investigate the hypoglycemic effect and relative mechanism of jatrorrhizine in insulin-resistance (IR)-3T3-L1 adipocytes. The 3T3-L1 preadipocytes were used to induce mature adipocytes, then the stable IR model was established with 1 μmol·L⁻¹ dexamethasone. The adipocytes were divided into normal group, IR model group, rosiglitazone positive group and jatrorrhizine group (0.5, 1, 5, 10, 20 μmol·L⁻¹). After different time points (12, 24, 30, 36, 48 h) treatment, glucose content of 3T3-L1 adipocytes was detected by the glucose oxidase peroxidase method and TG content was measured by glycerol phosphate oxidase method, whereas cell viability was detected by CCK-8 assay. Furthermore, the protein expression levels of insulin receptor substrate 2 (IRS2), phosphinositide-3-kinase regulatory subunit 1(PI3KR1), phosphorylated protein B [p-AKT (Ser473)], phosph-AMP-activated protein [p-AMPK (Thr172)], and glucose transporter type 4/1/2 (GLUT4/1/2) were detected by Western blot assay. The results showed that as compared with the normal group, the glucose consumptionwas significantly decreased in IR model group(<0.01); whereas 0.5, 1, 5, 10, 20 μmol·L⁻¹ jatrorrhizine and rosiglitazone group elevated IR-3T3-L1 cells glucose consumption (<0.01) at 36 h and 48 h administration as compared with IR group. The optimal administration time was 48 h for jatrorrhizine. 1, 5, 10, 20 μmol·L⁻¹ of jatrorrhizine decreased the TG content in 3T3-L1 adipocytes for 48 h administration (<0.05). The protein expression levels of IRS2, PI3KR1, p-AKT (Ser473), p-AMPK (Thr172), GLUT4/1/2 were significantly up-regulated by different concentrations of jatrorrhizine and rosiglitazone (<0.01). The results showed that jatrorrhizine increased glucose uptake with elevated glucose consumption, whereas reduced intracellular TG content in IR-3T3-L1 adipocytes. Moreover, it intervened classic insulin signal pathway IRS2/PI3KR1/p-AKT/GLUT4 and increase AMPK protein phosphorylation level for the activation of GLUT1/4 for insulin sensibility. Thus, jatrorrhizine could effectively regulate the GLUTs with multiple manners for hypoglycemic effect.
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Pigment epithelium-derived factor (PEDF ) was originally found fromretinal pigment epithelium. It is confirmed that it plays an anti-angiogenic and apoptosis role in diabetic microangiopathy. Recently ,PEDF has been found to have a close relationship with insulin resistance ,type 2 diabetes and diabetic macroangiopathy. But the mechanism is unclear. More and more researches focused on its role in oxidative stress. PEDF may become a new target for diabetes and diabetic complication treatment and also a predictive factor for the disease.
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Objective To investigate the relationships between plasma secreted frizzled-related protein (SFRP) 5 level with type 2 diabetes (T2DM ) and carotid atherosclerosis (CAS). Methods According to the results of carotid color Doppler ultrasound ,56 T2DM patients were divided into 28 with carotid atherosclerosis (CAS group) and 28 without carotid atherosclerosis (NAS group). 28 healthy volunteers served as normal control (NC group). Serum SFRP5 was measured by ELISA. Results The level of serum SFRP5 in T2DM patients was lower than NC group (114.36 ± 25.48) vs (48.19 ± 11.82) , (43.88 ± 8.19)pg/ml ,(P 0.05 ]. Pearson correlation analysis showed that serum SFRP5 was negatively correlated with age ,TG ,hsC-RP ,FPG ,FIns , HOMA-IR ,HbA1c ,TC ,LDL-C and BMI (P0.05). Multiple linear regression results showed that age ,FIns and HbA1 c were independent influencing factors of serum SFRP5. Conclusion SFRP5 may be a protective factor for T2DM by ameliorating insulin resistance which may provide a new clue for the prevention and treatment of T 2DM.
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Objective To explore the relation between serum nesfatin-1 ,tumor necrosis factor (TNF)-αand insulin resistance. Methods A total of 105 subjects were enrolled in this study and divided into two groups:patients with newly diagnosed type 2 diabetes mellitus (T2DM group,n= 64)and normal controls (NC group,n= 41). The fasting serum nesfatin-1 and TNF-αlevels were measured by enzyme-linked immuno sorbent assay (ELISA). FPG,HbA1 c,TG,TC,and FIns were also tested. BMI, HOMA-IR,HOMA-β,and ISI were calculated. Results Serum nesfatin-1 and TNF-αlevels were significantly higher in T2DM group than in NC group. Multiple linear regression analysis showed that the most significant influencing factors for nesfatin-1 were TNF-αand ISI(β= 0. 005、-6. 847,P<0. 05). The most significant influencing factors for TNF-αwas HbA1 c (β= 26. 652,P<0. 01). Conclusion Serum nesfatin-1 and TNF-αare significantly elevated in patients with newly diagnosed T2DM,which may influence the glucolipid metabolism through the signal pathway of insulin and play a role in the pathogenesis of T2DM and IR.
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Objective To investigate the effects of insulin resistance (IR)on hypoxia/reoxygenation (H/R)inj ury in PC1 2 cells and the protective effects of N-actyl-L-cystine (NAC)against H/R inj ury. Methods PC12 cells were treated with 100 nmol/L insulin to induce IR. The H/R injury model was established by Na2 S2 O4. CCK-8 assay was used to detect the cell viability. Glucose consumption was detected by glucose oxidase method. Activity of superoxide dismutase (SOD)was detected by xanthine oxidase method. The levels of malonaldehyde (MDA)were measured by thiobarbituric acid method. Flow cytometry was used to determine the apoptosis and mitochondrial membrane potential (MMP). Results High-insulin inhibited insulin-induced glucose uptake in PC12 cells without affecting the cell viability (P<0. 05). PC12 cells with IR exhibited lower SOD activity and higher levels of MDA (P<0. 05 ),and enhanced apoptosis and depolarization of MMP induced by H/R(P<0. 05). NAC neutralized these effects induced by IR(P<0. 05). Conclusion IR aggravates H/R injury in PC12 cells by enhancing oxidative stress and NAC reduces the H/R injury in PC12 cells with IR via inhibiting oxidative stress and stabilizing MMP.
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To observe the anti-hyperglycemic effect of Puerariae Lobatae Radix in hepatocyte insulin resistance(IR) models, and investigate its preliminary molecular mechanism. IR-HepG2 cell model was stably established with 1×10-9 mol•L⁻¹ insulin plus 3.75×10-6 mol•L-1 dexamethasone treatment for 48 h according to optimized protocol in our research group. After IR-HepG2 cells were treated with different concentrations(5%,10% and 15%) of Puerariae Lobatae Radix-containing serum, cell viability was detected by CCK-8 assay; the glucose consumptions in IR-HepG2 cells were separately detected at different time points (12, 15, 18, 21, 24, 30, 36 h) by using glucose oxidase method; intracellular glycogen content was detected by anthrone method; and the protein expression levels of leptin receptor (Ob-R), insulin receptor substrate-2 (IRS2), glucose transporter 1(GLUT1) and GLUT2 were detected by Western blot assay. The results showed that Puerariae Lobatae Radix-containing serum (5%, 10% and 15%) had no significant effect on IR-HepG2 cell viability; 5% and 10% Puerariae Lobatae Radix-containing serum significantly increased glucose consumption of IR-HepG2 cells (P<0.01) at 18, 21 and 24 h; 15% Puerariae Lobatae Radix-containing serum elevated the glucose consumption of IR-HepG2 cells at 15 h (P<0.05), and significantly elevated the glucose consumption at 18, 21, 24 and 30 h (P<0.01) in a dose-dependent manner. The optimized time of anti-hyperglycemic effect was defined as 24 h, and further study showed that Puerariae Lobatae Radix-containing serum could increase intracellular glycogen content after 24 h treatment (P<0.01), and up-regulate IRS2, Ob-R, GLUT1 and GLUT2 protein expression levels. Our results indicated that Puerariae Lobatae Radix-containing serum could achieve the anti-hyperglycemic effect through important PI3K/PDK signaling pathway partially by up-regulating the expression levels of Ob-R and IRS2, GLUT1 and GLUT2 in IR-HepG2 cells, accelerating the glucose transport into hepatocytes and increasing hepatic glycogen synthesis to enhance the anti-hyperglycemic effect of IR-HepG2 cells.
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This study aimed to explore the mechanism of Chinese traditional medicine, Kudzu root(Chinese name：Ge-Gen; Latin name： Puerariae Lobatae Radix) how to improving insulin resistance (IR) through the regulation of the glucose and lipid metabolism in the IR-3T3-L1 adipocytes. After the 3T3-L1 mouse preadipocytes were differentiated into mature adipocytes, IR model(IR-3T3-L1) was built with 1 μmol•L-1 dexamethasone treatment for 96 h. IR adipocytes were treated with different concentrations (5%,10% and 15%) of Ge-Gen containing serum (GG-CS)for 12 h or 24 h, whereas rosiglitazone group as positive control in this study. The glucose contents in cell culture supernatants were detected by glucose oxidase assay and the intracellular triglyceride (TG) contents were measured by glycerol phosphate oxidase assay respectively.The mRNA expression levels of PPARγ, ADPN, GLUT4, LPL, FABP4 and FASn gene were determined by real-time quantitative PCR(qPCR).Results showed that IR-3T3-L1 adipocytes significantly increased glucose consumption (P<0.01)and decreased TG contents (P<0.01) as compared with the normal control group, the glucose consumption significantly increased with the treatment of GG-CS (P<0.01) by dose-dependent and time-dependent manners,whereas the intracellular TG content was sigificantly decreased (P<0.01) by dose-dependent manner.qPCR analysis revealed that 10% and 15% GG-CS significantly up-regulated the mRNA expression level of PPARγ, ADPN and GLUT4 (P<0.01) with the same dose-dependent manner,whereas the GLUT4 mRNA expression was showed similar expression pattern with the treatment of 10% and 15% GG-CS (P<0.01).We also detected the mRNA expression levels of several important lipid-metabolizing enzymes such as LPL, FASn and FABP4 by PPARγ regulation. 15% GG-CS elevated LPL mRNA expression (P<0.05);10% and 15% GG-CS enhanced the FASn mRNA expression (P<0.01), whereas 5%,10% and 15% GG-CS down-regulated FABP4 mRNA expression (P<0.01). Together, our results indicated that GG could regulate the glucose and lipid metabolism to ameliorate IR with multi-target manners in 3T3-L1 adipocytes.
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Objective To investigate correlation of abdominal adipose tissue distribution and insulin resistance in T2DM. Methods A total of 128 T2DM patients were divided into two groups :obese (OG) group (n=66) and non‐obese (NOG) group (n=62). Spiral CT was used for the measurement of adipose tissue of the total area (TA) and visceral fat area (VA) at abdominal umbilical level lumbar vertebrae 4 ,5 plane in T2DM patients. Subcutaneous fat area (SA ) was calculated. General and biochemical characteristics were measured in both groups. Results WC [male(73.52 ± 0.88) vs (70.66 ± 0.92)cm ;female(83.22 ± 0.96) vs (78.98 ± 0.98)cm] ,BMI [male(28.85 ± 3.45) vs (25.11 ± 4.36)kg/m2 ;female (28.23 ± 3.48) vs (25.05 ± 3.89)kg/m2 ] ,SBP [male(158.23 ± 8.25) vs (112.25 ± 7.25)mmHg ;female (154.25 ± 6.32) vs (109.68 ± 8.02)mmHg] ,DBP [male(95.36 ± 5.26) vs (80.69 ± 7.25)mmHg ;female (92.45 ± 4.36) vs (80.26 ± 6.48)mmHg] ,FPG [male(9.85 ± 2.89) vs (7.03 ± 2.88)mmol/L ;female (9.75 ± 2.65) vs (7.39 ± 2.98)mmol/L] ,FIns [male(11.25 ± 3.45) vs (7.02 ± 2.43)mIU/L ;female (11.02 ± 3.58) vs (7.18 ± 2.69)mIU/L] ,HbA1c [male(7.36 ± 1.36)% vs (5.21 ± 0.37)% ;female(7.68 ± 1.22)% vs (5.32 ± 0.42)% ] ,TG [male(5.98 ± 1.52) vs (3.02 ± 0.89)mmol/L ;female(5.78 ± 1.26) vs (2.98 ± 0.92)mmol/L] ,TC [male(8.02 ± 1.28) vs (4.39 ± 0.98)mmol/L ;female(7.98 ± 1.13) vs (4.23 ± 0.89)mmol/L] ,LDL‐C [male(9.12 ± 0.58) vs (4.21 ± 0.86)mmol/L ;female(8.96 ± 0.78) vs (4.18 ± 0.92)mmol/L] ,SUA [male(83.63 ± 21.64) vs (72.98 ± 12.25)μmol/L ;female(83.98 ± 19.78) vs (71.98 ± 11.98)μmol/L] ,C‐RP [male(5.96 ± 1.25) vs (2.32 ± 0.42)mg/L ;female(5.05 ± 1.32) vs (2.52 ± 0.56)mg/L] ,HOMA‐IR [male(4.25 ± 1.12) vs (2.25 ± 1.12);female(4.36 ± 1.42) vs (2.12 ± 1.02)] ,TA [male(50.68 ± 9.12) vs (30.96 ± 3.26)cm2 ;female(47.23 ± 4.23) vs (26.98 ± 3.02)cm2 ] , VA [male(19.78 ± 5.42) vs (10.59 ± 4.69)cm2 ;female(17.02 ± 3.96) vs (8.45 ± 3.78)cm2 ] ,SA [male (30.91 ± 6.02) vs (18.96 ± 5.78)mm2 ;female(28.25 ± 4.23) vs (17.25 ± 4.62)mm2 ]and VA/SA [male (0.72 ± 0.22)% vs (0.42 ± 0.18)% ;female(0.58 ± 0.17)% vs (0.32 ± 0.12)% ] were significantly higher in OG group than in NOG group. T2DM course [male(2.36 ± 0.58) vs (2.62 ± 0.78)years ;female (2.38 ± 0.62) vs (2.82 ± 0.82)years] ,HDL‐C [male(0.98 ± 0.21) vs (2.28 ± 0.78)mmol/L ;female(0.96 ± 0.32) vs (2.19 ± 0.82)mmol/L] and HOMA‐β[male(28.22 ± 9.34) vs (82.22 ± 31.25);female(28.02 ±8.02) vs (81.36 ± 28.36)] were lower in OG group than in NOG group(P< 0.05). Spearson correlation analysis showed that HOMA‐IR was positively associated with TG ,SUA ,TA ,VA ,SA ,VA/TA ,SA/TA and VA/SA. Logistic multiple regression analysis showed that TG ,SA ,TA and VA/TA were risk factors for insulin resistance in T2DM patients. Conclusion Abdominal fat distribution is closely related to IR in T2DM patients.
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[Summary] Considerable efforts have been made to understand the cellular and molecular mechanisms of metformin ,a potent antihyperglycemic agent now recommended as the first‐line treatment of patients with T2DM. The main effect of this drug is to reducethe hepatic glucose output ,primarily through inhibition of the mitochondrial respiratory chain complex Ⅰ and then activating AM P‐activated protein kinase (AMPK) ,which provide a generally acceptable mechanism for the action of metformin on hepatic gluconeogenesis. Beyond its effect on glucose metabolism ,metformin has been reported to improve ovarian function of patients with polycystic ovary syndrome (PCOS) and to reduce the risk of microvascular and macrovascular complications in patients with T2DM. In addition ,metfomin has also recently been suggested as an adjuvant treatment of cancer. Here we reviewed the progress of mechanism research and clinical application of metformin.
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Aim: Obesity and family history of Type 2 Diabetes mellitus are the major risk factors for the development of type 2 Diabetes mellitus in youth. The purpose of this study is to investigate the association between serum ferritin and insulin resistance in healthy young obese with and without family history of type 2 Diabetes mellitus. Place and Duration of Study: Department of Biochemistry, Rajarajeswari Medical College and Hospital, Kambipura, Bangalore, Karnataka, India, for eight months period in the year 2012. Material and Methods: A small group study was undertaken in 90 students who were in the age group of 17-22 years. The study population was divided into two groups based on body mass index, Group I /non-obese group (n=46) and Group II/ overweight & obese group (n=44). Fasting and postprandial blood glucose, Serum ferritin, serum insulin and lipid parameters were estimated and Homeostasis Model Assessment-Insulin resistance (HOMA-IR) was calculated for all the ninety students. Results: Statistically significant differences in Total cholesterol (p=0.05), Triglycerides (p=0.05), serum insulin (p<0.01) and HOMA (p<0.01) were observed between the two groups. Mean serum ferritin values were increased in group II (overweight/obese) but not statistically significant. Serum insulin and serum ferritin showed a significant correlation with BMI for the whole study group. Serum ferritin and insulin levels significantly correlated with waist to hip ratio in students with family history of Type 2 Diabetes mellitus as against individuals without family history. 59% of the obese students with family history of Diabetes mellitus had insulin resistance. Conclusion: Our study has shown that a significant proportion of obese students with family history of type 2 diabetes mellitus had insulin resistance and elevated levels of ferritin, highlighting the importance of early screening for obesity associated co morbidities like metabolic syndrome, Type 2 Diabetes mellitus and cardiovascular diseases in these individuals