Effect Of Jinlida On Ovarian Function In Rat Model Of Polycystic Ovary Syndrome And Its Mechanism

OBJECTIVE:This study was designed to investigate the mechanism of Jinlida granules in improving insuLin resistance in rats with polycystic ovary syndrome through PI3K/AKT/GLUT4 pathway,in order to guide the clinical treatment of PCOS patients.METHODS:Fifty-one-day Spragu Dawley(SD)female rats with 7 days of adaptive feeding were randomly divided into blank group and model group,including 10 blank groups and 40 model groups.The basic weight of the mouse was weighed before the beginning of the experiment.The model group was given subcutaneous injection of dehydroepiandrosterone(DHEA)6mg/100g/day +0.2mL soybean oil for 20 days to establish the PCOS rat model.The blank group was given subcutaneous injection of the same dose of oil.After the modeling was completed,the vaginal smears of the two groups were observed to judge the estrous cycle of the rats.Eight rats were randomy selected from the model group,and the ovarian tissue was taken for pathological section to judge the resuLts of PCOS modeling.Peritoneal glucose tolerance test(IPGTT)was performed in two groups of rats.Fasting blood glucose(FBG)and fasting insuLin(FINS)were measured.HOMA-IR index was calcuLated to evaluate insuLin resistance.After the modeling,the model group was randomly divided into the model group,metformin group,zinlida group and zinlida+metformin group,with 8 mice in each group.Gastric gavage was given to metformin 0.265g/(kg.d),zinlida 50mg/(kg.d),metformin 0.265g/(kg.d)+ Jinlida 50mg/(kg.d),respectively,while the blank group was given the same amount of normal saline.Each group was administered once a day for 28 days and the body weight was recorded once a week.After the end of the experiment,the IPGTT test was performed every other day to detect blood glucose and FINS at 0 min,60 min,and 120 min,and the HOMA-IR index was calcuLated.Serum testosterone(T),follicle-stimuLating hormone(FSH),luteinizing hormone(LH),anti-miller tube hormone(AMH),triglyceride(TG),total cholesterol(TC),low-density lipoprotein(LDL-C)and high-density lipoprotein(HDL-C)were detected by ELISA.Observation of pathological changes of ovary in rats of each group by HE staining.The expressions of PI3K mRNA,AKT mRNA and GLUT4 mRNA in liver and ovary of each group were detected by semi-quantitative reverse transcriptase chain reaction(qRT-PCR),while the levels of p-PI3K,p-AKT and GLUT4 in liver and ovary of each group were detected by Western-Blot method.RESULTS:At the end of modeling,the body weight,FSH,LH/FSH,E2 and AMH of the model group were higher than those of the blank group(P<0.05);serum LH and T were significantly higher than the normal group(P<0.01).The results of vaginal smear showed that the rats in the model group lost their estrous cycle changes.The results of HE staining in ovarian tissue indicated that the ovary of the model group showed polycystic changes,the follicular cavity increased,the tunica albuginea thickened,the atresia follicles increased,and the granule cell layer decreased.The gap between the follicular cells increases.The FBG,PBG 60',PBG 120',FINS and HOMA-IR of the model group were significantly higher than the blank group(P<0.01).After drug intervention,there was no significant difference in body weight between the groups(P>0.05).The level of PBG 60',FINS and HOMA-IR was significantly lower in the Jinlida group than in the model group(P<0.01);metformin Group and Jinlida + metformin group can significantly reduce PBG60',PBG120',FINS,HOMA-IR,(P<0.01).The expression levels of PI3K,AKT,GLUT4 mRNA in liver and ovary of the model group were significantly lower than those of the blank group(P<0.01);The expression of PI3K mRNA in liver and the expression of PI3K,AKT,and GLUT4 mRNA in the ovary can be increased in the Jinlida group(P<0.05).After combined with metformin,it can significantly increase the expression of PI3K and GLUT4 mRNA in the liver(P<0.01),increase the expression of AKT mRNA in the liver(P<0.05).It also can significantly increase the expression of PI3K,AKT and GLUT4 mRNA in ovary compared with the model group(P<0.01),and all of them are higher than that of the metformin group alone.The expression levels of p-PI3K,p-AKT and GLUT4 protein in liver and ovary of the model group were significantly lower than those of the blank group(P<0.01).Compared with the model group,the levels of p-PI3K and p-AKT in the liver,as well as p-PI3K and GLUT4 in the ovary were significantly increased in the Jinlida group(P<0.05),no statistical difference was found between liver GLUT4 and p-AKT in the ovary,but both showed an increasing trend.After combined with metformin,expression of p-PI3k,p-akt and GLUT4 proteins in liver and ovary increased significantly(P<0.01).The results of sex hormones showed that the levels of FSH,T,LH/FSH and AMH in the model group were higher than those in the blank group(P<0.05),and the levels of LH were significantly higher than those in the blank group(P<0.01).The levels of FSH,LH,T and AMH in Jinlida group were significantly lower than those in the model group(P<0.05).When combined with metformin,Jilida can significantly reduce the level of LH(P<0.01)and reduce AMH,compared with the model group(P<0.05).After the intervention of various drugs,ovarian HE staining showed that the polycystic changes of the ovary were significantly reduced under the microscope.In the metformin group and the metformin + Jilida group,the number of granulosa cell layers was reduced and expanded follicles with cystic shape were observed.In terms of blood lipids,TG level in the model group was higher than that in the blank group(P<0.05),and serum TG level could be reduced in each drug intervention group(P<0.05).Conclusions:Jinlida granules can improve polycystic changes and insulin resistance in PCOS rats,and it is more effective in improving insulin resistance when combined with metformin.Jinlida granules may improve the insulin resistance of PCOS rats through PI3K/AKT/GLUT4 pathway;Jinlida granules can reduce the levels of LH,T and AMH in model rats and improve the polycystic changes of ovary,while Jinlida granules can improve the hypertriglyceridemia status of PCOS rats.At the same time,Jinlida granules can reduce hypertriglyceridemia in PCOS rats.