| Objective1 To observe the therapeutic effect of Bushen tiaogan Jiangzhuo Recipe on obese patients with polycystic ovary syndrome(PCOS).The clinical efficacy of Bushen tiaogan Jiangzhuo recipe was preliminarily evaluated by its effects on ovulation,sex hormone level and glucose and lipid metabolism.2 Based on AMPK/PGC-1a signal pathway,observe the effects of Bushen tiaogan Jiangzhuo Recipe on autophagy and lipid metabolism of ovarian granulosa cells in obese PCOS model rats,and further explore the therapeutic target and mechanism of Bushen tiaogan Jiangzhuo Recipe on obese PCOS.Method1 Clinical studyUsing the self-control research method before and after treatment,83 obese PCOS patients with phlegm and blood stasis syndrome were collected.They were given Bushen tiaogan Jiangzhuo Recipe for three consecutive menstrual cycles(menstrual period without stopping).The general conditions,basal body temperature,sex hormone level,glucose and lipid metabolism indexes and symptom scores before and after treatment were compared,and the pregnancy rate of patients with pregnancy needs was followed up.2 Experimental researchThe obese PCOS rat model was established by intragastric administration of letrozole and high-fat diet.The rats were divided into blank group,model group,low,medium and high dose groups of Bushen tiaogan Jiangzhuo formula and metformin group.After drug intervention,the ovarian morphology of rats in each group was observed by HE staining;Serum neutral hormone,insulin,leptin and adiponectin were detected by ELISA;The expression levels of AMPK,PGC-1a and so on in ovary,white fat and brown fat were detected by real time PCR and Western blot,and the differences of expression levels were analyzed.Result1 Clinical observation of Bushen tiaogan Jiangzhuo Recipe on obese patients with PCOS(1)Patients with biphasic pregnancy or ovulation accounted for 18.1%;Patients with biphasic BBT accounted for 53%,patients with atypical biphasic accounted for 8.4%,and patients with monophasic basal body temperature accounted for 20.5%.The total effective rate was 79.5%.(2)Body mass index and waist hip ratio:after treatment,there was significant difference between the body mass index and waist hip ratio before treatment(P<0.01).(3)Sex hormone level:after treatment,the testosterone level of patients was significantly different from that before treatment(P<0.01);LH/FSH showed no statistical significance before and after treatment(P>0.05).(4)Blood lipid level:after treatment,there were significant differences in low-density cholesterol,high-density cholesterol,total cholesterol and triglyceride(P<0.01).(5)Insulin homeostasis model:after treatment,the insulin homeostasis model of patients was significantly different from that before treatment(P<0.01).(6)Symptom score:after treatment,the symptom score of patients was significantly higher than that before treatment(P<0.01).(7)Safety observation:no adverse events occurred before,after and during treatment.2 Effect of Bushen tiaogan Jiangzhuo Recipe on AMPK/PGC-1a pathway in obese PCOS rat model(1)Effects of on ovarian morphology,insulin and sex hormone levels① Compared with the blank group,the number of follicles in the model group increased significantly,but most of them were cystic vacuoles without oocyte development;Compared with the model group,the medium dose group had mature oocyte development.② BMI:compared with the blank group,there was significant difference in the model group(P<0.01);Compared with the model group,the results of the medium dose group,high dose group and metformin group showed significant difference(P<0.01),and the low dose group showed significant difference(P<0.05).③ Free testosterone:compared with the blank group,there was significant difference in the model group(P<0.01);Compared with the model group,the low,medium and high dose groups of traditional Chinese medicine showed significant difference(P<0.01),and the metformin group showed significant difference(P<0.05).④ LH/FSH:compared with the blank group,the model group had no statistical significance(P>0.05);Compared with the model group,there was no significant difference between the low,medium and high dose groups and metformin group(P>0.05).⑤ Fins:compared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,there was no significant difference between the low-dose group and the medium dose group(P>0.05),and there was significant difference between the high-dose group and metformin group(P<0.01).(2)Expression of AMPK/PGC-1a in model rat ovary① Gene expression.AMPK mRNA:compared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,the middle dose group and metformin group showed significant statistical significance(P<0.01),while the low dose group and high dose group had no statistical significance(P>0.05).PGC-la mRNA:there was significant difference between the model group and the blank group(P<0.05);Compared with the model group,the middle dose group and metformin group had significant statistical significance(P<0.05),while the low dose group and high dose group had no statistical significance(P>0.05).② Protein expression.AMPK protein:there was significant difference between the model group and the blank group(P<0.05);Compared with the model group,there was significant difference between the low-dose group and the medium dose group(P<0.05),and there was no significant difference between the high-dose group and metformin group(P>0.05).PGC-1a protein:compared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,the low-dose group showed no statistical significance(P>0.05),the medium dose group and high-dose group showed significant statistical significance(P<0.01),and the metformin group showed significant difference(P<0.05).(3)Study on lipid metabolism in model rats① Leptin:compared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,there was significant difference in the low,medium and high dose groups of traditional Chinese medicine(P<0.01),and there was no significant difference in the metformin group(P>0.05).② Adiponectin:compared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,there was no statistical significance in the low-dose group(P>0.05),but there was significant difference in the medium dose group,high-dose group and metformin group(P<0.05).③White fat Browning:In terms of gene level,AMPK-compared with the blank group,there was significant difference in AMPK in white fat in the model group(P<0.01);Compared with the model group,the middle dose group of traditional Chinese medicine showed significant statistical significance(P<0.01),the low and high dose groups of traditional Chinese medicine showed significant statistical significance(P<0.05),and the metformin group showed significant difference(P<0.05).PGC-1a-compared with the blank group,there was significant difference in the model group(P<0.05);Compared with the model group,there was significant difference between the medium dose group and metformin group(P<0.01),and there was significant difference between the low dose group and the high dose group(P<0.05).UCP-1-compared with the blank group,there was significant difference in the model group(P<0.05);Compared with the model group,the low-dose group and the medium dose group showed significant statistical significance(P<0.01),but there was no significant difference between the high-dose group and metformin group(P>0.05).In terms of protein level,there was significant difference between AMPK-and blank group(P<0.01);Compared with the model group,the medium dose group of traditional Chinese medicine showed significant statistical difference(P<0.01),the metformin group showed significant difference(P<0.05),and the other groups had no statistical significance(P>0.05).PGC-1a-compared with the blank group,the model group had significant statistical difference(P<0.01).Compared with the model group,the low and medium dose groups of traditional Chinese medicine showed significant statistical difference(P<0.01),and the high dose group of traditional Chinese medicine and metformin group showed significant difference(P<0.05).UCP-1-compared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,the middle dose group of traditional Chinese medicine showed significant difference(P<0.01),the high dose group of traditional Chinese medicine showed significant difference(P<0.05),and the low dose group of traditional Chinese medicine and metformin group showed no significant difference(P>0.05).④ Brown fat activation:In terms of gene level,AMPK-compared with the blank group,there was significant difference in the model group(P<0.05);Compared with the model group,the middle dose group had significant statistical difference(P<0.01),while the values of low dose group,high dose group and metformin group had no statistical significance(P>0.05).PGC-1acompared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,the difference was statistically significant in the medium dose group(P<0.05),but there was no significant difference in the low dose group,high dose group and metformin group(P>0.05).UCP-1-compared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,the middle dose group of traditional Chinese medicine had statistical significance(P<0.05),and the other groups had no statistical significance(P>0.05).In terms of protein level,AMPK-compared with the blank group,there was significant difference in the model group(P<0.05);Compared with the model group,there was significant difference between the low-dose group of traditional Chinese medicine(P<0.05),the middle dose group of traditional Chinese medicine and metformin group(P<0.01),and there was no difference between the high-dose group of traditional Chinese medicine(P>0.05).PGC-1a-compared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,there was significant difference in the low,medium dose group and metformin group(P<0.01),but there was no significant difference in the high dose group(P>0.05).UCP-1-compared with the blank group,the model group had significant statistical difference(P<0.01);Compared with the model group,the expression in the middle dose group of traditional Chinese medicine was statistically significant(P<0.01),but there was no statistical significance in the low and high dose groups of traditional Chinese medicine and metformin group(P>0.05).Conclusion1 The results of clinical research show that Bushen tiaogan Jiangzhuo decoction can significantly improve ovulation,weight loss,waist hip ratio,androgen level and glucose and lipid metabolism in obese PCOS patients.2 The experimental results show that Bushen tiaogan Jiangzhuo recipe can reduce the body weight,free testosterone,insulin and leptin of obese PCOS model rats,increase adiponectin,induce white fat browning and activate brown fat by activating AMPK/PGC-1a pathway,which may improve ovulation by promoting the autophagy of ovarian granulosa cells. |
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