| Objective In this study, the patients with polycystic ovary syndrome (PCOS) of kidney deficiency and phlegm stasis type and insulin resistance (IR) were treated with Shouwujiangqi decoction in the clinic. The clinical symptoms, reproductive hormones and glucose metabolism before and after treatment were objectively evaluated. In addition, the effects of Shouwujiangqi decoction on expressions of insulin receptor substrate-1(IRS-1) and phosphoinositide3-kinase (PI-3K) p85a subunit mRNAs and proteins, and phosphorylation of insulin receptor substrate-1serine307(IRS-1Ser307) in the livers and ovarian tissues of rats with PCOS and IR were determined. The efficacy and mechanisms of Shouwujiangqi decoction on patients with PCOS of kidney deficiency and phlegm stasis type and IR were explored. The experimental basis of Shouwujiangqi decoction in the treatment of PCOS with IR was provided.Method1. Clinical part:99patients meeting the standards formulated by this research institute were collected and randomly divided into the control group (western medicine group) and the observation group (western medicine plus Shouwujiangqi decoction). The control group included52patients and the observation group included47ones. Their heights, weights, serum FSH, LH, T, FBG, FINS, HOMA index, TG and Tc were measured. Their clinical symptoms such as menstruation, hirsutism and acne as well as the improved status of above symptoms within3months after treatment were recorded. The situation of ingravidation was observed for infertility patients.2. Experimental part:Prasterone sodium sulfate combined with HCG and high fat diet was used to establish rat PCOS-IR model. The rats were randomly divided into the model group, the treatment group, and the normal control group. The treatment group were treated with Shouwujiangqi decoction (high-dose, medium dose and low dose group) and metformin. The normal control group were fed with equal volume of double-distilled water. During the treatment, the treatment groups and model ones were continuously given with modeling drugs for stimulation and fed with high fat diets. The normal control group was fed with normal diets for21days.. On the18th day of the intervention, the tail vein bloods were extracted for intravenous glucose tolerance test (IGTT) and fasting blood glucose with the glucometer. Three days after IGTT, all rats were weighed. The bloods were taken. The serum FSH, LH and T were detected by radioimmunoassay. The fasting insulin was measured by ELISA. The rats were sacrificed. The liver, both ovaries and surrounding adipose tissues were removed. The gross anatomies of ovaries were observed. Both ovaries and their surrounding adipose tissues were weighed. The ovarian tissue morphology was observed under a light microscope. IRS-1, PI3Kp85a subunit mRNA and protein expressions in liver and ovarian tissues in each group were detected respectively by real-time quantitative PCR (Real-time PCR) and protein immunoblotting (Western blotting). The phosphorylated IRS-1Ser307proteins in liver and ovarian tissues in each group were detected by Western blotting.Results1.Shouwujiangqi decoction can significantly improve the patients’clinical symptoms such as irregular menstruation, soreness and weakness of waist and knees, dizziness and tinnitus, weakness and tiredness, thoracic or abdominal fullness, greasy mouth and excessive phlegm, breast distending pain, and distending pain in lower abdomen etc. The efficacy in improvement of TCM symptoms in the treatment group was significantly better than that in the control group (P<0.05). The efficacy in improvement of physical signs, including the reduction of BMI and the treatment of hirsutism, in both the treatment and control groups was similar (P>0.05). In the treatment of acnes, the efficacy in the treatment group was better than that in the control group (P <0.01). In terms of the pregnancy rate of infertility patients, the efficacy in the treatment group was better than that in the control group (P<0.05).The clinical total effective rate in treatment group was better than that in the control group (P<0.05).The serum LH, LH/FSH, T value, insulin and HOMA index in the treatment group were significantly reduced than those in the control group; There were no significant differences in fasting blood glucose, triglycerides and total cholesterol between both groups (P>0.05)2. The gross anatomies of ovaries:The colors and lusters of ovaries in the normal control group were ruddy. The ovarian surfaces in model control group were pale, and their volumes were increased. The ovarian weights in the model control group were significantly increased when compared with those in the normal control group (P<0.05).3. The ovaries under light microscopy:the follicle and multiple corpus luteum formations in different developmental stages were observed in the ovarian tissues in the normal control group. The granular cell layers in mature follicles were in alignment; the ovarian surface in the model control group showed multiple dilated cystic follicles. The oocytes or corona radiata disappeared within follicles. The granular cell layers were decreased, and there were fewer corpus luteum formations. After treatment, the dilated cystic follicles in ovarian tissues of rats were reduced. The granular cell layers were thickened, and corpus luteums were increased, especially in Shouwujiangqi decoction high dose group and metformin group.4. Serum T, fasting glucose, fasting insulin and HOMA index were significantly reduced in Shouwujiangqi decoction high-dose group (P<0.01).5. The levels of IRS-1, PI3Kp85a subunit mRNA and protein in liver and ovarian tissues were significantly lower in the model group than in the normal group (P<0.01), while their levels were significantly higher in the treatment group including Shouwujiangqi decoction high dose and metformin subgroups than in the model group (P<0.01).6. The levels of IRS-1Ser307proteins in liver and ovarian tissues of rats was significantly higher in the model group than in the normal group, while their levels were significantly lower in the treatment group including Shouwujiangqi decoction high dose and metformin subgroups than in the model group(P<0.01).Conclusion1.Shouwujiangqi decoction can significantly improve clinical symptoms of polycystic ovary syndrome patients with insulin resistance, and increase the reproductive hormones, glucose metabolism as well as the pregnancy rate.2. Prasterone sodium sulfate combined with HCG and high fat diet-induced pathological model of rat with PCOS and IR is successfully established. The researches have been performed in aspects of the ovarian morphology and endocrinology in the model. The results indicate that the pathological morphology, the reproductive hormones as well as the indicators of glucose metabolism are especially similar to those in PCOS patients with IR. Therefore, it is considered that the model is an ideal animal model to study the PCOS patients with IR.3. It is found in the experiment that Shouwujiangqi decoction high-dose group can improve the ovarian polycystic degenerations and ovulation disorders of polycystic ovary rats with insulin resistances, and decrease levels of reproductive hormones and glucose metabolism. Moreover, and Shouwujiangqi decoction can increase IRS-1and PI3K p85mRNA and protein expressions in liver and ovarian tissues of PCOS rats with IR, and inhibit IRS-1Ser307phosphorylation, thereby regulate the insulin signal transduction. These might be the mechanism of Shouwujiangqi decoction to improve the clinical syndromes of PCOS with IR patients. |
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