| Objective: Based on the establishment of PCOS(polycystic ovarian syndrome) rat model, this paper focuses on the effects of Kidney Tonifying and Phlegm Removing Prescription for the level of hormone in rats and its hypoglycemic effect, and evaluating the model animals’ reproductive phenotype and metabolic phenotype by measuring estrous cycle, OGTT tests etc. To explore the mechanism of action of TCM(traditional Chinese medicine) Formula of Kidney Tonifying and Phlegm Removing Prescription in treatment of PCOS, by comparing the changes of hormone levels and glucose metabolism between the intervention group and the control group.Method: 95 SD 21-day-old rats were randomly divided into pathological model group with 70 rats and the normal group with the other 25 rats. The 70 rats in the pathological model group were given subcutaneous injection of dehydroepiandrosterone(DHEA) 6mg / 100 g of body weight plus 0.2ml injection oil, and the other 25 rats in the control group were given subcutaneous injection of 0.2ml of injection oil for continuous 20 days. DHEA-induced rat model of PCOS were established. After modeling, each 10 rats were randomly selected from the normal group and the model group to make an assessment, the rest was used to do experimental drug intervention.The remaining 75 rats were randomly divided into normal control group, model control group, metformin group, TCM low-dose group and TCM high-dose group. The normal control group and the model control group received by gavage for an equal volume of 0.5% sodium carboxymethyl cellulose; the metformin group were given by gavage for 200 mg / kg metformin; the TCM low-dose group were given by gavage for 20 ml / kg(16.6g crude drug / kg);while the TCM high-dose group were given by gavage for 20 ml / kg(33.2g crude drug / kg).After each group of rats were fed continuously for 28 days, by assessing the OGTT and the estrous cycle, and monitoring FSH,LH, when the rat stays in the same estrous cycle, anesthesia with 10 percent of chloral hydrate, take 5ml blood from abdominal aorta, get the serum of centrifugation, keep portions in the refrigerator(-80℃)for reservation; Measuring the length, width, thickness of the rats’ ovary and weighed, placed in 4% paraformaldehyde solution reserved for test.Result: 1.General condition observation during the experiment: after continuous 20 days’ subcutaneous injection of DHEA, the model rats appeared reduced activity, increased diet and drinking water, and the increased urine volume etc., without obvious weight gain. While the weight of the rats in normal group increased gradually, mental symptoms and activated as usual. After gavage, the rats in metformin group, TCM low-dose group, TCM high-dose group appeared decreased body weight, diet and drinking water reduced than before, the state of mind was better than giving a gavage.2. Model evaluation: ①SD rats were found the irregular estrous cycle after a vaginal smear, and continuously stayed in estrogen interphase; the normal group set the day making a vaginal smear as estrus, and enters the post-estrus after 24 hours, vaginal smears performed regular estrous cycle.②Serum LH levels in the model rats was significantly higher than the normal group, P <0.01, it shows a significant difference; the normal group FSH compared with the model group, P> 0.05, without significant difference; the model rats LH / FSH compared with the normal group, P <0.01, which is significantly different.③Ovary volume of the model group was significantly higher than the normal group, P <0.01, with significant difference; Ovary weight of the model group compared with the normal group, P <0.05, which possess statistically significant.④After the analysis of variance on the weight of rats, P> 0.05, it showed that there was no significant difference between the control group and the model group.⑤GLU model group and HOMA-IR were significantly increased(P <0.01), AUC levels increased more than the normal group(P <0.05).3.Evaluation after treatment: ①By monitoring the vaginal smear, the rats in the normal group ovulated regularly; the model group appeared cycle disorder and ovulated irregularly; After intervention, the metformin group, TCM low-dose group and TCM high-dose group were ovulation obstacle ratio was 28.57%, 21.43% and 7.14%. TCM high-dose group were significantly higher ovulation obstacle cure rates than the metformin group and TCM low-dose group.②The serum LH level of the normal group, metformin group, TCM low-dose group and TCM high-dose group was significantly lower than the model group, P <0.01, was significantly different. TCM low-dose group’s LH was higher than metformin group(P <0.05), high dose group was significantly lower than metformin group(P <0.01). Normal group, metformin group, serum FSH level Chinese low-dose group had no significant change compared with the model group, with statistically significant(P <0.01) compared to the high dose group and model group. With a statistically signif icant normal group, metformin group, serum medicine low dose group and high dose group LH / FSH levels compared with the model group, P <0.01,. High dose group an d the metformin group, significantly lower than the model control group(P <0.01)③Normal rats ovarian volume compared with the model group(P <0.05), with a statistically significant; compared with the model group, the ovarian volume of the metformin group, TCM low-dose group and high-dose group with statistically significant(P<0.01); TCM low-dose group compared with the metformin group, ovarian volume decreased, with a statistically significant(P <0.05), while the high-dose group was significantly lower, with a statistically significant(P <0.01).Compared with the model group, the ovary weight of the normal group and the TCM low-dose group possessed statistically significant(P <0.05), between the metformin group and the model group P> 0.05, without statistically significant, TCM high-dose group with a statistically significant(P <0.01) compared with the model group. TCM low-dose group compared with the metformin group was not statistically significant(P> 0.05), TCM high-dose group was statistically significant(P <0.05).④After analysis of variance, the body weight of the normal group, metformin group, the TCM low-dose group and the model group was no obvious difference(P>0.05); between the TCM high-dose group and the model group turned out obvious difference(P<0.01), compared with the metformin group, P <0.05, with statistically significant.⑤Compared with the model group, the level of GLU, AUC and HOMA-IR of the normal group, metformin group, TCM low-dose group, TCM high-dose group were obviously lower, with a statistically significant(P <0.01). GLU of TCM low-dose group and TCM high-dose group compared with the model group, without statistically significant.AUC of TCM low-dose group, TCM high-dose group compared with the metformin group, P <0.05, showed statistically significant. HOMA-IR of TCM low-dose group, TCM high-dose group were with statistically significant compared with the metformin group(P <0.01).Conclusion: 1.This experiment successfully established the rat model of DHEA-induced PCOS, and make a study on POCS from endocrine and glucose metabolism, find that changes of its endocrine and glucose metabolism is very similar to the POCS, which exist hyperinsulinemia. Therefore, this model is more than a mature and ideal PCOS animal model.2. Kidney Tonifying and Phlegm Removing Prescription can reduce LH, LH / FS H levels and improve PCOS endocrine patient, thereby restoring ovarian function, pro mote normal ovulation.3. The formula of Kidney Tonifying and Phlegm Removing Prescription can redu ce fasting blood glucose, improve the insulin resistance, and prevent metabolic syndr ome. |
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