Objective:To observe oftonifying kidney and resolving phlegm(TKRP)method on the obese PCOS with glucose and lipid metabolism disorder in rats and study the regulation of APN/p38 MAPK signal pathway and the expression of genes related to glycolipid metabolism in rat ovarian granulosa cells(GC).To explore the effect and mechanism of Chinese medicine TKRP method to improve the obese PCOS.Method: The healthy female rats aged 7~8 weeks with normal estrous cycle were selected and randomly divided into blank control group and obese PCOS by model group ratio 1:5 according to experimental requirements.The obese PCOS model was prepared by intragastric administration of letrozole(LE)combined with high fat suspension for 28 days.PCOS model animals were randomly divided into model control group,high dose group of TKRP,TKRP middle dose group,low dose group of TKRP and Diane-35 group and each dose group of Chinese medicine dose group and Diane-35 group were continuous administration of medicine for 3 weeks.Serum APN content was by ELISA method;enzyme method was used to detect the expression of serum FBG;immunohistochemical method was used to detect the expression of Adipo R2,PPAR-?,FATP1 in the GC of the ovarian;relative expression of PPAR-?m RNA,PPAR-?m RNA,p38 MAPK m RNA in the ovarian GC by realtime PCR.Result:Compared with the blank control group,model group rats increased in body mass growth rate with differences were statistically significant(P<0.05),compared with the blank control group,model group rats estrous cycle disorder,HE staining of the model group showed that the number of cystic follicular and atresia follicles increased,determination of successful model in obese PCOS rats;Compared with the model control group,TKRP high,middle and low dose group rats the number of primary follicles,secondary follicles,mature follicles and corpus luteum increased,the number of cystic follicular and atresia follicles decreased in the ovary;Compared with the model control group,TKRP high,middle dose group and Diane-35 group rats the content of FSH and E2 increased and the content of LH and T decreased in serum with differences were statistically were significant(P<0.05);Compared with the model control group,TKRP high,middle dose group and Diane-35 group rats decreased in body mass growth rate with differences were statistically significant(P <0.05);Compared with the model control group,TKRP high,middle dose group and Diane-35 group rats the expression content of APN in serum was increased with differences were statistically significant(P<0.05);Compared with the model control group,TKRP high,middle dose group and Diane-35 group rats increased of Adipo R2 expression in ovarian GC with differences were statistically significant(P <0.05);Compared with the model control group,TKRP high,middle dose group and Diane-35 group rats increased of FATP1 expression in ovarian GC with differences were statistically significant(P<0.05);Compared with the model control group,TKRP high,middle dose group and Diane-35 group rats increased of PPAR-? expression in the ovarian GC with differences were statistically significant(P < 0.05);Compared with the model control group,TKRP high,middle dose group and Diane-35 group rats decreased of FBG in serum with differences were statistically significant(P <0.05);Compared with the model control group,TKRP high,middle dose group and Diane-35 group rats increased of p38 MAPK m RNA expression in the ovarian GC with differences were statistically significant(P<0.05);Compared with the model control group,TKRP high,middle,low dose group and Diane-35 group rats increased of PPAR-?/ ? m RNA expression in the ovarian GC with differences were statistically significant(P<0.05).Conclusion:(1)Obese PCOS rats showed obvious disorder of glucose and lipid metabolism and the mechanism may be related to abnormal APN/p38 MAPK signal transduction.(2)TKRP method may be improved the obese PCOS with glycolipid metabolic disorder by the APN/p38 MAPK signal pathway. |