Based On PI3K/AKT/mTOR Torsignal Pathway,the Experimental Study Of Guizhi Fuling Pill Regulating Ovulation Disorder In PCOS-IR Rats

Objective: In order to make more scientific and repeatable results for the formal experiment,the pre experiment in the early stage explored the modeling method,and considered letrozole combined with high-fat emulsion feeding for 35 days as the modeling method for the formal experiment.The main purpose of this experiment is to establish polycystic ovary syndrome with insulin resistance(pcos-ir)model rats by letrozole combined with high-fat emulsion.The model rats were given Guizhi Fuling pill to observe the effect of Guizhi Fuling Pill on ovulation,endocrine and metabolism disorders of pcos-ir model rats,observe the change of PI3K/Akt/mTORsignal pathway,and analyze pcos-ir and PI3K/Ak K/mTOR The relationship between the R signal pathway and PCOSwas discussed.Therefore,the purpose of this study is to explore the possible therapeutic value of Guizhi Fuling pill in reversing the abnormal PCOSof letrozole model rats,and to compare it with metformin.Method: Forty two Sprague Dawley female healthy rats,6 weeks old,weighing 150 ± 10 g,were randomly divided into blank control group,model group,Guizhi Fuling pill low-dose group,Guizhi Fuling pill middle dose group,Guizhi Fuling pill high-dose group and metformin group.All rats were fed adaptively for 4 days.The blank control group was given 10 ml / kg of normal saline by gavage daily for 35 days.The model group was given high-fat emulsion(lard + cholesterol,lard: cholesterol = 10:3)and letrozole(1mg / kg · d)carboxymethylcellulose sodium(CMC Na)solution once a day for 35 days.The low-dose group was given high-fat emulsion and letrozole daily At the same time,Guizhi Fuling pill suspension(10ml / 1kg for each rat in normal saline configuration)equivalent to 0.31(g / kg · d)dose of crude drug was taken orally for 35 consecutive days;the middle dose group of Guizhi Fuling pill was given high-fat emulsion and letrozole orally at the same time,Guizhi Fuling pill suspension(10ml / 1kg for each rat in normal saline configuration)equivalent to 0.62(g / kg · d)dose of crude drug was taken orally for 35 consecutive days Days;Guizhi Fuling pill high dose group was given high-fat emulsion and letrozole at the same time,which was equivalent to 1.24(g / kg · d)dose of Guizhi Fuling pill suspension(with normal saline,10 ml / 1kg for each rat)for 35 days;metformin group was given high-fat emulsion and letrozole at the same time,270 mg / kg · D dose of metformin suspension for 35 days.The content of FSH,LH,t,E2 and fins were detected by enzyme-linked immunosorbent assay(ELISA).The protein expression of p-pi3 k,p-Akt and p-mTOR in rat ovarian granulosa cells was detected by Western blot.Result:(1)Judgment of rat PCOS-IR modelCompared with the normal control group,the rats in the model group have a basically disordered estrous cycle.The weight of the model group rats is significantly greater than that of the normal control group,and the difference is statistically significant(P <0.05).H & E staining of ovarian tissue in the model group showed an increase in the number of cystic follicles and atretic follicles.Compared with the normal control group,fasting insulin levels in the model group increased,fasting blood glucose levels increased,and insulin resistance index increased.-IR rat model.(2)Effect of Guizhi Fuling Pill on serum T,E2,FSH,LH levels in ratsCompared with the model group,the serum levels of E2 and FSH increased,the levels of LH and T decreased,and the ratio of LH / FSH decreased in the Guizhi Fuling Pill group and the metformin group.The difference was statistically significant(P <0.05).(3)Effect of Guizhi Fuling Pill on FINS,FBG,HOMA-IR results in rat serumCompared with the model group,the serum FINS level,the fasting blood glucose content,and the insulin resistance index of the rats in the Guizhi Fuling Pill group and the metformin group decreased,and the differences were statistically significant(P <0.05).(4)Effect of Guizhi Fuling Pill on weight gain in ratsCompared with the model group,the weight gain of Guizhi Fuling Pill rats in the high,medium and low dose groups and metformin group decreased,and the difference was statistically significant(P <0.05).(5)Effect of Guizhi Fuling Pill on follicles and corpus luteum of ovarian tissue in ratsCompared with the model group,the number of follicles and corpus luteum in the ovaries of Guizhi Fuling Pill high-and medium-dose groups increased,polycystic ovarian changes were reduced,and the number of cystic dilated follicles and atretic follicles was reduced in the model group.The difference was statistically significant(P <0.05).(6)Effect of Guizhi Fuling Pill on p-PI3 k,p-AKT,and p-mTOR levels in rat ovarian granulosa cellsCompared with the model group,the expressions of p-PI3 k,p-AKT,and p-mTOR in ovarian granulosa cells of rats in Guizhi Fuling Pill high and medium dose group and metformin group were up-regulated,and the differences were statistically significant(P <0.05).Conclusion:(1)Letrozole combined with high-fat emulsion can successfully construct a rat model of polycystic ovary syndrome with insulin resistance(PCOS-IR).(2)PCOS-IR rats showed weight gain,reduced levels of ovarian follicles and corpus luteum,cystic dilated follicles,and increased atretic follicles.The mechanism may be insulin resistance in the PCOS-IR model rats,reducing PI3 k / AKT.Transduction of the / mTOR signaling pathway.(3)Guizhi Fuling Pill can reduce the weight of rats,increase the number of follicles and corpus luteum at various levels in the rat’s ovary,reduce the appearance of cystic dilated follicles,reduce atretic follicles,reduce androgen levels,LH / FSH ratio,and possibly It is to improve the insulin sensitivity of PCOS-IR rats,improve the insulin resistance of PCOS-IR model rats,further activate the PI3 K / Akt / mTOR signaling pathway,and improve the condition of PCOS ovulation disorders.