苍附导痰汤对来曲唑致多囊卵巢综合征模型大鼠卵巢形态和卵泡发育的影响

Effect of Cangfu Daotan Decoction on Ovarian Morphology and Follicular Development in Letrozole Induced Polycystic Ovary Syndrome Model Rats

目的研究苍附导痰汤(Cangfu Daotan decoction,CFDTD)对来曲唑致多囊卵巢综合征(polycystic ovary syndrome,PCOS)模型大鼠卵巢形态和卵泡发育的影响。方法SD大鼠给予来曲唑制备PCOS模型,并行阴道涂片观察动情周期,随机分为PCOS组,CFDTD 15,30 g·kg^(-1)·d^(-1)组、二甲双胍(metformin,MH,50 mg·kg^(-1)·d^(-1))组,各10只,分别给予等量生理盐水及相应药物灌胃,并以高脂饲料喂养;同时设置对照组,等量生理盐水灌胃、普通饲料喂养,共给药4周。给药结束后腹主动脉采血并收集卵巢组织,检测各组大鼠空腹血糖(fasting blood glucose,FPG)、空腹胰岛素(fasting insulin,FINS)、卵巢质量、卵巢直径、卵巢脏器指数、血清促黄体生成素(luteinizinghormone,LH)、促卵泡生长激素(follicle stimulatinghormone,FSH)、睾酮(testosterone,T)水平。同时采用HE染色、Tunel染色观察卵巢组织形态、细胞凋亡情况。Western blotting检测卵巢组织cleaved caspase 3、cleaved caspase 9、Bcl-2蛋白表达。结果PCOS大鼠无规则动情周期。与PCOS组相比,CFDTD 15 g·kg^(-1)·d^(-1)组左卵巢质量、血清LH、T水平显著降低(P<0.05),CFDTD 30 g·kg^(-1)·d^(-1)组与MH组大鼠FPG、FINS、胰岛素抵抗指数水平、体质量、左右卵巢质量、卵巢直径、卵巢脏器指数、血清LH、T水平显著降低(P<0.05或P<0.01)。HE染色、Tunel染色结果显示,与PCOS组相比,给药组卵巢组织形态改善、阳性细胞凋亡数减少,改善程度从大到小为MH组、CFDTD 30 g·kg^(-1)·d^(-1)组、CFDTD 15 g·kg^(-1)·d^(-1)组。Western blotting检测结果显示给药组可下调cleaved caspase 3、cleaved caspase 9蛋白表达,上调Bcl-2蛋白表达。结论CFDTD可通过抑制颗粒细胞凋亡、改善胰岛素抵抗和生殖激素水平来发挥改善PCOS大鼠的卵巢形态和卵泡发育的治疗作用。

OBJECTIVE To study the effect of Cangfu Daotan decoction(CFDTD)on ovarian morphology and follicular development in letrozole induced polycystic ovary syndrome(PCOS)rats.METHODS SD rats were given letrozole to establish PCOS model,and the estrus cycle was observed by vaginal smear.SD rats were randomly divided into PCOS group,CFDTD 15,30 g·kg^(-1)·d^(-1)groups,and metformin(MH,50 g·kg^(-1)·d^(-1))group with 10 rats in each group.The same amount of normal saline and corresponding were treated by gavage respectively,and fed with high-fat diet,at the same time,the control group was given the same amount of normal saline and ordinary feed,all rats were treated for 4 weeks.After administration,blood was collected from abdominal aorta and ovarian tissue was collected,and fasting blood glucose(FPG),fasting insulin(FINS),ovarian weight,ovarian diameter,ovarian organ index,serum luteinizing hormone(LH),follicle stimulating hormone(FSH)and testosterone(T)levels were detected.Meanwhile,HE staining and Tunel staining were used to observe the morphology of ovarian tissue and apoptosis.The protein expression of cleaved caspase 3,cleaved caspase 9 and Bcl-2 were detected by Western blotting.RESULTS There was no regular estrus cycle in PCOS rats.Compared with the PCOS group,the left ovarian mass,serum LH and T levels in CFDTD 15 g·kg^(-1)·d^(-1)group were significantly decreased(P<0.05),in the CFDTD 30g·kg^(-1)·d^(-1)group and MH group,the levels of FPG,FINS,homeostasis model of assessment of insulin resistance(HOMA-IR),body weight,diameter of left and right ovaries,ovarian organ index,serum LH and T levels were significantly decreased(P<0.05or P<0.01).The results of HE staining and Tunel staining showed that compared with the PCOS group,the morphology of ovarian tissue was improved and the number of apoptosis positive cells in the treatment group was decreased.The improvement degree was as follows:MH group,CFDTD 30 g·kg^(-1)·d^(-1)group,CFDTD 15 g·kg^(-1)·d^(-1)group.Western blotting analysis showed that the expression of cleaved caspase 3 and cleaved caspase 9 protein was down-regulated and Bcl-2 protein expression was up-regulated in the treatment group.CONCLUSION CFDTD can improve the ovarian morphology and follicular development of PCOS rats by inhibiting the apoptosis of granulosa cells,improving insulin resistance and reproductive hormone levels.