摘要
目的:研究归术益坤方治疗多囊卵巢综合征(PCOS)的作用机制。方法:通过体外培养大鼠卵巢颗粒细胞,以丙酸睾丸酮(TP)、自噬诱导剂C2-神经酰胺作用颗粒细胞24h,建立大鼠PCOS颗粒细胞的自噬模型;实验分为空白组,模型组,归术益坤方低、中、高剂量组及自噬阻断剂3MA组。通过Western blot法检测各组颗粒细胞中LC3Ⅱ、p53、p-m TOR、p-AMPK的表达。结果:与空白组比较,模型组LC3Ⅱ的表达显著增加(P<0.01);与模型组比较,归术益坤方高、中、低剂量组表达降低(P<0.01)。与空白组比较,模型组p53、p-m TOR显著降低(P<0.01),p-AMPK升高(P<0.01);与模型组比较,高剂量归术益坤方可以上调p53和p-m TOR的表达(P<0.01,P<0.05);高、中剂量归术益坤方可下调p-AMPK的表达(P<0.01)。结论:归术益坤方可以通过调控p53/AMPK信号通路来抑制自噬,这可能是归术益坤方治疗PCOS的新机制。
Objective: To explore the molecular mechanism of Guizhu Yikun Formula(GZYKF) in treating polycystic ovarian syndrome(PCOS). Methods: Granulosa cells(GCs) in PCOS rats were cultured and treated with testosterone propionate(TP) and C2-ceramide for 24 h to establish PCOS cell model in vitro. GCs were divided into blank group, model group, 3 MA group, GZYKF high, middle and low dose groups. The expression of LC3 II, p53, p-m TOR, and p-AMPK were observed by Western blot. Results: Compared with blank group, the expression of LC3 II was significantly increased in model group(P<0.01).Compared with model group, the expression of LC3 II in the high, middle, low dose GZYKF group decreased(P<0.01). Compared with the blank group, the expression of p53, p-m TOR in the model group decreased(P<0.01), the expression of p-AMPK increased(P<0.01). Compared with model group, high dose GZYKF could increase the expression of p53 and p-m TOR(P<0.01, P<0.05), and high, middle dose GZYKF could decrease the expression of p-AMPK(P<0.01). Conclusion: The effects of GZYKF on mediating the p53/AMPK pathway could inhibit GC autophagy which suggesting a possible novel mechanism by GZYKF's treatment of PCOS.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2017年第11期5074-5077,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81202720)
北京中医药大学自主课题(No.2015-JYB-XS181)~~