补肾化痰法经Sphk1通路调控肥胖PCOS雌鼠子宫内膜容受性的研究  

Effects of Bushen Huatan method on endometrial receptivity of obese PCOS female rats via SphK1 pathway

作　　者：陈美佳 徐晓娟[1] 黄映红[1] 田雪梅 张淼 熊倩 CHEN Mei-jia;XU Xiao-juan;HUANG Ying-hong;TIAN Xue-mei;ZHANG Miao;XIONG Qian(Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China)

机构地区：[1]成都中医药大学,成都610072

出　　处：《中国中医基础医学杂志》2020年第10期1488-1492,1510,共6页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金资助项目(81470199)-经APN信号通路探讨补肾化痰法调控肥胖型PCOS卵泡发育机制研究;四川省科技厅应用基础项目(2017JY0052)-补肾化痰法经Sphkl/SIP-PI3K/Akt信号通路调控肥胖型PCOS子宫内膜容受性的机制研究。

摘　　要：目的:观察补肾化痰法调控肥胖多囊卵巢综合征(PCOS)雌鼠子宫内膜容受性(ER)的作用机制。方法:采用7~8周龄SD雌鼠,来曲唑加高脂乳剂建立肥胖PCOS模型以补肾化痰法治疗,结合阴道脱落细胞涂片、体质量、血清性激素和子宫形态学进行模型验证。取子宫内膜进行苏木精-伊红(HE)染色,ELISA法检测血清雌二醇(E 2)、孕激素(PROG)及睾酮(T)的表达,免疫组织化学法检测孕激素受体(PR)、雌二醇受体(E 2R)的表达水平,RT-PCR法检测雌鼠子宫鞘氨醇激酶1(Sphk1)、1型1-磷酸鞘氨醇受体(S1Pr1)、蛋白激酶(Akt)、细胞周期蛋白1(CyclinD1)mRNA的表达。结果:与空白组比较,模型组动情周期紊乱,体质量显著增加,腺体个数明显减少,子宫内膜发育受损,血清T浓度及子宫内膜PR、E 2R表达升高,血清PROG、E 2浓度及子宫内膜Sphk1mRNA、S1Pr1mRNA、AktmRNA、CyclinD1mRNA表达显著降低;与模型组比较,中药高剂量组体质量增长得到抑制,腺体个数明显增加,子宫内膜发育改善,血清PROG、E 2浓度及子宫内膜Sphk1mRNA、S1Pr1mRNA、AktmRNA、CyclinD1mRNA表达升高,血清T浓度及子宫内膜PR、E 2R表达降低。结论:补肾化痰法可能通过纠正肥胖PCOS雌鼠生殖性激素紊乱,调控PR、E 2R,激活Sphk1信号通路,促进子宫内膜血管生成,从而发挥改善子宫内膜容受性的作用。Objective:To observe the mechanism of kidney-nourishing and phlegm-removing method(KNPR)in regulating endometrial receptivity in obese polycystic ovary syndrome female rats.Methods:We chose 7-8 weeks old female SD rats and treated them with the method of KNPR after modeling by Letrozole and high-fat and cholesterol.The models were verified by the smears of vaginal exfoliated cells,rats weight,the serum sex hormone levels and the morphological changes of uterine.The uterine tissues were taken out for HE staining,the serum level of E 2,PROG and T were detected by Elisa method,the expressions of PR and E 2R were detected by immunohistochemical staining method,The mRNA expressions of Sphk1,S1Pr1,Akt and CyclinD1 were detected by qPCR method.Results:Compared with the blank control group,in the model control group,The estrous cycle was disorder,the rats weight was significantly increased,the number of glands were significantly decreased,and the endometrial development was impaired,the T level was significantly increased,the PROG and E 2 levels were decreased,the expressions of PR and E 2R in the uterine tissue were increased,the mRNA expresinsions of Sphk1,S1Pr1,Akt and CyclinD1 were significantly decreased;Compared with the model control group,in the traditional Chinese medicine high dose group,the growth of the rats weight was suppressed,the number of glands were significantly increased,and the endometrial development was improved,tthe T level was decreased,the PROG and E 2 levels were increased,the expressions of PR and E 2R in the uterine tissue were decreased,the mRNA expresinsions of Sphk1,S1Pr1,Akt and CyclinD1 were increased.Conclusion:The method of KNPR may improve the ER of obese PCOS female rats by correcting the serum sex hormone disorder levels,regulating the expressions of PR and E 2R,activating Sphk1 signaling pathway,and promoting endometrial angiogenesis.

关 键 词：补肾化痰法 Sphk1通路 肥胖PCOS 子宫内膜容受性 

分 类 号：R285.5[医药卫生—中药学]