基于micRNA表达谱分析加味桂枝茯苓丸干预大鼠子宫肌瘤的分子机制

Molecular Mechanism of Modified Guizhi Fulingwan Intervention on Uterine Fibroids in Rats Based on micRNA Expression Profiling

目的:探讨加味桂枝茯苓丸干预子宫肌瘤大鼠的分子机制。方法:将72只无特定病原体(SPF)雌性成年SD大鼠随机分为模型组、正常组、预防给药组,模型组及预防性给药组采用雌孕激素负荷法建立子宫肌瘤大鼠模型。造模成功后将模型组大鼠随机分为米非司酮组,加味桂枝茯苓丸大、小剂量组。所有大鼠分别灌胃,正常组和模型组灌胃等量生理盐水,米非司酮组(灌胃米非司酮1.04 mg·kg^(-1)),加味桂枝茯苓丸大、小剂量组(分别灌胃加味桂枝茯苓丸9.18,4.59 g·kg^(-1))、预防给药组(灌胃加味桂枝茯苓丸4.59 g·kg^(-1)),1次/d,连续28 d。采用苏木精-伊红(HE)染色观察子宫形态学变化,采用互补干扰RNA(micRNA)基因芯片检测子宫micRNA基因表达谱,通过生物信息学技术筛选出有差异表达的micRNA,通过基因功能富集预测加味桂枝茯苓丸干预子宫肌瘤模型大鼠可能的信号通路。结果:与正常组比较,模型组microRNA上调1个,下调9个。与模型组比较加味桂枝茯苓丸大剂量组上调2个,下调1个;预防给药组上调9个,下调2个。经过基因功能富集分析,4条信号通路与子宫肌瘤密切相关,分别是丝裂原活化蛋白激酶(MAPK)信号通路,Wnt信号通路,哺乳动物雷帕霉素靶蛋白(mTOR)信号通路,血管内皮细胞生长因子(VEGF)信号通路。结论:加味桂枝茯苓丸影响了雌孕激素诱导子宫肌瘤模型大鼠micRNA的表达谱,提示桂枝茯苓丸治疗子宫肌瘤可能涉及到多种生物学过程信号传导及基因调控,是一个复杂的病理生理过程。

Objective:To explore the molecular mechanism of modified Guizhi Fulingwan in rats with uterine fibroids. Method: Seventy-two female adult SD rats of SPF grade were randomly divided into a model group,a normal group,and a preventive administration group. The model group and preventive administration group were established by estrogen and progestin loading method. After successful modeling,the rats in the model group were randomly divided into a western medicine group(mifepristone),the high-dose traditional Chinese medicine(TCM)group,and a low-dose TCM group. All the rats were dosing as required once a day for28 consecutive days. Hematoxylin-eosin(HE)staining was used to observe the morphological changes of the uterus. The micRNA gene chip was used to detect the expression profile of uterine micRNA gene. Differential expressions of micRNA were screened by bioinformatics methods. Gene function enrichment was used to predict the possible signaling pathways in rats with uterine fibroids by modified Guizhi Fulingwan. Result: Compared with the normal group,microRNA of the model group was 1 up-regulated and 9 down-regulated. Compared with the model group,microRNA of the high-dose group of TCM group was 2 up-regulated and 1 down-regulated,in the preventive administration group,9 was up-regulated and 2 was down-regulated. Gene function enrichment analysis indicated that four signaling pathways were closely related to uterine fibroids. They were mitogenactivated protein kinase(MAPK)signaling pathway,Wnt signaling pathway,mammalian rapamycin target protein(mTOR)signaling pathway and vascular endothelial cell growth factor(VEGF)signaling pathway.Conclusion: Modified Guizhi Fulingwan affected the expression profile of micRNA in rat model of uterine fibroids induced by estrogen and progesterone,suggesting that modified Guizhi Fulingwan may involve in a variety of biological processes such as signal transduction and gene regulation in the treatment of uterine fibroids.