定坤丹通过microRNA-30d-5p靶向调控卵巢组织Smad 2治疗多囊卵巢综合征模型大鼠的机制研究  

Study on the mechanism of Dingkundan targeting and regulating Smad2 in ovarian tissue through microRNA-30d-5p to treat PCOS model rats

作　　者：陈兰[1] 杨波 罗立波 颜晓静[1] 王地均 蔡佳丽 尹春燕 CHEN Lan;YANG Bo;LUO Li-bo;YAN Xiao-jing;WANG Di-jun;CAI Jia-li;YINChun-ya(Changzhou Hospital of Traditional Chinese Medicine,Changzhou 213003,China)

机构地区：[1]常州市中医医院,江苏常州213003 [2]广誉远中药股份有限公司,山西太原030000

出　　处：《中国实用妇科与产科杂志》2024年第8期849-854,共6页Chinese Journal of Practical Gynecology and Obstetrics

基　　金：国家自然科学基金-青年科学基金(82104905);常州市孟河医派人用经验研究与转化重点实验室(CM20223006);南京中医药大学技术开发项目(2017005)。

摘　　要：目的通过观察定坤丹对多囊卵巢综合征(polycystic ovary syndrome,PCOS)模型大鼠卵巢组织形态学、血清性激素水平、转化生长因子β1(transforming growth factor-β1,TGF-β1)、Smad 2、Smad 3和microRNA-30d-5p表达的影响,进而探讨其通过microRNA-30d-5p靶向调控卵巢组织中Smad2治疗PCOS模型大鼠的作用机制。方法应用高、中、低剂量定坤丹[4.5、2.25、1.125 g/(kg·d)]干预脱氢表雄酮(dehydroepiandrosterone,DHEA)诱导的PCOS大鼠模型,采用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)、苏木精-伊红(hematoxylineosin,HE)染色、免疫组化、实时聚合酶链式反应(real time polymerase chain reaction,RT-PCR)及Western blot等方法检测各组大鼠卵巢组织形态学、血清性激素水平、microRNA-30d-5p的表达及TGF-β1、Smad 2、Smad 3 m RNA及蛋白的表达。结果与模型大鼠组相比较,定坤丹高、中、低剂量组卵巢分布有各级卵泡,可见较多黄体;雌二醇(estradiol,E_(2))、黄体生成素(luteinizing hormone,LH)及睾酮(testosterone,T)均下降,卵泡刺激素(follicle stimulating hormone,FSH)则升高,且差异均具有统计学意义;microRNA-30d-5p的表达升高,且差异均具有统计学意义;TGF-β1、Smad 2及Smad 3的mRNA及蛋白表达均下降,且差异均具有统计学意义。结论DHEA诱导的PCOS大鼠模型是研究PCOS排卵障碍较为理想的动物模型。定坤丹对PCOS大鼠有一定治疗作用,其机制可能是通过microRNA-30d-5p靶向调控卵巢组织中Smad 2的表达水平来实现。Objective To observe the effects of Dingkundan on ovarian tissue morphology,serum sex hormone levels,transforming growth factor-β1(TGF-β1),Smad 2,Smad 3,and microRNA-30d-5p expression in polycystic ovary syn-drome(PCOS)model rats,and to further explore the mechanism of Dingkundan's targeted regulation of Smad 2 in ovari-an tissue through microRNA-3Od-5p in treating PCOS model rats.Methods DHEA-induced PCOS rat models were treated with low[1.125 g/(kg·d)],medium[2.25 g/(kg·d)]and high[4.5 g/(kg•d)]doses of Dingkundan.Enzyme linked immunosorbent assay(ELISA),hematoxylin-eosin(HE)staining,immunohistochemistry,real time polymerase chain reaction(RT-PCR)and Western blot were used to detect the ovarian histomorphology,serum sex hormone levels,microRNA-30d-5p expressions,as well as protein and mRNA expressions of TGF-β1,Smad 2 and Smad 3.Results Compared with model group,all Dingkundan treatment groups significantly promoted follicles of all levels and corpus luteum in the ovaries,downregulated the levels of estradiol(E_(2)),luteinizing hormone(LH)and testosterone(T)while in-creasing follicle stimulating hormone(FSH)level,the differences being statistically significant;both protein and mRNA expressions of TCF-β1,Smad 2 and Smad 3 decreased while microRNA-30d-5p expressions increased,both differences being statistically significant.Conclusions The DHEA-induced PCOS rat model is an ideal animal model for studying ovulation disorders in PCOS.Dingkundan has a certain therapeutic effect on PCOS rats,and its mechanism may be to target and regulate the expression level of Smad 2 in ovarian tissue through microRNA-30d-5p.

关 键 词：多囊卵巢综合征 定坤丹 转化生长因子β1 Smad 2 Smad 3 microRNA-30d-5p 

分 类 号：R711.75[医药卫生—妇产科学]