基于TGF-β1/Smad通路探究补肾化瘀方对宫腔粘连大鼠子宫内膜纤维化的作用及机制  

Study on the Effect and Mechanism of Bushen Huayu Recipe(补肾化瘀方)on Endometrial Fibrosis in Rats with Uterine Adhesion Based on TGF-β1/Smad Pathway

作　　者：李姣[1] 林洁[1] 易星星[1] 邱冉冉[1] LI Jiao;LIN Jie;YI Xing-xing;QIU Ran-ran(The First Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410007,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《中医药导报》2021年第11期7-11,17,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省自然科学基金项目(2017JJ3242)。

摘　　要：目的:观察补肾化瘀方对宫腔粘连(IUA)大鼠子宫内膜纤维化的改善作用,探究其作用机制。方法:采取机械+感染双重损伤法建立IUA大鼠模型,将造模大鼠随机分为模型组、补肾化瘀方组、补肾化瘀方+SRI-011381组、SRI-011381组,每组15只。另取15只大鼠作为假手术组。药物治疗21 d后,观察大鼠子宫形态,HE染色观察子宫组织病理变化,计数子宫内膜腺体数量,Masson染色观察纤维化情况,ELISA试剂盒检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)的水平,qRT-PCR检测子宫组织转化生长因子-β1(TGF-β1)mRNA、Smad3 mRNA、Smad4 mRNA和Smad7 mRNA表达水平,Western blotting法测定TGF-β1、Smad3、Smad4、Smad7、p-Smad3、p-Smad4、p-Smad7水平。结果:与假手术组比较,模型组大鼠子宫组织增粗,腺体数量减少,纤维化面积增加,血清炎症因子水平升高,TGF-β1 mRNA、Smad3 mRNA、Smad4 mRNA表达及Smad3、Smad4磷酸化增加,Smad7 mRNA表达及磷酸化减少,差异均有统计学意义(P<0.05)。与模型组比较,补肾化瘀方组大鼠子宫腺体数量增加,纤维化面积减少,血清炎症因子水平降低,TGF-β1 mRNA、Smad3 mRNA、Smad4 mRNA表达及Smad3、Smad4磷酸化减少,Smad7 mRNA表达及磷酸化增加,差异均有统计学意义(P<0.05)。SRI-011381可抑制补肾化瘀方的作用,促进纤维化,进一步激活TGF-β1/Smad通路(P<0.05)。结论:补肾化瘀方对IUA大鼠子宫内膜纤维化有改善作用,其机制可能与调节TGF-β1/Smad通路有关。Objective:To observe the effect of Bushen Huayu Recipe on the improvement of intrauterine adhesion(IUA)endometrial fibrosis in rats and explore its mechanism of action.Methods:The mechanical and infection injury method was adopted to establish an IUA rat model.The rats were randomly divided into sham operation group,model group,Bushen Huayu Recipe group,Bushen Huayu Recipe+SRI-011381(TGF-βactivator)group,and SRI-011381 group,with 15 rats in each group.After 21 days of treatment,the morphology of the rat’s uterus was observed.HE staining was used to observe pathological changes in uterine tissues and count the number of endometrial glands.Masson staining was used to observe uterine fibrosis.ELISA kit was used to detect serum tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)levels.qRT-PCR was used to detect the mRNA expression levels of transforming growth factor-β1(TGF-β1),Smad3,Smad4 and Smad7 in uterine tissues.The levels of TGF-β1,Smad3,Smad4,Smad7,p-Smad3,p-Smad4,and p-Smad7 were determined by Western blotting.Results:Compared with sham operation group,the uterine tissue of model group became thicker,the number of glands decreased,the area of fibrosis increased,the level of serum inflammatory factors increased,TGF-β1,Smad3,Smad4 mRNA expression and Smad3,Smad4 phosphorylation increased,and Smad7 mRNA expression and phosphorylation decreased.The differences were statistically significant(P<0.05).Compared with model group,the number of uterine glands in the Bushen Huayu Recipe group increased,the area of fibrosis decreased,the level of serum inflammatory factors decreased,the expression of TGF-β1,Smad3,Smad4 mRNA and the phosphorylation of Smad3,Smad4 decreased,and the expression of Smad7 mRNA was increased.The differences were statistically significant(P<0.05).SRI-011381 inhibited the effect of Bushen Huayu Recipe,promoted fibrosis,and further activated TGF-β1/Smad pathway(P<0.05).Conclusion:Bushen Huayu Recipe can improve endometrial fibrosis in IUA rats,and its mechanism

关 键 词：宫腔粘连 子宫内膜纤维化 补肾化瘀方 TGF-β1/Smad通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]