丹参酮ⅡA对子宫内膜异位症大鼠的干预作用及对TGF-β/SMADS信号通路的影响

Intervention Effect of TanshinoneⅡA on Endometriosis in Rats and Its Influence on TGF-β/SMADS Signaling Pathway

目的:探究丹参酮ⅡA对子宫内膜异位症(EMS)大鼠的干预作用,分析其作用机制。方法:采用自体子宫内膜移植术建立EMS大鼠模型,将大鼠随机分为假手术对照组、模型对照组、孕三烯酮0.5 mg/kg组、丹参酮ⅡA 30 mg/kg组、丹参酮ⅡA 30 mg/kg+SRI-01138110 mg/kg组、SRI-01138110 mg/kg组。治疗4 w后,以游标卡尺测量移植物体积,HE染色观察异位内膜病理变化,RT-qPCR法检测子宫内膜血管内皮生长因子(Vegf)、基质金属蛋白酶-9(Mmp9)、B细胞淋巴瘤-2基因(Bcl2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-9(Caspase9)mRNA表达,Western blot法检测子宫内膜转化生长因子-β1(TGF-β1)、SMAD2/3、p-SMAD2/3和SMAD7蛋白表达。结果:与假手术对照组比较,模型对照组大鼠异位子宫内膜体积显著增加,HE染色显示异位内膜上皮增生,间质细胞和腺体数目较多,血管丰富,子宫内膜异位组织Vegf、Mmp9、Bcl2 mRNA水平显著上调,Bax、Caspase9 mRNA水平显著下调,TGF-β1、p-SMAD2、p-SMAD3蛋白表达显著上调,SMAD7蛋白表达显著下调(P<0.01)。与模型对照组比较,丹参酮ⅡA 30 mg/kg组异位子宫内膜组织体积减小,HE染色显示异位内膜上皮增生和腺腔形成减少,血管形成减少。子宫内膜异位组织Vegf、Mmp9、Bcl2 mRNA表达下调,Bax、Caspase9 mRNA表达上调,TGF-β1、p-SMAD2、p-SMAD3蛋白表达下调,SMAD7蛋白表达上调(P<0.05或P<0.01)。TGF-β激活剂SRI-011381可抑制丹参酮ⅡA对EMS的改善作用,促进EMS发展,进一步激活TGF-β/SMADS信号通路(P<0.05)。结论:丹参酮ⅡA对大鼠EMS具有改善作用,其作用机制可能与抑制TGF-β/SMADS信号通路的激活有关。

Objective:To explore the intervention effects of tanshinoneⅡA on endometriosis(EMS)in rats and analyze its mechanism of action.Methods:After being modeled by autologous endometrial transplantation,the EMS rats were randomly divided into the sham operation group,model group,0.5 mg/kg gestrinone group,30 mg/kg tanshinoneⅡA group,30 mg/kg tanshinoneⅡA+10 mg/kg SRI-011381 group,and 10 mg/kg SRI-011381 group.After four weeks of treatment,the graft volume was measured by vernier caliper,and the pathological changes in ectopic endometrium were observed after HE staining.The mRNA expression levels of vascular endothelial growth factor(Vegf),matrix metalloproteinase-9(Mmp9),B-cell lymphoma-2(Bcl2),Bcl-2-associated X protein(Bax),and cysteine-dependent aspartate-directed protease-9(Caspase9)were detected by RT-qPCR,and the protein expression levels of transforming growth factor-β1(TGF-β1),SMAD2/3,p-SMAD2/3,and SMAD7 in endometrium by Western blotting.Results:Compared with the sham operation group,the model group exhibited significantly increased ectopic endometrial volume.HE staining revealed ectopic endometrial epithelial hyperplasia,a large number of mesenchymal cells and glands,and abundant blood vessels.The mRNA expression levels of Vegf,Mmp9 and Bcl2 and the protein expression levels of TGF-β1,p-SMAD2,and p-SMAD3 were significantly up-regulated,while the mRNA expression levels of Bax and Caspase9 and the SMAD7 protein expression were down-regulated(P<0.01).Compared with the model group,30 mg/kg tanshinoneⅡA reduced the ectopic endometrial volume,alleviated ectopic endometrial epithelial hyperplasia,glandular cavity formation,and angiopoiesis,down regulated the mRNA expression of Vegf,Mmp9,and Bcl2 as well as the protein expression of TGF-β1,p-SMAD2,and p-SMAD3,and up regulated the mRNA expression of Bax and Caspase9 and SMAD7 protein expression(P<0.05 or P<0.01).The TGF-βactivator SRI-011381 inhibited the improving effect of tanshinoneⅡA on EMS,thus promoting the development of EMS and further activating the TGF-β/SMADS signaling pathway(P<0.05).Conclusion:TanshinoneⅡA improves EMS in rats,which may be related to its inhibition of the activation of TGF-β/SMADS signaling pathway.