基于TGF-β1/Smad通路探究萝卜硫素对EMS的作用

Exploring the effect of sulforaphane on EMS based on TGF-β1/Smad pathway

目的 探究萝卜硫素对子宫内膜异位症(EMS)大鼠的作用及其作用机制。方法 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组、模型组、萝卜硫素组、萝卜硫素+SRI-011381(TGF-β激活剂)组和SRI-011381组,每组10只。测定大鼠子宫内膜异位组织体积,HE染色观察组织病理变化,ELISA法检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL^(-1)β)和白细胞介素-6(IL-6)水平,Western blot检测异位内膜组织血管内皮生长因子(VEGF)、基质金属蛋白酶-9(MMP-9)及转化生长因子-β1(TGF-β1)、Smad2/3、p-Smad2/3和Smad7蛋白表达水平。结果 模型组大鼠异位子宫内膜组织体积大于假手术组,异位病灶腺上皮细胞和基质细胞密集,腺腔完整,TNF-α、IL^(-1)β、IL-6含量和VEGF、MMP-9蛋白表达水平显著升高,TGF-β1、p-Smad2/3蛋白表达显著升高,Smad7蛋白表达显著降低(P<0.05)。与模型组比较,萝卜硫素可显著缩小异位子宫内膜组织的体积,使腺上皮细胞胞核体积缩小,排列稀疏,腺腔缩小,内膜腺上皮层变薄,呈萎缩性改变,TNF-α、IL^(-1)β、IL-6含量和VEGF、MMP-9蛋白表达水平降低,TGF-β1、p-Smad2/3蛋白表达减少,Smad7蛋白表达增加(P<0.05),TGF-β激活剂SRI-011381可抑制萝卜硫素对EMS的作用,促进EMS发展,进一步激活TGF-β1/Smad信号通路(P<0.05)。结论 萝卜硫素对大鼠EMS具有治疗作用,其机制可能与调节TGF-β1/Smad通路有关。

Objective To explore the effect of sulforaphane on endometriosis(EMS) in rats and its mechanism.Methods Autologous endometrial transplantation was used to establish EMS rat models. The rats were randomly divided into sham operation group, model group, sulforaphane group, sulforaphane+SRI-011381(TGF-β activator) group, SRI-011381 group, each with 10 rats. The volume of rat endometriosis tissues was measured, and HE staining was used to observe histopathological changes. ELISA kits were used to detect the levels of serum tumor necrosis factor-α(TNF-α), interleukin-1β(IL^(-1)β) and interleukin-6(IL-6). Western blot was used to detect the protein expression levels of VEGF, MMP-9 and TGF-β1, Smad2/3, p-Smad2/3 and Smad7 in ectopic endometrial tissues.Results Compared with sham operation group, the volume of ectopic endometrial tissues in model group increased significantly, the ectopic lesions were densely packed with glandular epithelial cells and stromal cells, the glandular cavity was intact, and the contents of TNF-α, IL^(-1)β, IL-6 and VEGF, MMP-9 protein expression levels increased, TGF-β1, p-Smad2/3 protein expression increased significantly, and Smad7 protein expression decreased significantly(P<0.05). Compared with model group, sulforaphane significantly reduced the volume of ectopic endometrial tissues, reduced the nucleus volume of glandular epithelial cells, sparsely arranged, reduced glandular cavity, and thinned endometrial glandular epithelial layer, showing atrophy. The levels of TNF-α, IL^(-1)β, IL-6, VEGF and MMP-9 protein expression decreased, TGF-β1, p-Smad2/3 protein expression decreased, and Smad7 protein expression increased(P<0.05). The TGF-β activator SRI-011381 inhibited the effect of sulforaphane on EMS, promoted the development of EMS, and further activated TGF-β1/Smad signaling pathway(P<0.05).Conclusion Sulforaphane has a therapeutic effect on EMS in rats, and its mechanism may be related to the regulation of TGF-β1/Smad pathway.