补肾益精法对系统性硬皮病血管内皮间质转化的影响  

Effects of Bushen Yijing Therapy on Endothelial to Mesenchymal Transition in Systemic Scleroderma

作　　者：邱路萍 田永贞 张佳林[1] 李东海[1,3] 宋文英[1,3] 齐庆[1,3] 

机构地区：[1]广州中医药大学第一临床医学院,广东广州510405 [2]广东省深圳市中医院,广东深圳518048 [3]广州中医药大学第一附属医院皮肤科,广东广州510405

出　　处：《上海中医药大学学报》2017年第1期63-68,共6页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81573976);广东省自然科学基金资助项目(2015A030313341);广东省科学技术厅项目(2014A020221029);广州中医药大学青年英才基金项目(20120103)

摘　　要：目的:探讨补肾益精法中药复方对系统性硬皮病血管内皮细胞内皮间质转化(endothelial to mesenchymal transition,End MT)的影响及机制。方法:采用博莱霉素建立硬皮病小鼠模型,造模同时补肾益精方软皮汤2号方(中药复方)进行干预,4周后HE染色观察小鼠皮肤组织病理、免疫组化法检测α-SMA表达水平;体外构建TGF-β1诱导的血管内皮细胞End MT模型,用补肾益精法含药血清处理48 h后,采用细胞免疫荧光方法检测血管内皮细胞α-SMA表达,Western Blot方法检测血管内皮细胞CD31、FSP1、Snail-1表达水平。结果:HE染色结果显示,模型组小鼠皮肤纤维化区域中,胶原纤维明显增多、胶原间隙变窄、毛细血管明显少于正常皮肤组织;与模型组比较,中药复方组小鼠皮肤胶原纤维较少、间隙较宽、血管数量较多;免疫组化染色检测结果显示,模型组小鼠皮肤组织中血管内皮细胞α-SMA表达高于正常对照组,中药复方组α-SMA阳性表达明显减少。免疫荧光检测结果显示,经TGF-β1诱导成功建立End MT模型,End MT模型组细胞α-SMA高表达,内皮细胞发生间质转化,经含药血清处理后,α-SMA表达明显减少。Western Blot检测结果显示,End MT模型组中Snail-1、FSP1均明显高于对照血清组,CD31明显低于对照血清组;含药血清处理组Snail-1、FSP1表达明显低于End MT模型组,CD31表达高于End MT模型组(P<0.05)。结论:补肾益精法对系统性硬皮病血管的保护作用,可能与通过某些途径拮抗Snail介导的End MT所参与的纤维增生性闭塞性血管损伤有关。Objective： To investigate the effects and mechanism of Chinese herbal compound in Bushen Yijing Therapy on endothelial to mesenchymal transition （EndMT） in systemic seleroderma. Methods： Mice model of scleroderma was induced by bleomycin ,at the same time the No. 2 receipt of Ruanpi Decoction（ Chinese herbal compound） in Bushen Yijing prescription was given to the modeling mice for intervention. After 4 weeks, the pathological changes of skin tissue in mice were observed by HE staining,and the expression level of α-SMA was detected by immunohistochemistry. EndMT model of vascular endothelial cells induced by TGF-β1was established in vitro. After treatment with medicated serum from Bushen Yijing Therapy for 48 hours,the expression of α-SMA in vascular endothelial cells was detected by immunofluorescence, the expression levels of CD31 ,FSPI and Snail-1 in vascular endothelial cells were determined by Western Blot. Results： The results of HE staining showed that in the skin fibrosis area of mice in the model group, the collagen fibers obviously increased and the space between collagen narrowed, and the capillary was obviously less than that in normal skin tissue. Compared with the model group, the skin collagen fibers of mice in Chinese herbal compound group were less,and the space was wider and the capillary was more. The results of immunohistochemical staining showed that the expression of α-SMA in vascular endothelial cells of mice skin tissue in the model group was higher than that in the normal control group, and the positive expression of ot-SMA in Chinese herbal compound group was obviously decreased. The results of immunofluorescence showed that EndMT model was established successfully via the TGF-α1 induction. The high expression of α-SMA in cell was observed in EndMT model group, and the endothelial ceils were transformed into the mesenchymal cells. After the treatment with medicated serum, the expression of α-SMA was significantly decreased. The results of Western Blot showed that the

关 键 词：系统性硬皮病 血管内皮细胞 补肾益精法 血管内皮间质转化 

分 类 号：R285.5[医药卫生—中药学]