目的 研究血府逐瘀汤对转化生长因子-β1(TGF-β1)诱导的大鼠肺微血管内皮细胞(pulmonary microvascular en-dothelial cells,PMVECs)内皮间质转化(endothelial-to-mes-enchymal transition,EndMT)的影响,并从 TGF-β1/Smad 信号通路进一步分析其作用机制.方法 采用TGF-β1(5 μg· L-1)建立EndMT细胞模型.实验分组为正常组,模型组(TGF-β1,5 μg·L-1),血府逐瘀汤低、中、高浓度组(3、10、30 mg·L-1 XFZYD+5 μg·L-1 TGF-β1),SB 431542 阳性药组(5 μmol·L-1 SB 431542+5 μg·L-1 TGF-β1).观察细胞形态,鬼笔环肽染色观察细胞骨架,免疫荧光法观察内皮细胞标志物-血小板内皮细胞黏附分子1(CD31)和间质细胞标志物α-平滑肌肌动蛋白(α-SMA)的表达.蛋白免疫印迹法检测血府逐瘀汤对CD31、α-SMA、Snail、p-Smad2、p-Smad1/5蛋白表达的影响.结果 TGF-β1刺激后,细胞形态由鹅卵石形向梭形转变.CD31、p-Smad1/5表达明显减少,α-SMA、p-Smad2、Snail表达则显著升高.血府逐瘀汤干预后,与模型组相比,细胞形态不同程度的接近于正常组,CD31 和 p-Smad1/5 表达呈上升趋势,α-SMA、p-Smad2、Snail表达呈下降趋势.结论 血府逐瘀汤可以减轻大鼠肺微血管内皮细胞发生内皮间质转化,其作用可能是通过影响TGF-β1/Smad信号通路实现的.
Aim To investigate the effect of Xuefu Zhuyu decoction on transforming growth factor-β1(TGF-β1)-induced endothelial-to-mesenchymal transi-tion(EndMT)of pulmonary microvascular endothelial cells(PMVEC),and further analyze the mechanism related to the TGF-β1/Smad signaling pathway.Meth-od To construct an EndMT cell model,PMVEC was treated with TGF-β1(5 μg·L-1).The experimental groups were divided into normal,model(5 μg·L-1 TGF-β1),Xuefu Zhuyu decoction(3,10,30 mg· L-1 XFZYD+5 μg·L-1 TGF-β1),and SB 431542 positive drug group(5 μmol·L-1 SB 431542+5 μg ·L-1 TGF-β1).The morphology of the cells was ob-served,and the cytoskeleton was detected using phal-loidin staining.The expression of endothelial markers platelet endothelial cell adhesion molecule-1(CD31)and the mesenchymal markers α-smooth muscle actin(α-SMA)were observed by immunofluorescence as-say.The expression levels of CD31,α-SMA,Snail,p-Smad2,and p-Smad1/5 protein were detected by Western blot.Result With TGF-β1 incubation,the morphology shifted from cobblestones to shuttle-shaped.The expression of CD31 and p-Smad1/5 was reduced significantly,whereas the expression of α-SMA,p-Smad2 and Snail was raised.In contrast to the model group,the morphology of the cells tended to ap-proach that of the normal group in varying degrees after the treatment with Xuefu Zhuyu decoction.CD31 and p-Smad1/5 showed an increasing trend,while α-SMA,p-Smad2 and Snail showed a decreasing trend.Con-clusion Xuefu Zhuyu decoction attenuates EndMT of pulmonary artery endothelial cells,which may be a-chieved by affecting the TGF-β1/Smad signaling path-way.