血府逐瘀汤对肺动脉内皮细胞间质转化的影响及机制  

Effect Xuefu Zhuyu decoction on endothelial-to-mesenchymal transition of pulmonary artery endothelial cells and its mechanism

作　　者：曾作梅 王昕玥 田雷瑜 崔力丹 郭健 陈俞材 ZENG Zuo-mei;WANG Xin-yue;TIAN Lei-yu;CUI Li-dan;GUO Jian;CHEN Yu-cai(School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100050,China)

机构地区：[1]北京中医药大学中医学院,北京100050

出　　处：《中国药理学通报》2024年第1期155-161,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金青年项目(No 82003983);北京中医药大学“解码中医”揭榜挂帅项目(No 2022-JYB-JBZR-003);分子药理和药物评价教育部重点实验室开放课题(No P202201)。

摘　　要：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 endothelial cells, PMVECs)内皮间质转化(endothelial-to-mesenchymal 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(5μg·L^(-1))建立EndMT细胞模型。实验分组为正常组,模型组(TGF-β1,5μg·L^(-1)),血府逐瘀汤低、中、高浓度组(3、10、30 mg·L^(-1)XFZYD+5μg·L^(-1)TGF-β1),SB 431542阳性药组(5μmol·L^(-1)SB 431542+5μg·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 transition(EndMT)of pulmonary microvascular endothelial cells(PMVEC),and further analyze the mechanism related to the TGF-β1/Smad signaling pathway.Method To construct an EndMT cell model,PMVEC was treated with TGF-β1(5μg·L^(-1)).The experimental groups were divided into normal,model(5μg·L^(-1) TGF-β1),Xuefu Zhuyu decoction(3,10,30 mg·L^(-1) XFZYD+5μg·L^(-1) TGF-β1),and SB 431542 positive drug group(5μmol·L^(-1) SB 431542+5μg·L^(-1) TGF-β1).The morphology of the cells was observed,and the cytoskeleton was detected using phalloidin staining.The expression of endothelial markers platelet endothelial cell adhesion molecule-1(CD31)and the mesenchymal markersα-smooth muscle actin(α-SMA)were observed by immunofluorescence assay.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 ofα-SMA,p-Smad2 and Snail was raised.In contrast to the model group,the morphology of the cells tended to approach that of the normal group in varying degrees after the treatment with Xuefu Zhuyu decoction.CD31 and p-Smad1/5 showed an increasing trend,whileα-SMA,p-Smad2 and Snail showed a decreasing trend.Conclusion Xuefu Zhuyu decoction attenuates EndMT of pulmonary artery endothelial cells,which may be achieved by affecting the TGF-β1/Smad signaling pathway.

关 键 词：血府逐瘀汤 肺微血管内皮细胞 内皮间质转化 TGF-Β1 TGF-Β1/SMAD信号通路 肺动脉高压 

分 类 号：R-332[医药卫生] R285.5R322.13R322.35R544