秋水仙碱对LPS诱导内皮间质转化的影响及机制  

Effect of colchicine on LPS induced endothelial mesenchymal transformation and its mechanism

作　　者：郭俊 唐光能[2] 赵祺 曹政[2] 涂强 GUO Jun;TANG Guang-neng;ZHAO Qi;CAO Zheng;TU Qiang(Fuwai Hospital,Chinese Academy of Medical Sciences,Beijing 100037,China;Shiyan Taihe Hospital,Shiyan Hubei 442000,China)

机构地区：[1]中国医学科学院阜外医院,北京100037 [2]十堰市太和医院,湖北十堰442000

出　　处：《中国药理学通报》2024年第2期243-248,共6页Chinese Pharmacological Bulletin

基　　金：湖北省创新群体项目(No 2022CFA036);湖北省教育厅指导性项目(No B2019111)。

摘　　要：目的研究秋水仙碱(colchicine,Col)对脂多糖(lipopolysaccharide,LPS)诱导的人脐静脉血管内皮细胞(human umbilical vein vascular endothelial cells,HUVECs)内皮间质转化的影响及相关机制。方法采用LPS处理HUVECs建立内皮间质转化(endothelial to mesenchymal transition,EndMT)模型。CCK-8法检测细胞增殖率,LDH实验检测细胞毒性,筛选最适药物浓度。将细胞分为正常对照组,正常对照+秋水仙碱(10 nmol·L^(-1))组,LPS(10 mg·L^(-1))模型组,LPS+秋水仙碱(10 nmol·L^(-1))组。倒置显微镜观察细胞形态学变化情况,Transwell实验检测细胞迁移能力,小管形成实验检测小管形成能力,Western blot检测内皮标志物(CD31/VE-cadherin)及间质转化标志物(α-SMA/FSP-1)表达情况;使用NF-κB信号通路抑制剂处理,检测相关信号通路变化情况。结果CCK-8及LDH实验显示,10 nmol·L^(-1)的Col为最适药物浓度;LPS可诱导细胞形态发生变化,Col在一定程度可逆转HUVECs的形态学变化;Transwell实验显示,LPS处理组HUVECs迁移能力明显增强(P<0.05),而Col可明显逆转这种现象(P<0.05);小管形成实验显示,LPS处理可抑制HUVECs小管形成能力(P<0.05),Col可改善LPS诱导的小管形成能力受损(P<0.05);Western blot结果显示,Col与LPS共同孵育后,CD31及VE-cadherin表达水平相比于模型组明显增加(P<0.05),α-SMA及FSP-1表达水平相比于模型组明显下降(P<0.05);在LPS诱导内皮细胞EndMT过程中,Col可抑制NF-κB/Snail信号通路激活。结论Col能有效抑制LPS诱导的EndMT,其机制与调控NF-κB/Snail信号通路有关。Aim To investigate the effect of colchicine on lipopolysaccharide(LPS)induced endothelial to mesenchymal transition(EndMT)in human umbilical vein vascular endothelial cells(HUVECs)and its related mechanisms.Methods The EndMT model was established by treating HUVECs with LPS.Cell proliferation rate was detected by CCK-8 assay,cytotoxicity was detected by LDH assay,and the optimal drug concentration was screened.The cells were divided into the normal control group,the normal control+colchicine(10 nmol·L^(-1))group,the LPS(10 mg·L^(-1))model group,and the LPS+colchicine(10 nmol·L^(-1))group.The morphologic changes of the cells were observed under an inverted microscope,the cell migration ability was detected by Transwell assay,and the ability of tube formation was analyzed by tube formation assay.The expression of endothelial markers(CD31/VE-cadherin)and mesenchymal cell markers(α-SMA/FSP-1)were detected by Western blot.NF-κB inhibitor was used to detect the changes in related signaling pathways.Results CCK-8 and LDH experiments showed that 10 nmol·L^(-1)colchicine was the optimal concentration.LPS could induce morphological changes in HUVECs,and colchicine could reverse morphological changes in HUVECs to a certain extent.Transwell experiment showed that the migration ability of HUVECs in the LPS treatment group was significantly enhanced(P<0.05),and colchicine could significantly reverse this phenomenon(P<0.05).Tube formation experiment showed that LPS decreased the endothelial tube formation ability of HUVECs(P<0.05),while colchicine treatment markedly improved LPS-induced tube formation defects(P<0.05).Western blot assay showed that after colchicine co-cultured with LPS,the expression levels of CD31 and VE-cadherin significantly increased compared with the model group(P<0.05),while the expression levels ofα-SMA and FSP-1 significantly decreased compared with the model group(P<0.05).During the induction of EndMT by LPS,colchicine could inhibit the activation of the NF-κB/Snail signaling pathway.Conclusions

关 键 词：脂多糖 内皮细胞 内皮间质转化 秋水仙碱 炎症 NF-ΚB信号通路 

分 类 号：R322.123[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学] R343.9R364.5R971.1