温阳化浊通络方体外抑制HIF-1a/Foxm1/smad3信号通路改善系统性硬化病肺微血管损伤  

Research on mechanism of Wenyang Huazhuo Tongluo formula inhibiting HIF-1a/Foxm1/smad3 pathway to improve pulmonary microvascular injury of systemic sclerosis

作　　者：卞博 苗青[2] 吴范武 范艺龄 孔金莉 卞华 李凯 BIAN Bo;MIAO Qing;WU Fan-wu;FAN Yi-ling;KONG Jin-li;BIAN Hua;LI Kai(Traditional Chinese Medical College,North China University of Science and Technology,Tangshan,Hebei 063210,China;Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Beijing University of Chinese Medicine,Beijing 100029,China;Zhang Zhongjing College of Chinese Medicine,Nanyang Institute of Technology,Nanyang,Henan 473004,China)

机构地区：[1]华北理工大学中医学院,河北唐山063210 [2]中国中医科学院西苑医院,北京100091 [3]北京中医药大学,北京100029 [4]南阳理工学院张仲景国医国药学院,河南南阳473004

出　　处：《中国药理学通报》2024年第11期2119-2123,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No 82074415);中原科技创新领军人才项目(No 234200510006)。

摘　　要：目的利用系统性硬化病(systemic sclerosis,SSc)肺微血管内皮细胞缺氧模型,研究温阳化浊通络方抑制内皮细胞的内皮-间充质转化(EndoMT)改善肺微血管损伤的分子机制。方法利用SSc患者血清培养正常人肺微血管内皮细胞构建SSc肺微血管内皮细胞,液体石蜡封闭法建立SSc肺微血管内皮细胞缺氧模型,并分别用温阳化浊通络方含药血清或HIF-1a抑制剂KC7F2处理,Western blot检测VE-cadherin、CD31、vimentin、HIF-1α、Foxm1、smad3、Tie-1以及vWF的蛋白表达水平,ELISA检测细胞培养液中的E-selectin和ICAM-1的浓度,双荧光素酶报告基因系统检测Foxm1基因启动子活性。结果与对照组相比较,模型组细胞中的VE-cadherin、CD31、HIF-1a、Foxm1、smad3、Tie-1和vWF蛋白表达水平显著降低,vimentin蛋白表达水平以及细胞培养液中E-selectin和ICAM-1的浓度显著增高,同时细胞形态呈现出明显的“纺锤体样”成肌纤维细胞形态,而温阳化浊通络方和KC7F2能够逆转这些缺氧诱导的蛋白表达水平,以及细胞形态的改变。结论温阳化浊通络方能够改善SSc肺微血管损伤,其作用机制可能是抑制HIF-1a/Foxm1/smad3通路介导的肺微血管内皮细胞EndoMT。Aim To investigate the molecular mechanisms of the Wenyang Huazhuo Tongluo formula in inhibiting endothelial-to-mesenchymal transition(EndoMT)of pulmonary microvascular endothelial cells and improving pulmonary microvascular injury in systemic sclerosis(SSc).Methods Pulmonary microvascular endothelial cells were cultured with serum from SSc patients to establish SSc pulmonary microvascular endothelial cells.A hypoxia model was established in SSc pulmonary microvascular endothelial cells using liquid paraffin sealing,and the cells were treated with the Wenyang Huazhuo Tongluo formula or HIF-1a inhibitor KC7F2.Western blot was used to detect the protein expression levels of VE-cadherin,CD31,vimentin,HIF-1α,Foxm1,smad3,Tie-1,and vWF.ELISA was used to measure the concentrations of E-selectin and ICAM-1 in cell culture medium.The luciferase reporter gene system was used to detect the promoter activity of the Foxm1 gene.Results Compared to the control group,the levels of VE-cadherin,CD31,HIF-1α,Foxm1,smad3,Tie-1,and vWF significantly decreased under hypoxic condition,while the levels of vimentin,E-selectin,and ICAM-1 significantly increased.In addition,the cell morphology exhibited a distinct“spindle-like”myoblast morphology.Treatment with the Wenyang Huazhuo Tongluo formula or KC7F2 reversed these changes in protein expression levels and cell morphology induced by hypoxia.Conclusion The Wenyang Huazhuo Tongluo formula improves pulmonary microvascular injury in SSc by inhibiting the HIF-1a/Foxm1/smad3 pathway-mediated EndoMT of pulmonary microvascular endothelial cells.

关 键 词：温阳化浊通络方 内皮-间充质转化 系统性硬化病 缺氧 肺微血管损伤 HIF-1a/Foxm1/smad3通路 

分 类 号：R285[医药卫生—中药学]