基于机械敏感性阳离子通道Piezo1的黄芪丹参药物血清对低流体切应力诱导的人内皮细胞功能紊乱的影响  

Effects of Huangqi Danshen Medicated Serum on Endothelial Dysfunction Induced by Low Fluid Shear Stress Based on Mechanosensitive Channel Piezo1

作　　者：张蒙[1] 龚觉晓[2] 毛晨晗 高昕[2] 张丞波 王新东[1,2] ZHANG Meng;GONG Juexiao;MAO Chenhan;GAO Xin;ZHANG Chengbo;WANG Xindong(The Third School of Clinical Medicine,Nanjing University of Chinese Medicine,Nanjing 210028,China;Affiliated Hospital of Traditional Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210028,China)

机构地区：[1]南京中医药大学第三临床医学院,江苏南京210028 [2]南京中医药大学附属中西医结合医院,江苏南京210028

出　　处：《中国中医药信息杂志》2022年第2期68-73,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(81973766、81403386);江苏省研究生科研与实践创新计划项目(SJCX21-0801);江苏省中医局科技项目(YB201922);全国中医药创新骨干人才项目(2019年);江苏省名老中医专家董其美传承工作室建设项目(2019年)。

摘　　要：目的观察黄芪丹参药物血清调控机械敏感性阳离子通道Piezo1对低流体切应力(FSS)诱导的人脐静脉内皮细胞(HUVEC)功能紊乱的影响,探讨其可能的作用机制。方法将细胞分为空白组、模型组、Piezo1激活剂组和芪参血清组,采用FSS加载分析设备加载低FSS建立细胞损伤模型,Piezo1激活剂组和芪参血清组分别给予Piezo1激活剂Yoda-1和黄芪丹参药物血清干预,空白组和模型组加入空白血清培养。MTT法检测细胞活力,流式细胞仪检测细胞凋亡,硝酸还原酶法检测细胞灌流液一氧化氮(NO)含量,均相竞争法检测内皮素-1(ET-1)含量,ELISA检测白细胞介素-1β(IL-1β)和肿瘤坏死因子α(TNF-α)含量,RT-PCR检测细胞内Piezo1、内皮型一氧化氮合酶(eNOS)m RNA表达,Western blot检测细胞内Piezo1、eNOS、pro-IL-1β蛋白表达。结果与空白组比较,模型组细胞活力显著降低(P<0.05),凋亡率显著升高(P<0.01),细胞灌流液NO含量显著减少,ET-1、IL-1β和TNF-α含量显著增加(P<0.01),细胞Piezo1、eNOS mRNA和蛋白表达显著降低(P<0.01),pro-IL-1β蛋白表达显著升高(P<0.01);与模型组比较,芪参血清组细胞活力显著升高(P<0.01),Piezo1激活剂组和芪参血清组细胞凋亡率显著降低(P<0.01),细胞灌流液中NO含量显著增加(P<0.01),ET-1、IL-1β和TNF-α含量显著减少(P<0.01),细胞Piezo1、eNOS mRNA和蛋白表达显著升高(P<0.01),pro-IL-1β蛋白表达显著降低(P<0.05,P<0.01)。结论黄芪丹参药物血清对低FSS诱导的HUVEC炎症和功能紊乱有保护作用,其机制与调控机械敏感性阳离子通道Piezo1有关。Objective To observe the effects of Huangqi Danshen medicated serum on the injury model of human umbilical vein endothelial cells(HUVEC)induced by low fluid shear stress(FSS)by regulating mechanosensitive Piezo1 channel;To discuss its possible mechanism.Methods The cells were divided into blank group,model group,Piezo1 activator group and Qishen serum group.FSS loading analysis equipment was used to load low FSS to establish cell damage model.Piezo1 activator group and Qishen serum group were treated with Piezo1 activator Yoda-1 and Huangqi Danshen medicated serum respectively.Blank group and model group were cultured with blank serum.MTT was used to detect cell viability,flow cytometry was used to detect cell apoptosis.The content of nitric oxide(NO)in cell perfusate was detected by nitrate reductase method;the content of endothelin-1(ET-1)was detected by the homogeneous competition method;the contents of interleukin-1β(IL-1β)and tumor necrosis factorα(TNF-α)were detected by ELISA;RT-PCR was used to detect the mRNAs expression of Piezo1 and endothelial nitric oxide synthase(eNOS);Western blot was used to detect the proteins expression of Piezo1,eNOS,and pro-IL-1βin cells.Results Compared with the blank group,the cell viability of the model group significantly decreased(P<0.05),the apoptosis rate significantly increased(P<0.01),the content of NO in the cell perfusate decreased,and the contents of ET-1,IL-1βand TNF-αsignificantly increased(P<0.01),the mRNAs and proteins expression of Piezo1,eNOS were significantly reduced(P<0.01),and the protein expression of pro-IL-1βsignificantly increased(P<0.01).Compared with the model group,the cell viability of Qishen serum group significantly increased(P<0.01),the apoptosis rate of Piezo1 activator group and Qishen serum group were significantly decreased(P<0.01),the content of NO in cell perfusate significantly increased(P<0.01),and the contents of ET-1,IL-1βand TNF-αwere significantly reduced(P<0.01),the mRNAs and proteins expression of Piezo1,eNOS were signific

关 键 词：黄芪 丹参 机械敏感性阳离子通道Piezo1 流体切应力 人脐静脉内皮细胞 

分 类 号：R285.5[医药卫生—中药学]