川芎嗪对脂多糖诱导的人脐静脉内皮细胞高通透性的保护作用研究  

Protective effect of tetramethylpyrazine on lipopolysaccharide-induced high permeability of human umbilical vein endothelial cells

作　　者：张云霄 任婷 叶苗苗 王元元[1] 陶静[1] 李言[1] ZHANG Yun-xiao;REN Ting;YE Miao-miao;WANG Yuan-yuan;TAO Jing;LI Yan(Department of Pathophysiology,Department of Human Anatomy,Bengbu Medical College,Bengbu Anhui 233030,China;Institute of Clinic Anatomy,Department of Human Anatomy,Bengbu Medical College,Bengbu Anhui 233030,China)

机构地区：[1]蚌埠医学院病理生理学教研室,安徽蚌埠233030 [2]蚌埠医学院临床应用解剖研究所,人体解剖学教研室,安徽蚌埠233030

出　　处：《蚌埠医学院学报》2021年第2期141-145,共5页Journal of Bengbu Medical College

基　　金：国家自然科学基金面上项目(81673791);蚌埠医学院研究生创新课题(Byycxz1704);蚌埠医学院自然科学基金面上项目(BYKY1647,BYKY1656)。

摘　　要：目的:观察川芎嗪(tetramethylpyrazine,TMP)对脂多糖(lipopolysaccharide,LPS)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)通透性变化和细胞骨架相关蛋白表达影响。方法:以体外培养的HUVECs为实验对象,将细胞随机分6组:正常对照组,LPS组(1μg/mL LPS),TMP低、中、高剂量防治组(60μg/mL TMP+1μg/mL LPS、120μg/mL TMP+1μg/mL LPS、240μg/mL TMP+1μg/mL LPS),纯TMP组(240μg/mL TMP)。Transwell小室内建立内皮细胞通透性模型;细胞电压电阻仪测定各组细胞跨内皮细胞电阻值(TEER);FITC-葡聚糖法检测各组细胞通透性变化,Pull-Down实验拉下Rac1-GTPase活性蛋白,Western blotting检测各组细胞Rac1-GTPase、Rac1、LIMK1和p-LIMK1的蛋白表达。结果:3组不同浓度的TMP均可以抑制LPS诱导的内皮细胞TEER降低以及降低LPS导致的内皮细胞通透性的增加(P<0.05~P<0.01);Western blotting实验显示TMP可以有效抑制LPS诱导的HUVECs中的Rac1-GTPase、p-LIMK1、Rac1、LIMK1表达(P<0.05~P<0.01)。结论:TMP可以有效抑制LPS诱导的HUVECs通透性升高,其机制可能与其下调Rac1/LIMK1信号通路中Rac1-GTPase活性蛋白、LIMK1磷酸化蛋白表达有关。Objective:To investigate the effect of tetramethylpyrazine(TMP)on the permeability of human umbilical vein endothelial cells(HUVECs)induced by lipopolysaccharide(LPS)and the expression of cytoskeleton-related proteins.Methods:HUVECs cultured in vitro were randomly divide into normal control group,LPS group(1μg/mL LPS),TMP low,medium and high dose group(60μg/mL TMP+1μg/mL LPS,120μg/mL TMP+1μg/mL LPS,240μg/mL TMP+1μg/mL LPS),pure TMP group(240μg/mL TMP).Endothelial cell permeability model was established in Transwell chamber;cell voltage resistance meter was used to measure the trans-endothelial electrical resistance in each group;FITC-dextran method was used to detect the permeability change in each group,and Pull-Down experiment was used to collect Rac1-GTPase active protein,Western blotting was used to detect the protein expression of Rac1-GTPase,Rac1,LIMK1 and p-LIMK1 in each group.Results:TEER and permeability coefficient showed that three different concentrations of TMP inhibited the decrease of TEER induced by LPS in endothelial cells and decreased LPS-induced endothelial cell permeability(P<0.05 to P<0.01);Western blotting analysis showed that TMP could effectively inhibit the LPS-induced up-regulation of Rac1-GTPase,p-LIMK1,Rac1 and LIMK1 in HUVECs(P<0.05 to P<0.01).Conclusions:TMP can effectively inhibit the increase of permeability of HUVECs induced by LPS,which may be related to the down-regulation of Rac1-GTPase active protein and LIMK1 phosphorylation protein expression in Rac1/LIMK1 signaling pathway.

关 键 词：川芎嗪 脂多糖 内皮细胞通透性 Rac1/LIMK1信号通路 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学]