circ-RELL1对脓毒症内皮细胞糖萼影响及其机制的研究  

Effect and mechanism of circ-RELL1 on glycocalyx of endothelial cells in sepsis

作　　者：柴彦 方奕鹏 张会娟 郭喆 王仲[2] Chai Yan;Fang Yi-peng;Zhang Hui-juan;Guo Zhe;Wang Zhong(School of Clinical Medicine,Tsinghua University,Beijing 100084,China)

机构地区：[1]清华大学临床医学院,北京100084 [2]清华大学附属北京清华长庚医院,北京102218

出　　处：《中国急救医学》2020年第12期1182-1185,共4页Chinese Journal of Critical Care Medicine

基　　金：江苏康缘药业股份有限公司委托专项课题(12020C6004)。

摘　　要：目的探讨circ-RELL1对脓毒症内皮细胞模型糖萼的变化及其机制。方法构建脓毒症细胞模型,分为正常组(C组)、脂多糖(LPS)组和3-甲基腺嘌呤(3-MA)组;实时荧光定量核酸扩增(real-time quantitative PCR,qPCR)检测系统检测circ-RELL1表达水平,采用Western Blot检测各组细胞自噬标志蛋白(light chain3,LC3)Ⅰ/LC3Ⅱ比例和p62的表达水平,酶联免疫吸附实验(ELISA)法检测上清中糖萼成分变化,将circ-RELL1表达水平与糖萼成分做Pearson相关分析。结果qPCR结果显示,与正常组比较,LPS组circ-RELL1表达升高(P<0.05);与LPS组比较,3-MA组circ-RELL1表达升高(P<0.05)。Western Blot结果显示,与C组比较,LPS组LC3Ⅰ/LC3Ⅱ比值升高(P<0.05),p62表达降低(P<0.05);3-MA干预可使LC3Ⅰ/LC3Ⅱ比值及p62表达的变化程度降低(与LPS组比较,P<0.05)。ELISA结果显示,与C组比较,LPS组上清中糖萼成分明显增多(P<0.01);3-MA干预可使糖萼破坏减少(与LPS组比较,P<0.05)。相关分析结果显示,circ-RELL1表达水平与糖萼成分释放呈正相关,但相关性较弱。结论circ-RELL1可促进脓毒症内皮细胞糖萼分解,其机制可能与自噬调控有关。Objective To investigate the effect and the mechanism of circ-RELL1 on glycocalyx of endothelial cells in sepsis.Methods Human umbilical vein endothelial cells(HUVEC)were cultured in vitro and were divided into normal group,lipopolysaccharide(LPS)group,and 3-methyladenine(3-MA)group.The expression level of circ-RELL1 was detected by qPCR,LC3Ⅰ/LC3Ⅱand p62 was detected by Western Blot;the glycocalyx in supernatant was detected by ELISA;the correlation of circ-RELL1 with glycocalyx was analyzed by Pearson correlation analysis.Results qPCR results showed that compared with the normal group,the expression of circ-RELL1 was increased in the LPS group(P<0.05);compared with the LPS group,the expression of circ-RELL1 was increased in the 3-MA group(P<0.05);Western Blot results showed that compared with C group,LC3Ⅰ/LC3Ⅱratio was higher(P<0.05),and expression of p62 was decreased in LPS group(P<0.05);intervention of 3-MA can make the change of LC3Ⅰ/LC3Ⅱratio and p62 less(compared with LPS group,P<0.05).ELISA results showed that compared with group C,glycocalyx components in the supernatant were significantly increased in LPS group(P<0.01);intervention of 3-MA resulted in the reduction of glycocalyx damage(compared with LPS group,P<0.05).Correlation analysis results showed that the expression level of circ-RELL1 was positively correlated with the release of glycocalyx components,but the correlation was weak.Conclusion Circ-RELL1 is related to glycocalyx decomposition in the endothelial cells and its potential mechanism may be related to the regulation of autophagy.

关 键 词：脓毒症 自噬 circ-RELL1 糖萼 

分 类 号：R459.7[医药卫生—急诊医学]