脂筏依赖性内吞途径对轮状病毒感染细胞表面NHE3蛋白的调控作用  

Effects of lipid rafts dependent endocytosis pathway on regulation of NHE3 trafficking mechanism on the surface of rotavirus infected cells

作　　者：王鹏 秦红霞[2] 田一博 李萌萌[4] 焦佳伟 孙艺文 王世广 WANG Peng;QIN Hong-xia;TIAN Yi-bo;LI Meng-meng;JIAO Jia-wei;SUN Yi-wen;WANG Shi-guang(College of Medicine,Zhengzhou University of Industrial Technology,Zhengzhou 451100,Henan,China;Department of Preventive Dentistry,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan,China;Inspection Department,Medical School,Zhengzhou University,Zhengzhou 450052,Henan,China;Basic Medical Department,School of Nursing and Health,Zhengzhou University,Zhengzhou 450001,Henan,China)

机构地区：[1]郑州工业应用技术学院医学院,郑州451100 [2]郑州大学第一附属医院口腔预防科,郑州450052 [3]郑州大学医学院检验系,郑州450052 [4]郑州大学护理与健康学院基础教研室,郑州450001

出　　处：《医学研究生学报》2021年第2期131-134,共4页Journal of Medical Postgraduates

基　　金：河南省教育厅高校科技创新人才项目(20HASTIT047),郑州大学青年教师基础研究基金(JC202020016)。

摘　　要：目的轮状病毒(RV)可引起儿童急性腹泻症状,但机制均尚不明确。研究表达NHE3的Caco-2细胞经轮状病毒感染后表面NHE3蛋白的改变及其机制。方法表达HA-NHE3的单层极化Caco-2细胞分别加入等体积无血清培养液、RV病毒液(1×104 TCID50/mL)、脂筏破坏剂MβCD(10 mmol/L)和MβCD+RV液,分别为对照组、RV组、脂筏破坏剂甲基-β-环糊精(MβCD)组和MβCD+RV组,分别采用生物素化法,BCECF-AM法和Western Blot法测定细胞表面NHE3蛋白量,Na+-H+交换活性和Cdc42蛋白水平。结果与对照组相比,RV组细胞NHE3的生物活性(0.107±0.016)显著降低(P<0.01),而MβCD+RV组(0.243±0.011)和MβCD处理组(0.255±0.012)差异无统计学意义(P=0.214)。与对照组细胞表面NHE3的表达水平(0.600±0.025)对比,RV组(0.274±0.028)显著降低(P<0.01),而MβCD组(0.609±0.015)和MβCD+RV组(0.577±0.014)差异无统计学意义(P=0.220)。Western Blot法检测发现,与对照组细胞Cdc42蛋白表达水平(0.213±0.009)比较,RV组(0.421±0.020)显著提高(P<0.01),而MβCD+RV组(0.196±0.007)差异无统计学意义(P=0.172)。结论RV对NHE3蛋白量及其生物活性的调控机制可能与脂筏依赖性内吞途径有关。Objective Rotavirus(RV)can cause acute diarrhea in children,but the mechanism is not very clear.To observe the regulation mechanism of rotavirus on the bioactivity and protein levels of NHE3 on Caco-2 cells.Methods Caco-2 cells expressing NHE3 were randomly divided into the control group,RV group,methyl-β-cyclodextrin(MβCD)group and MβCD+RV group.NHE3 surface amount,Na+-H+exchange activity and protein levels of Cdc42 were determined by biotinylation,BCECF-AM and Western blot,respectively.Results Na+-H+exchange activity and surface amount of NHE3 in the control group is 0.261±0.006 and 0.600±0.025,respectively.When the cells were treated with RV,Na+-H+exchange activity(0.107±0.016)and surface amount(0.274±0.028)were significantly decreased compared with those in the control group.The protein levels of Cdc42 were increased(0.421±0.020)compared with those in the control group(0.213±0.009).However,the expression of Cdc42 in MβCD+RV group(0.196±0.007)was not significantly changed compared with that in the control group(P=0.172).Conclusion Lipid rafts dependent endocytosis pathway may be involved in the regulation of RV on NHE3 protein levels and biological activity.

关 键 词：轮状病毒 钠-氢交换蛋白3 脂筏 内吞途径 

分 类 号：R511[医药卫生—内科学]