轮状病毒对NHE3蛋白水平及其生物活性的影响  

Effects of rotavirus on expression levels and bioactivity of NHE3

作　　者：王昕[1] 范媛媛[1] 王鹏[1] 李玲 潘雪[1] 范程程[3] 孟雨姗 蒋莉莉[1] WANG Xin;FAN Yuan - yuan;WANG Peng;LI Ling;PAN Xue;FAN Cheng - cheng;MENG Yu -shan;JIANG Li - li(Basic Department, Nursing School of Zheng- zhou University, Zhengzhou 450052, Henan, Chin)

机构地区：[1]郑州大学护理学院基础教研室,河南郑州450052 [2]郑州市第九人民医院姑息治疗暨安宁疗护病区,河南郑州450053 [3]郑州大学药学院,河南郑州450052

出　　处：《广东医学》2018年第7期971-973,共3页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81500433);河南省高等学校重点科研项目(编号:16A310003)

摘　　要：目的研究轮状病毒(RV)感染对表达NHE3的Caco-2细胞表面NHE3蛋白水平及其生物活性的影响。方法构建表达NHE3的Caco-2细胞模型,构建好的表达HA-NHE3的单层极化Caco-2细胞分为CT组和RV组,分别加入等体积无血清培养液和RV病毒液(MOI10),在37℃、5% CO_2条件下孵育60min,进行检测。BCECF-AM法和细胞表面NHE3生物素化法测定Na^+-H^+交换活性和细胞表面NHE3蛋白水平。结果 Na^+-H^+交换活性检测结果显示,与CT组(0.412±0.026)相比,RV感染可引起细胞NHE3的生物活性明显降低(0.147±0.011)(P<0.01)。细胞表面NHE3生物素化法检测发现,CT组和RV组间细胞NHE3蛋白总量没有发生明显变化(0.727±0.037 vs 0.732±0.020),差异无统计学意义(P>0.05)。然而,与CT组(0.374±0.015)相比,RV感染表达HA-NHE3的单层极化Caco-2细胞60 min后,细胞表面NHE3的表达明显降低(0.204±0.009),差异有统计学意义(P<0.01)。结论 RV感染性腹泻可能与调控肠道细胞表面NHE3的蛋白水平及其生物活性有关。Objective To investigate the effects of rotavirus on expression levels and bioactivity of NHE3 on Caco - 2 cells expressing NHE3. Methods Cell model of Caco - 2 cells expressing NHE3 was constructed. The single - lay- er polarized Caco - 2 cells expressing HA - NHE3were divided to the control group and RV group. Serum - free culture and RV were added to the control group and the RV group, respectively. Both groups were incubated at 37℃, 5% CO2. NHE3 activity and surface amount were determined by BCECF -AM and biotinylation, respectively. Results NHE3 ac- tivity and surface amount detected by Na ＋ - H ＋ exchanger 3 significantly decreased when the cells were treated with RV （0. 147 ±0. 011 ） compared with that of the control group （0. 412 ± 0. 026）. With regard to the total amount of NHE3, there was no difference between the control group and the RV group （0. 727 ± 0. 037 vs 0. 732 ± 0. 02, P 〉 0. 05 ）. After incubation, NHE3 expression in the RV group （0. 374 ±0. 015 ） was significantly lower than that in the control group （0. 204 ± 0. 009, P 〈 0. 01 ）. Conclusion Surface level and bioactivity of NHE3 may be associated for rotavirus diarrhea.

关 键 词：轮状病毒 钠-氢交换蛋白3 腹泻 

分 类 号：R373[医药卫生—病原生物学]