VZV对Schwann细胞朊蛋白糖基化的影响及甲钴胺的调节作用  

Effect of VZV on the cellular prion protein glycosylation of Schwann cells and the regulation of methylcobalamin

作　　者：许纲[1] 周朝生[1] 张雨 唐维桢[1] 程超[1] 许洁[1] 徐刚[1] 王晓梅[1] Xu Gang;Zhou Chaosheng;Zhang Yu;Tang Weizhen;Cheng Chao;Xu Jie;Xu Gang;Wang Xiaomei(Zoster-Associated Pain Center,Affiliated Tenth People′s Hospital of Tongji University,Shanghai Tenth People′s Hospital,Shanghai 200072,China)

机构地区：[1]同济大学附属第十人民医院上海市第十人民医院带状疱疹神经痛诊疗中心,上海200072

出　　处：《中华实验和临床病毒学杂志》2021年第3期266-271,共6页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金(81771209)。

摘　　要：目的探究水痘-带状疱疹病毒(varicella-zoster virus,VZV)对人Schwann细胞(human Schwann cells,hSC)朊蛋白(cellular prion protein,PrPC)糖基化特征的影响,及甲钴胺(methylcobalamin,MeB12)的调节作用。方法以感染复数1.0的VZV感染细胞48 h,加入250μg/ml的MeB12培养48 h,用抗体3F4分别包被上清和沉淀中PrPC,凝集素-ELISA法筛查PrPC的糖基化特征,并测定上清中超氧化物歧化酶(superoxide dismutase,SOD)活性与丙二醛(malondialdehyde,MDA)含量。结果VZV感染组细胞上清与沉淀中的PrPC聚糖比例与未感染组比较有明显变化,总聚糖比分别为1∶1.5和1∶2.6(F=24.18,P<0.001,LSD-t=8.27,P<0.001),提示VZV感染后PrPC稳定性下降,相应的SOD活性(4.43±2.05 U/mg)与未感染组(14.23±1.27 U/mg)比较明显下降(F=18.19,P=0.001,LSD-t=6.54,P<0.001),MDA水平(11.17±1.89 nmol/mg)与未感染组(3.73±0.35 nmol/mg)比较明显升高(F=30.70,P<0.001,LSD-t=8.25,P<0.001),差异均有统计学意义。加入MeB12后,VZV感染细胞沉淀中的聚糖较VZV感染未加药组有明显增加,总聚糖比为1∶2.4,提示MeB12增强了PrPC的稳定性,相应地SOD活性明显增高(11.07±2.07 U/mg,LSD-t=4.42,P=0.002),MDA水平明显下降(5.23±0.96 nmol/mg,LSD-t=6.58,P<0.001),与感染未加MeB12组比较差异有统计学意义。结论VZV可改变hSC中PrPC的糖基化特征,而MeB12可调节PrPC的糖基化特征,增强PrPC的稳定性,从而提高hSC的抗氧化能力。Objective To explore the effects of varicella-zoster virus(VZV)on the glycosylation characteristics of cellular prion protein(PrPC)in human Schwann cells(hSC)and the regulation of methylcobalamin(MeB12).Methods The hSC were inoculated with VZV at 1.0 multiplicity of infection for 48 hours,then 250μg/ml of MeB12 were added and cultured for 48 hours.PrPC from the supernatant and sediment were coated with anti-PrPC antibody(3F4)respectively and subjected to screening for glycans by sandwich lectin-ELISA.Meanwhile,the superoxide dismutase(SOD)and malondialdehyde(MDA)from the supernatant were detected by diagnostic reagent kit.Results The ratio of PrPC glycans in the supernatant to sediment of VZV-infected cells was found to be significantly different compared with those in the VZV-non-infected cells.The overall glycans ratios of the supernatant to the sediment was 1∶2.6 in the uninfected cells,while the ratio was 1∶1.5 in the VZV-infected cells(F=24.18,P<0.001,LSD-t=8.27,P<0.001),suggesting that stability of PrPC decreased after VZV infection,and correspondingly the activity of SOD(4.43±2.05 U/mg)was significantly reduced in the VZV-infected hSCs compared with those(14.23±1.27 U/mg)in the uninfected cells(F=18.19,P=0.001,LSD-t=6.54,P<0.001),the level of MDA(11.17±1.89 nmol/mg)was significantly elevated in the VZV-infected hSCs compared with those(3.73±0.35nmol/mg)in the uninfected cells(F=30.70,P<0.001,LSD-t=8.25,P<0.001).When the VZV-infected cells were added with 250μg/ml MeB12,glycans in the sediment of infected cells significantly increased compared with those in the VZV-infected cells without MeB12,the overall glycans ratio of the supernatant to the sediment was 1∶2.4,suggesting that MeB12 improved the stability of PrPC.Moreover,SOD activity(11.07±2.07 U/mg)was significantly increased(LSD-t=4.42,P=0.002),MDA level(5.23±0.96 nmol/mg)was significantly decreased(LSD-t=6.58,p<0.001)in the VZV-infected cells added with MeB12 compared with those in the VZV-infected cells without MeB12.Conclusions The gly

关 键 词：水痘-带状疱疹病毒 SCHWANN细胞 细胞型朊蛋白 糖基化 甲钴胺 

分 类 号：R752.12[医药卫生—皮肤病学与性病学]