复方参术对TGEV感染仔猪小肠黏膜上皮细胞凋亡因子Bcl-2/Bax的影响  

Effects of compound shenzhu on the production of Bcl-2/Bax induced by TGEV in swine intestinal epithelial cells

作　　者：王雪飞 王炼[1,2] 李存法[1] 刘永录[1] 杨英[2] WANG Xuefei;WANG Lian;LI Cunfa;LIU Yonglu;YANG Ying(Henan University of Animal Husbandry and Economy,Zhengzhou 450011;College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohehot,010018)

机构地区：[1]河南牧业经济学院,河南郑州450011 [2]内蒙古农业大学兽医学院,内蒙古呼和浩特010018

出　　处：《现代牧业》2018年第1期5-9,共5页Modern Animal Husbandry

基　　金：内蒙古自治区2014年博士研究生科研创新重点项目(B20141012908Z);河南省科技攻关项目(162102110033)

摘　　要：探讨中药复方参术对感染猪传染性胃肠炎病毒(TGEV)的猪小肠粘膜上皮细胞(SIEC)中凋亡因子Bcl-2/Bax表达水平的影响。体外增殖TGEV,用TGEV感染SIEC为模型进行体外试验,试验分为复方参术组、空白对照组、模型组、苦参碱组,分别于加药后2h、4h、8h、12h、24h、48h、72h,用荧光显微镜观察细胞凋亡情况,应用qRT-PCR和western blot检测Bcl-2与Bax表达水平。与空白对照组相比,模型组的SIEC发生了明显的细胞病变效应(CPE);与模型组比,复方参术组与苦参碱组细胞病变得到改善,复方参术组效果更加明显;与空白对照组相比,模型组Bcl-2 mRNA表达量极显著减少(P<0.01),复方参术组表达量增加极显著(P<0.01);与空白对照组相比,模型组Bax mRNA表达量极显著上调(P<0.01),复方参术组表达量减少(P<0.01)。这与Bcl-2/Bax蛋白表达水平检测到的结果一致。复方参术很可能通过上调SIEC Bcl-2 mRNA和蛋白的表达,下调Bax mRNA和蛋白的表达,从而抑制TGEV引起的SIEC凋亡。The objective of this study was to investigate the effects of compound shenzhu on the expression of Bcl-2/Bax mRNA and protein in swine intestinal epithelial cell(SIEC)infected with transmissible gastroenteritis virus(TGEV).TGEV proliferates in PK-15 and then make this virus infected SIEC in vitro.Test was divided into compound shenzhu group,blank control group,model group and Matrine group.At 2h、4h、8h、12h、24h、48h and 72 hours post inoculation,the presence of apoptosis was observed by fluorescence microscopy.Meanwhile Bcl-2/Bax mRNA and protein expression levels were assessed by qRT-PCR and Western blot respectively.Compared with blank control group,SIEC in model group had obvious cytopathic effects(CPE).CPE in compound shenzhu group and Matrine group was less serious.Compound shenzhu group had better improvement effects than Matrine group.The expression of Bcl-2 mRNA decreased significantly in model group and increased significantly in compound shenzhu group compared with blank control group(P<0.01).However,compared with blank control group,Bax expression was markedly increased in model group and decreased in compound shenzhu group(P<0.01).Bcl-2/Bax protein expression results were consistent with gene expression results.Compound shenzhu might inhibit apoptosis in SIEC infected by TGEV through up-regulate transcription of Bcl-2 and down-regulate transcription of Bax.

关 键 词：复方参术 传染性胃肠炎病毒 猪小肠黏膜上皮细胞 Bcl-2 Bax 

分 类 号：S855[农业科学—临床兽医学]