复方参术汤通过线粒体通路抑制TGEV诱导猪小肠黏膜上皮细胞凋亡的研究

Mitochondrial pathway-mediated inhibition effects of compound shenzhu on the TGEV-induced apoptosis of swine intestinal epithelial cells

为探讨中药复方参术汤干预猪传染性胃肠炎病毒(transmissible gastroenteritis virus,TGEV)诱导猪小肠黏膜上皮细胞(SIEC)凋亡的机制。本试验用TGEV感染SIEC进行体外试验。试验分为复方参术汤组、对照组、TGEV组、苦参碱组,分别于加药后2,4,8,12,24,48,72h收集样本,应用qRT-PCR检测Bcl-2与Bax mRNA转录水平,Western blot检测Bcl-2、Bax、Bid、Cyt-c、Caspase-9和Caspase-3的蛋白表达水平。结果显示,与TGEV组相比,复方参术汤下调TGEV感染SIEC的Bax、Caspase-9、Caspase-3、Cyt c及Bid蛋白表达;复方参术汤极显著上调Bcl-2mRNA的转录(P<0.01),极显著下调Bax mRNA的转录(P<0.01)。结果表明,复方参术汤能够抑制引起细胞凋亡的线粒体通路,从而预防TGEV引起的SIEC凋亡。

The objective of this study is to investigate the intervention mechanism of compound Shenzhu for the apoptosis of swine intestinal epithelial cells （SIEC） induced by transmissible gastroenteritis virus （TGEV）. Using TGEV to infect SIECwas as a model for in vitro test with compound Shenzhu group, blank control group, TGEV group and matrine group. Then using qRT-PCR method and Western blot method detected the expression level of Bcl-2, Bax mRNA and the expression level of Bcl-2, Bax, Bid, Cyt-c, Caspase-9 and Caspase-3 protein after 2,4,8,12,24,48 and 72 hours after drug treatments separately. Compared with TGEV group, the expression level of Bax,Caspase-9,Caspase-3,Cyt c and Bid protein in TGEV-infected SIEC was decreased by com- pound Shenzhu. Compound Shenzhu markedly increased the expression level of Bcl-2 mRNA （P〈 0.01） and significantly decreased the expression levels of Bax mRNA （P〈0.01）; The results indi- cated that compound Shenzhu could suppress the apoptosis of TGEV-infected SIEC via inhibiting mitochondrial pathway.