朱砂七多糖体外抗猪传染性胃肠炎病毒作用研究  

In vitro Inhibition of Polygonum cillinerve Polysaccharide on Swine transmissible gastroenteritis virus(TGEV)

作　　者：胡蓉[1] 郭浩杰[1] 曹志[1] 盛洁[1] 郭抗抗[1] 宁蓬勃[1] 卿素珠[1] 张为民[1] 

机构地区：[1]西北农林科技大学动物医学院,杨凌712100

出　　处：《农业生物技术学报》2015年第2期253-261,共9页Journal of Agricultural Biotechnology

基　　金：陕西省科技攻关项目(No.2013K01-61-01)

摘　　要：猪传染性胃肠炎(swine transmissble gastroenteritis,TGE)是由猪传染性胃肠炎病毒(Swine transmissible gastroenteritis virus,TGEV)引起的一种高度传染性的猪病毒性疾病。为探讨朱砂七(Polygonum cillinerve)多糖对TGEV的体外抑制作用,本研究以猪(Sus scrofa)睾丸(swine testis,ST)细胞为实验材料,通过先加药后接毒、先接毒后加药及药物与病毒混合感作3种给药方式,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐法MTT法)和实时荧光定量PCR(q RT-PCR)检测不同浓度的朱砂七多糖对TGEV在ST细胞的体外抗吸附、直接杀灭和抑制增殖作用。MTT法结果显示,朱砂七多糖对ST细胞的最大无毒浓度(TC0)为20μg/m L;最大安全浓度(20μg/m L)时,对TGEV的抗吸附作用最好,抑制率达85.2%;对体外增殖的抑制率达78.4%;直接灭活作用抑制率达74.4%,且抑制率与浓度呈正相关。q RT-PCR检测结果表明,朱砂七多糖在3种给药方式下均能在转录水平显著降低TGEV N m RNA相对表达量,与病毒对照组相比差异极显著(P<0.01),并存在一定的剂量依赖性。研究结果表明,朱砂七多糖对TGEV在ST细胞上增殖有抑制作用,不同给药方式有所差异,其中以先加药后接毒给药方式的预防作用效果最佳。本研究初步证实了朱砂七多糖的体外抗TGEV作用,为朱砂七资源的开发和综合利用提供了依据。Swine transmissble gastroenteritis（TGE） is a kind of hyperinfected viral disease in pigs （Sus scrofa） induced by Swine transmissble gastroenteritis virus （TGEV）, and leads to death of large numbers of the pigs. The present study aimed to evaluate the effect of Polygonum cillinerve polysaccharide （PCP） on inhibition of TGEV. Based on the detection results of TCID50 of TGEV and maximum cytotoxic concentration （TCo） of PCP on in vitro growth of swine testicular （ST） cells, the PCP was pro- seeded, co- seeded or post- seeded with TGEV in ST cells, then the anti-virus adsorption, inactivation and inhibition of the proliferation of PCP were analyzed by 3- （4,5- Dimethylthiazol-2- yl）-2,5- diphenyltetrazolium bromide （MTT） method and quantitative Real-time PCR （qRT-PCR）. The detection of in vitro viral infection indicated that the value of TCIDso on TGEV was 10-6.25. After 72 h in vitro culture, ST cells were well-grown without the cytotoxic response when treated them with PCP at concentration of lower than 20 μg/mL compared with the blank controls, which determined the TCo of PCP was 20 ~tg/mL. According to the results of MTT, under the maximum safe concentration （20 pg/mL） of PCP, the anti-adsorption effect of PCP on TGEV in vitro was the best, the inhibition rate was 85.2%, the inhibition rate on in vitro virus proliferation was 78.47/o, while the inhibition rate of direct inactivation to TGEV was 74.4%, and the inhibition rate was positively correlated with the concentration. Moreover, qRT-PCR detection indicated that 3 administration methods of PCP could significantly reduce relative TGEV N mRNA expression at the transcriptional level （P〈0.01） compared with the virus control, and with a dose-dependent characteristic. The results suggested that all 3 administration methods of PCP could inhibit the proliferation of TGEV at ST cells in vitro, and the prevention effect of PCP was the best. This work demonstrated the in vitro anti-virus effect of PCP on TGE, and can provide

关 键 词：朱砂七多糖 猪传染性胃肠炎病毒(TGEV) 体外抑制 SWINE transmissible GASTROENTERITIS VIRUS (TGEV) 

分 类 号：S852.659.6[农业科学—基础兽医学]