黄连提取物对TGEV的体外抑制效果

Effect of Coptis Rhizoma Extract on Inhibition of TGEV in vitro

旨在明确黄连提取物(CRE)体外抑制TGEV作用效果。CRE通过药物和病毒作用后再添加、先感染病毒再加药物、先添加药物再感染病毒3种方式作用于PK-15细胞,分别标记为试验1、2、3组,作用结束后采用MTT检测细胞活率,RT-PCR检测TGEV N基因和抗病毒相关因子(IFN-α、IFN-β、IFN-γ、IRF-3)的表达来评价CRE抑制TGEV作用效果。结果显示:1×10^-4 g/L的CRE通过3种方式处理的PK-15细胞其活率分别为54.12%、40.69%、87.67%,均极显著高于病毒组(P<0.01),且试验3组极显著高于试验1组和试验2组(P<0.01);TGEV N基因转录倍数分别降为病毒组的0.438、0.603、0.264倍,极显著低于病毒组(P<0.01);IFN-α、IFN-β、IRF-3转录量极显著高于空白组、病毒组和药物对照组(P<0.01),IFN-γ显著(P<0.05)或者极显著高于空白组和病毒组(P<0.01)。从而表明CRE通过3种方式可以有效保护宿主细胞,抑制TGEV增殖,激发抗病毒相关因子IFN-α、IFN-β、IFN-γ和IRF-3的表达来参与抗病毒作用,且先添加药物再感染病毒的作用方式效果最优。

For exploring the effect of Coptidis Rhizoma extract(CRE)on inhibition of TGEV in vitro.The effect of CRE on PK-15 was systematically studied by three methods including adding after the medicine and virus action,infected with the virus before adding the medicine,adding the medicine before infected with the virus,and it was separately marked as test group 1,2 and 3.After inculation,the cellular activity was tested by MTT assay,expression of TGEV N gene and antiviral related factors(IFN-α,IFN-β,IFN-γand IRF-3)were tested by RT-PCR assay to evaluate the effect of CRE on inhibition of TGEV.The results showed that the cellular activity of PK-15 cells treated with CRE at 1×10^-4 g/L concentration by the three methods was 54.12%,40.69%and 87.67%,respectively,which were significantly higher than that in the virus group(P<0.01).And the test group 3 was significantly higher than the test group 1 and the test group 2(P<0.01).The transcription ratio of GEV N gene was reduced to 0.438,0.603,and 0.264 times in the virus group,which was significantly lower than that in the virus group(P<0.01).The transcription ratio of IFN-α,IFN-βand IRF-3 were significantly higher than those in the blank group,the virus group and the drug control group(P<0.01),IFN-γwas also significantly(P<0.05)or extremely significantly higher than the blank group and virus group(P<0.01).In conclusion,the CRE by the three methods may effectively protect host cells,inhibit TGEV proliferation,stimulate the expression of antiviral related factors IFN-α,IFN-β,IFN-γand IRF-3 participating in antiviral effect.Finnally,adding the medicine before infecting virus was an optimal method.