中草药提取液对流感病毒诱导细胞凋亡的影响

探讨金银花、黄芩和连翘的提取液对甲3型流感病毒诱导狗肾传代细胞(MDCK细胞)凋亡的影响。方法采用金银花、黄芩和连翘3种中草药的提取液进行抗病毒试验,以细胞病变、姬姆萨染色在光镜下观察细胞形态学变化,Annexin V和PI双染用流式细胞仪测定细胞凋亡率,评价药物对甲3型流感病毒诱导MDCK细胞凋亡的影响。结果在MDCK细胞上甲3型流感病毒(血凝滴度为1∶64)的TCID50为0.65.100μL-1;实验浓度:金银花为1∶256(1.95g.L-1),黄芩为1∶64(7.81g.L-1),连翘为1∶64(7.81g.L-1)。在光镜下观察甲3型流感病毒对照组的MDCK细胞,可见被感染的MDCK细胞出现了细胞凋亡特有的形态学变化。Annexin V和PI双染经流式细胞仪测定显示3种药物实验组的细胞凋亡率金银花(28.13±3.30)%、黄芩(13.47±3.04)%、连翘(19.17±6.55)%]均显著低于病毒对照组(54.10±4.58)%](P<0.001)。结论 7.81g.L-1的黄芩及7.81g.L-1的连翘均能抑制100TCID50的甲3型流感病毒诱导MDCK细胞凋亡。1.95g.L-1的金银花不能够有效抑制100TCID50的甲3型流感病毒诱导MDCK细胞凋亡。

Effects of Chinese Herbal Extracts on Cell Apoptosis Induced by Influenza Viruses

Objective To investigate the effects of lonicerae,scutellaria and forsythiae extracts on influenza virus A3-induced apoptosis in Madin-Darby canine kidney(MDCK) cells.Methods The effects of lonicerae,scutellaria and forsythiae on influenza virus A3-induced MDCK cell apoptosis were investigated to assess the anti-virus activities of Chinese herbal extracts.Cytopathy was determined by Giemsa staining under the light microscope and cell apoptosis was detected using flow cytometry by double-labeling with Annexin V and PI.Results The TCID50 of H3N2 influenza virus(hemagglutinin titer of 1∶64) was 0.65·100 μL-1 in MDCK cells.Experimental concentrations of lonicerae,scutellaria and forsythiae were 1∶256(1.95 g·L-1),1∶64(7.81 g·L-1) and 1∶64(7.81 g·L-1),respectivelty.The infected MDCK cells showed apoptosis-specific morphological changes under the light microscope.Flow cytometry showed that lonicerae,scutellaria and forsythiae significantly decreased the apoptotic rates compared with control virus-infected cells (28.13±3.30)%,(13.47±3.04)% and(19.17±6.55)% vs(54.10±4.58)%,respectively;P<0.001].Conclusion Scutellaria extract(7.81 g·L-1) and forsythiae extract(7.81 g·L-1) can inhibit influenza virus A3-induced MDCK cell apoptosis,but lonicerae extract(1.95 g·L-1) has no significant inhibitory effect on MDCK cell apoptosis.