摘要
目的 :检测小檗碱(Berberine)对流感病毒A/PR/8/34(H1N1)感染的脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVECs)细胞骨架及通透性的影响。方法 :常规鸡胚增殖传代甲型流感病毒A/PR/8/34,-80℃冰箱保存,空斑实验检测病毒滴度;将HUVEC接种于96孔板,MTT法检测小檗碱对HUVEC的细胞毒性;用30 MOI A/PR/8/34病毒株刺激病毒组和小檗碱给药组HUVECs,小檗碱组加入5μg/m L小檗碱溶液,分别于给药的第0、2、6、12、24、36小时检测各组电阻值;激光共聚焦检测观察各组细胞骨架形态。结果 :A/PR/8/34的病毒滴度为(2~3)×108 PFU/m L;小檗碱对HUVEC最大无毒浓度为10μg/m L,细胞存活率约为94.05%;小檗碱组(5μg/m L)较病毒组细胞通透性显著降低(P<0.05),且与正常组相差不大(P>0.05);小檗碱组与病毒组相比,HUVEC细胞内球状肌动蛋白聚合成丝状肌动蛋白(filamentous actin,F-actin)分布于细胞膜,应力纤维减少,且细胞边缘恢复平整,呈现出不规则多边形,与正常组细胞表现趋向一致。结论 :小檗碱具有改善流感病毒感染后的内皮细胞骨架重构、细胞收缩和细胞形态改变的作用,从而改善流感病毒感染后内皮细胞通透性的升高。
Objective ：To investigate the effect of berberine on the cytoskeleton and permeability of human umbilical vein endothelial cells（HUVECs）infected with influenza virus A/PR/8/34（H1 N1）.Methods ：Influenza virus A/PR/8/34 were passaged in embryonated chicken eggs,and samples of allantoic fluid were stored at-80 ℃.Virus titer was measured by plaque assay.HUVEC were seeded in 96-well plates.MTT assay was used to detect the toxicity of berberine to HUVEC.The HUVECs in the virus and berberine group of HUVEC were stimulated with 30 MOI A/PR/8/34,and the cells in the berberine group were then added with 5 μg/m L berberine solution.The resistance values were measured at 0,2,6,12,24 and 36 h,respectively.The cytoskeleton morphology was observed by laser confocal microscopy.Results ：The virus titer of influenza virus A/PR/8/34 is（2-3）×10^8 PFU/m L.The maximum non-toxic concentration of berberine to HUVEC was 10 μg/m L,and the cell survival rate was about 94.05%.The permeability of the berberine group was significantly lower than that of the virus group（P〈0.05）,and the difference was not significant with the non-infected group（P〉0.05）.Compared with the virus group,the filamentous actin（F-actin）in HUVECs of the berberine group was distributed in the cell membrane,with the stress fibers decreased,and the edge of the cells recovered to flat Irregular polygons,consistent with the performance of non-infected cells.Conclusion ：Berberine has the effect of improving skeleton remodeling,cell shrinkage and cell morphological changes of endothelial cells infected with influenza virus,which can protect the endothelial cells from permeability increase induced by influenza virus infection.
出处
《辽宁中医药大学学报》
CAS
2017年第12期28-31,共4页
Journal of Liaoning University of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81473520)
关键词
小檗碱
流感病毒
细胞骨架
细胞通透性
berberine
influenza virus
cytoskeleton
cell permeability
作者简介
陈晓鸣（1992-），女，河南封丘人，硕士研究生，研究方向：感染性疾病的中医药防治。;通讯作者：吴莹（1980-），女，四川江邮人，副教授，博士，研究方向：中西医结合防治病毒性疾病。
