藏药甘青乌头生物碱抗单纯疱疹病毒Ⅱ型体内外作用  

Evaluation of antiviral activity of Tibetan medicine aconitum tanguticum alkaloid against herpes simplex virus typeⅡin vitro and in vivo

作　　者：张春江[1] 李薇[2] 孙振鹏[2] 赵星文[2] 

机构地区：[1]兰州大学生命科学学院,甘肃兰州730000 [2]兰州生物制品研究所,甘肃兰州730046

出　　处：《兰州大学学报（医学版）》2010年第4期1-6,共6页Journal of Lanzhou University（Medical Sciences）

基　　金：教育部春晖计划(Z2008-1-62005);中央高校基本科研业务费专项资金(1zujbky-2010-61)资助项目;甘肃省科技支撑计划(090NKCA131);甘肃省中医药管理局中医项目(GZK-2010-35)

摘　　要：目的探讨藏药甘青乌头脂溶性生物碱（ALA）和水溶性生物碱（AWA）抗单纯疱疹病毒Ⅱ型（HSV-2）的作用和机制。方法四甲基偶氮唑蓝法测定细胞毒性,细胞病变作用法和蚀斑法测定甘青乌头生物碱体外抗HSV-2活性,以半数有效浓度和治疗指数为评价指标;荧光定量聚合酶链式反应法（FQ-PCR）和蚀斑法考察ALA和AWA体外抗HSV-2作用机制。用HSV-2建立小鼠脑炎模型,对AWA体内抗HSV-2的活性进行评价。结果 ALA和AWA对Vero细胞的半数中毒浓度分别为3.57和7.87mg/mL。ALA和AWA半数有效质量浓度分别为4.99和2.81 mg/mL,治疗指数分别为0.72和2.80。ALA和AWA均能直接灭活HSV-2,能够显著抑制HSV-2的感染性。ALA对吸附到非洲绿猴肾细胞的HSV-2没有抑制作用,能微弱抑制HSV-2大分子的增殖,对HSV-2 DNA的复制没有作用。AWA能够抑制HSV-2吸附,抑制HSV-2大分子的增殖,抑制HSV-2 DNA的合成,抑制倍数达10~2。AWA对小鼠的半数中毒质量浓度为0.28mg/kg。AWA延长了小鼠的平均存活时间,对小鼠的死亡显示出10%的保护率。结论 AWA抗HSV-2的活性大于ALA,ALA主要通过直接灭活HSV-2,AWA通过抑制HSV-2复制循环的各个环节起作用。Objective To evaluate the antiviral effect and mechanisms of Tibetan herbal medicine Aconitum Tanguticum lipid-soluble alkaloid and water-soluble alkaloid against herpes simplex virusⅡ.Methods The cytotoxicity was assayed by methyl thiazolyl tetrazolium colorimetry, the antiviral effective rate was investigated by using cytopathic effect and plaque reduction assay. Median effective concentration and treatment index were used to estimate index of antiviral activity. Possible mode of action and molecular mechanism were investigated with fluorescence quan- titative polymerase chain reaction and plaque reduction assay.Mice encephalitis herpes models were established to evaluate the antiviral efficacy in vivo.Results The median cytotoxity concentration of lipid-soluble alkaloid and water-soluble alkaloid value was 3.57 and 7.87 mg/mL.The median effective concentration was 4.99 and 2.81 mg/ml by respectively using plaque reduction assay.The therapeutic index was 0.72 and 2.80 either.Aconitum tanguticum alkaloid diminished virus infectivity significantly,but lipid-soluble alkaloid had no effect on virus attachment to Vero cell and HSV-2 DNA replication.Lipid-soluble alkaloid weakly inhibited biosynthesis of HSV-2 in cells.Water-soluble alkaloid not only inhibited virus attachment and biosynthesis,but also inhibited DNA replication of HSV-2 and produce 102 times reduction in virus.The lethal dose 50 of water-soluble alkaloid was 0.28 mg/kg.Water-soluble alkaloid prolonged mean survival times and reduced 10%protection rate of mice.Conclusion The effect of antiviral of water-soluble alkaloid is stronger than that of lipid-soluble alkaloid,which can diminish virus infectivity directly.The antiviral mechanisms of water-soluble alkaloid are related to different stage of virus replication.

关 键 词：藏药 甘青乌头 生物碱 单纯疱疹病毒Ⅱ型 细胞病变作用法 蚀斑法 荧光定量聚合酶链式反应法 

分 类 号：R285.5[医药卫生—中药学]