思茅松松塔提取物体外抗HIV-1研究  

Anti-HIV-1 activities of pine cone extract from Pinus kesiya var. langbianensis in vitro

作　　者：崔雪青[1,2] 刘熙 陈欢[1] 彭芳 罗荣华[1] 杨柳萌[1] 刘光明 王睿睿[1,3] 郑永唐[1] 

机构地区：[1]中国科学院和云南省动物模型与人类疾病机理重点实验室,中国科学院昆明动物研究所,云南昆明650223 [2]大理大学,药学与化学学院,云南大理671000 [3]云南中医学院,中药学院,云南昆明650500

出　　处：《中草药》2016年第11期1914-1918,共5页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81260632);国家科技重大专项课题(2012ZX10001-006)

摘　　要：目的研究思茅松松塔提取物（SMS-E和SMS-F）体外对HIV-1的抑制活性和作用机制。方法采用MTT比色法检测思茅松松塔提取物对C8166细胞的毒性；合胞体抑制实验、p24抑制实验和MT-4细胞保护作用检测提取物对HIV-1的抑制作用。采用HIV-1复制中间产物检测、融合阻断实验和分时给药实验判断提取物的作用机制。结果思茅松松塔提取物SMS-E和SMS-F对HIV-1有很好的抑制作用，对HIV-1_IIIB在C8166细胞中复制的治疗指数（TI）分别为436.46和558.61。HIV复制中间产物检测实验显示SMS-F处理组ssDNA和Late-RT的拷贝数明显低于对照组，提示作用于病毒复制的早期；融合阻断实验显示SMS-F对慢性H9／HIV-1_IlIB与C8166细胞融合有较好的抑制作用，EC_50值为71.03μg∥mL，说明SMS-F作用于病毒进入阶段；分时给药实验显示提取物对已进入细胞的HIV-1没有抑制作用，再次证明了SMS-F作用于HIV进入细胞阶段。结论思茅松松塔提取物SMS-F对HIV-1有很强的抑制活性，其作用机制为抑制HIV-1进入细胞阶段。Objective To investigate the anti-HIV-I activity and mechanism of pine cone extracts from Pinus kesiya var. langbianensis in vitro. Methods The cytotoxicity of pine cone extract from P. kesiya var. langbianensis was examined by MTT assay; Syncytium formation inhibition assay, p24 assay, and the HIV-l-induced MT-4 cell death assay were used to evaluate the anti-HIV-1 activity of pine cone extract from P. kesiya vat. langbianensis. HIV DNA species detection assay, cell fusion assays, and time of addition assay were used to determine the mechanisms of the extracts. Results Pine cone extract from P. kesiya var. langbianensis named SMS-F showed the good inhibition on H1V-1 in C8166 cells with TI value above 436.46 and 558.61, respectively. HIV-1 DNA species detection showed that ssDNA and late-RT were lower in the SMS-F treated group than those in the control group. The result implied that the mechanism of SMS-F might target the early stage of HIV-1 replication. SMS-F blocked the cell to cell fusion with ECs0 value 71.03 μg/mL and could not inhibit the replication of HIV-1 after entering the cell, which confirmed SMS-F targeting the HIV entry stage. Conclusion Pine cone extract from P. kesiya var. langbianensis shows the good inhibition on HIV-1 targeting the viral entry stage.

关 键 词：松塔 思茅松 艾滋病 HIV 融合阻断 病毒进入 

分 类 号：R285.5[医药卫生—中药学]