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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-1IIIB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-1IIIB与C8166细胞融合有较好的抑制作用,EC50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 study the inhibitory activity and mechanism of HIV-1 in vitro on SMS-E and SMS-F. Methods MTT colorimetry was used to determine the toxicity of extract of Pinus kwangsiensis pineum on C8166 cells. The inhibitory effect of the extract on HIV-1 was tested by the inhibition of syncytia, p24 inhibition and MT-4 cell protection. HIV-1 replication intermediates detection, fusion blocking experiments and time-sharing drug testing to determine the mechanism of action of the extract. Results The SMS-E and SMS-F extracts of Pinus kiangsui pine extract had a good inhibitory effect on HIV-1, and the therapeutic index (TI) of replication of HIV-1IIIB in C8166 cells were 436.46 and 558.61, respectively. HIV replication intermediate product test showed that SMS-F treatment group ssDNA and Late-RT copy number was significantly lower than the control group, suggesting that the role of virus replication in the early; fusion blocking experiments showed that SMS-F chronic H9 / HIV-1IIIB and C8166 cell fusion has a good inhibitory effect, EC50 value of 71.03μg / mL, indicating that SMS-F role in the virus entry phase; time-sharing drug experiments showed that the extract has no effect on HIV-1 has entered the cell again proved SMS-F acts on HIV into the cell stage. Conclusion SMS-F of Pine, Pinus koraiensis pine extract has a strong inhibitory activity against HIV-1, and its mechanism of action is to inhibit the entry of HIV-1 into the cell stage.