三将丹体外抗HIV感染的活性研究及其机制探索

In vitro anti-HIV activity and its possible mechanism of an extract of herb medicines, SanJiangDan

目的研究三将丹体外抗HIV感染的活性及潜在机制。方法用蒸馏法从三将丹中抽提出主要活性成分，在细胞水平采用3种亚型的HIV假病毒（B亚型、C亚型、CRF01_AE亚型）对其体外抗病毒感染的活性进行评价。进一步研究三将丹对HIV宿主免疫细胞表面受体以及细胞因子分泌能力的影响，以探寻其可能的抗病毒机制。最后采用4种不同来源的细胞系（上皮细胞Caco-2细胞，TZM细胞，肝来源细胞Huh7细胞，淋巴细胞Jurkat—T细胞）对三将丹进行细胞水平的安全性评价。结果三将丹在1．6ms／ml和0．16mg／ml的浓度时能够很好地抑制HIV假病毒对细胞的感染，当浓度为0．16mg／ml时的抑制率为30．9％（B亚型）、36．6％（C亚型）以及65．0％（CRF01_AE亚型）。而当浓度为1．6mg／ml时，抑制率上升为96．4％（B亚型）、97．4％（C亚型）以及99．5％（CRF01_AE亚型），且不对宿主细胞造成毒性，同时三将丹能够抑制TZM-bl细胞表面CIM、CXCR4以及CCR5的表达，且三将丹能够诱导T细胞分泌IL-2。结论三将丹在体外能够在不对宿主细胞造成毒性的情况下，抑制HIV假病毒的感染，同时还能够促进诱导T细胞分泌IL-2，对免疫细胞具有一定的调理作用，这可能与三将丹抗HIV的活性机制有关。

Objective To study in vitro anti-HIV activity of an extract of herb medicines, San- JiangDan, and the possible mechanism. Methods The main active ingredient of SanJiangDan was extracted by distillation. Three subtypes of HIV pseudovirus （B subtype, C subtype, CRF01_AE subtype） were used to evaluate the anti-HIV activity of SanJiangDan extract in vitro. The possible mechanism was evaluated through analyzing the effects of SanJiangDan extract on the expression of surface receptors and cytokines by T cells. The cytotoxicity of SanJiangDan extract was detected by using four different sources of cell lines inclu- ding epithelial cells Caco-2 cells, TZM cells, Huh7 cells derived from liver cells and lymphocyte Jurkat-T cells. Results SanJiangDan extract effectively inhibited the infection of HIV pseudoviruses at concentra- tions of 1.6 mg/ml and 0. 16 mg/ml. The inhibition rates were 30.9% , 36.6% and 65.0% for B subtype, C subtype and CRF01_AE subtype respectively at the concentration of 0.16 mg/ml. As the concentration in- creased to 1.6 mg/ml, the inhibition rates increased to 96.4% （B subtype）, 97.4% （C subtype） and 99.5% （CRF01_AE subtypes）, but no toxicity to host cells was detected. Moreover, SanJiangDan extract inhibited the expression of HIV surface receptors including CIM, CXCR4 and CCR5 on TZM-bl cells, but enhanced IL-2 production. Conclusion SanJiangDan extract could inhibit HIV pseudovirus infection with- out causing cytotoxicity to host cells in vitro. The possible mechanism might be associated with the reduced expression of CIM, CXCR4 and CCP,5 and enhanced secretion of 1L-2 as well.