三没食子酰吡喃葡糖作用于gp41抑制HIV与靶细胞的融合

目的：检测来源于蛇菰科植物日本蛇菰中的单体化合物三没食子酰吡喃葡萄糖（TGGP）抑制HIV与靶细胞融合的活性，并探讨其作用靶点。方法：用萃取和柱层析方法分离提纯TGGP，用酶联免疫测定法、天然聚丙烯酰胺凝胶电泳、分子排阻高效液相色谱法检测其抑制HIV gp41六螺旋束结构形成的作用，用细胞-细胞融合方法检测TGGP抑制 HIV进入靶细胞的活性。结果：酶联免疫测定法表明TGGP能有效地抑制gp41六螺旋束结构的形成，其半数抑制浓度（IC₅₀）为（1.37±0.19）µg・ml^-1。进一步用生物物理方法直观地确证了 TGGP抑制gp41六螺旋束结构形成的作用。在生物学活性方面，50µg・ml^-1的TGGP能有效地抑制HIV包膜糖蛋白介导的细胞-细胞融合。结论：HIV进入靶细胞的过程由跨膜糖蛋白亚基gp41介导。三没食子酰吡喃葡萄糖（TGGP）能作用于gp41，抑制HIV与靶细胞的融合。该化合物可望作为阴道外用杀微生物剂，预防HIV的性传播。

Objective: To observe the inhibitory effect of 1,2,6-Tri-O-galloyl-β-D-glucopyranose (TGGP) from Balanophora japonica Makino on human immunodeficiency virus (HIV) entry into the host cells and explore the mechanisms. Methods: TGGP was purified from Balanophora japonica Makino by n-hexane and ethyl acetate extraction and column chromatography. The inhibitory activity of TGGP on HIV gp41 six-helix bundle formation was measured with ELISA, N-PAGE and SE-HPLC, and the inhibitory effect of TGGP on HIV envelope grlycoprotein-induced cell-cell fusion was detected using a non-infectious cell-based assay. Results: TGGP inhibited HIVgp41 six-helix bundle formation, with an IC₅₀ of 1.37±0.19 µg/ml as determined by ELISA, and this activity was further confirmed by N-PAGE and SE-HPLC. TGGP at 25 µg/ml significantly inhibited syncytium formation between the effector(CHO-WT) and the target (MT-2) cells. Conclusion: The HIV transmembrane subunitgp41 mediates the entry of HIV into the target cells. TGGP can inhibit HIV fusion and entry into the target cells by inhibiting the formation of gp41six-helix bundles, suggesting the potential of TGGP as a microbicide to prevent sexual transmission of HIV.