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目的:研究番石榴叶总皂苷对HIV-1病毒进入靶细胞的影响,为寻找新的HIV-1进入抑制剂提供实验依据。方法:SA-1型大孔树脂纯化获得番石榴叶总皂苷(TSGL),观察其对HIV-1包膜蛋白介导的MT-2和CHO-WT细胞融合的作用,采用MTT法检测其细胞毒性,通过ELISA、N-PAGE等方法分析TSGL对HIV-1 gp41六螺旋束形成的影响。结果:TSGL能显著抑制HIV介导的细胞融合,IC50为(7.33±0.40)μg/mL,在此剂量下对MT-2和CHO-WT两种细胞均无生长抑制作用。ELISA实验表明,TSGL能显著抑制HIVgp41六螺旋束(6-HB)的形成,25μg/mL时抑制率达到95.93%,而N-PAGE结果进一步确证了药物对6-HB的抑制作用。结论:番石榴叶中总皂苷能抑制HIV进入靶细胞,其作用机制可能与阻止HIV-1包膜蛋白gp41亚基六螺旋束的形成有关。
Objective: To study the effect of guava leaf total saponins on HIV-1 virus entry into target cells and provide experimental evidence for finding novel HIV-1 entry inhibitors. Methods: The TSGL of SA-1 macroporous resin was purified to observe its effect on HIV-1 envelope protein-mediated fusion of MT-2 and CHO-WT cells. MTT assay was used to detect the cells Toxicity was analyzed by ELISA, N-PAGE and other methods TSGL HIV-1 gp41 six-helix bundle formation. RESULTS: TSGL significantly inhibited HIV-mediated cell fusion with an IC50 of (7.33 ± 0.40) μg / mL. No inhibition of growth was observed with both TS-2 and CHO-WT cells at this dose. The results of ELISA showed that TSGL could significantly inhibit the formation of 6xHg of HIVgp41, and the inhibitory rate reached 95.93% at 25μg / mL. The results of N-PAGE confirmed the inhibitory effect of TSGL on 6-HB. CONCLUSION: Total saponins in guava leaves can inhibit the entry of HIV into target cells. The mechanism may be related to the prevention of the formation of the hexahelical bundle of gp41 subunit of HIV-1.