Faculty of Dentistry, Thammasat University, Pathum Thani 12120, Thailand.
Faculty of Medicine, Princess of Naradhiwas University, Narathiwat 96000, Thailand.
Molecules. 2022 Nov 16;27(22):7941. doi: 10.3390/molecules27227941.
Ellagic acid (EA) has a wide range of biological effects. The purpose of this study was to investigate the in vitro effects of EA on HIV-1 replication, viral enzyme activity and cytokine secretion by infected cells.
The anti-HIV-1 activity of EA in solution was determined in vitro using the infection of TZM-bl cells by the nano luciferase-secreting R5-tropic JRCSF strain of HIV-1, which allows for the quantification of viral growth by measuring nano luciferase in the culture supernatants. The effect of EA on the cytokine secretion of TZM-bl cells was determined by a multiplexed bead array after 48 h of HIV-1 exposure. The antiviral effect of EA in the gel formulation (Ellagel), as would be used for vaginal application, was investigated by the inhibition of infection of UC87.CD4.CCR5 cells with R5-tropic pBaLEnv-recombinant HIV-1.
EA in solutions of up to 100 µM was not toxic to TZM-bl cells. EA added either 1 h before or 4 h after HIV-1 exposure suppressed the replication of R5-tropic HIV-1 in TZM-bl cells in a dose-dependent manner, with up to 69% inhibition at 50 µM. EA-containing solutions also exhibited a dose-dependent inhibitory effect on HIV-1 replication in U87 cells. When EA was formulated as a gel, Ellagel containing 25 µM and 50 µM EA inhibited HIV-1 replication in U87 cells by 56% and 84%, respectively. In assays of specific HIV-1 enzyme activity, Ellagel inhibited HIV-1 integrase but not protease. EA did not significantly modulate cytokine secretion.
We conclude that EA either in solution or in a gel form inhibits HIV infection without adverse effects on target cells. Thus, gel containing EA can be tested as a new microbicide against HIV infection.
鞣花酸（EA）具有广泛的生物学效应。本研究旨在探讨 EA 在体外对 HIV-1 复制、病毒酶活性和感染细胞细胞因子分泌的影响。
采用纳米荧光素酶分泌的 R5 嗜性 JRCSF 株 HIV-1 感染 TZM-bl 细胞，通过测量培养上清液中的纳米荧光素酶来定量病毒生长，在体外测定 EA 在溶液中的抗 HIV-1 活性。在 HIV-1 暴露 48 小时后，通过多指标 bead 阵列测定 EA 对 TZM-bl 细胞细胞因子分泌的影响。采用 Ellagel（凝胶制剂）抑制 R5 嗜性 pBaLEnv-重组 HIV-1 感染 UC87.CD4.CCR5 细胞，研究 EA 在凝胶制剂中的抗病毒作用。
浓度高达 100 μM 的 EA 对 TZM-bl 细胞无毒性。在 HIV-1 暴露前 1 小时或后 4 小时加入 EA，可剂量依赖性地抑制 TZM-bl 细胞中 R5 嗜性 HIV-1 的复制，在 50 μM 时最高抑制率为 69%。含 EA 的溶液也表现出对 U87 细胞中 HIV-1 复制的剂量依赖性抑制作用。当 EA 被制成凝胶时，含有 25 μM 和 50 μM EA 的 Ellagel 分别抑制 U87 细胞中 HIV-1 的复制 56%和 84%。在特定 HIV-1 酶活性测定中，Ellagel 抑制 HIV-1 整合酶，但不抑制蛋白酶。EA 对细胞因子分泌没有显著调节作用。
我们得出结论，EA 无论是在溶液中还是在凝胶形式下，都能抑制 HIV 感染而不对靶细胞产生不良反应。因此，含有 EA 的凝胶可以作为一种新的抗 HIV 感染的杀微生物剂进行测试。
