Yang Jie, Li Lin, Jin Hong, Tan Suiyi, Qiu Jiayin, Yang Lei, Ding Yanqing, Jiang Zhi-Hong, Jiang Shibo, Liu Shuwen
School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, China.
AIDS Res Hum Retroviruses. 2012 Nov;28(11):1498-508. doi: 10.1089/AID.2012.0084. Epub 2012 Sep 11.
We previously demonstrated that a commercially available natural product preparation with high content (>90%) of theaflavin derivatives (TFmix) exhibited potent anti-HIV activities. Here we developed a TFmix gel formulation as a topical microbicide candidate. The effect of TFmix on the amyloid fibril formation of semen-derived enhancer of virus infection (SEVI) peptide was detected by transmission electron microscopy. The toxicity of the TFmix gel was evaluated using human vaginal and cervical epithelial cell lines and rabbit vaginal irritation models, respectively. Levels of proinflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α) and immunoregulatory cytokines (IL-10 and GM-CSF) in cervicovaginal lavages (CVLs) were measured by ELISA kits. Proliferating cell nuclear antigen (PCNA) immunostaining was performed to evaluate inflammation in the vaginal tissues. TFmix gel could degrade SEVI-specific amyloid fibrils and showed low cytotoxicity to epithelial cells of the female reproductive tract. No apparent cervicovaginal toxicity was observed at any time point evaluated following the intravaginal administration of TFmix gel to rabbits, whereas application of N-9 gel resulted in damage to the vaginal epithelium. Neither proinflammatory nor immunoregulatory cytokine production was triggered by TFmix gel. Only low expression of PCNA was observed in vaginal tissues of TFmix gel-treated rabbits. The concentration of TFmix in plasma was very low (below the lower limit of quantitation) 1 h after a single vaginal administration of TFmix gel. However, TFmix was still detected in the cervicovaginal lavages (CVLs) 6 h after treatment, indicating that it could be retained in the vaginal cavity for a long period of time. With its potent anti-HIV-1 activity, marked stability at acidic condition, low mucosal toxicity, and lack of systemic absorption, TFmix gel can be considered as an inexpensive and safe microbicide candidate for the prevention of HIV sexual transmission.
我们之前证明，一种市售的、茶黄素衍生物含量高（>90%）的天然产物制剂（TFmix）具有强大的抗HIV活性。在此，我们开发了一种TFmix凝胶制剂作为局部杀菌剂候选物。通过透射电子显微镜检测TFmix对精液衍生病毒感染增强剂（SEVI）肽淀粉样原纤维形成的影响。分别使用人阴道和宫颈上皮细胞系以及兔阴道刺激模型评估TFmix凝胶的毒性。通过酶联免疫吸附测定试剂盒测量宫颈阴道灌洗液（CVL）中促炎细胞因子（IL-1β、IL-6、IL-8和TNF-α）和免疫调节细胞因子（IL-10和GM-CSF）的水平。进行增殖细胞核抗原（PCNA）免疫染色以评估阴道组织中的炎症。TFmix凝胶可降解SEVI特异性淀粉样原纤维，对女性生殖道上皮细胞显示出低细胞毒性。对兔阴道内给予TFmix凝胶后，在评估的任何时间点均未观察到明显的宫颈阴道毒性，而应用N-9凝胶则导致阴道上皮损伤。TFmix凝胶未引发促炎或免疫调节细胞因子的产生。在TFmix凝胶处理的兔阴道组织中仅观察到PCNA的低表达。单次阴道给予TFmix凝胶1小时后，血浆中TFmix的浓度非常低（低于定量下限）。然而，治疗6小时后在宫颈阴道灌洗液（CVL）中仍检测到TFmix，表明它可以在阴道腔内长时间保留。凭借其强大的抗HIV-1活性、在酸性条件下的显著稳定性、低黏膜毒性以及缺乏全身吸收，TFmix凝胶可被视为一种用于预防HIV性传播的廉价且安全的杀菌剂候选物。
