南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (12): 1693-1702.doi: 10.12122/j.issn.1673-4254.2020.12.01

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SBi4211通过抑制S100B/RAGE表达减轻gp120诱导的中枢神经损伤

杨少杰，邓小燕，张铁松，肖 仪，彭 亮，李 莉，何肖龙，魏 轶，刘利群，曹 虹，龙北国，黄胜和   

  • 出版日期:2020-12-20 发布日期:2020-12-29

SBi4211 alleviates gp120-induced central nervous system injury via inhibiting S100B/RAGE

  • Online:2020-12-20 Published:2020-12-29

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摘要： 目的 研究S100B 抑制剂SBi4211（heptamidine）在人类免疫缺陷病毒-1（HIV-1）包膜蛋白gp120损伤中枢神经系统的作用。方法 通过U251细胞和SH-SY5Y细胞建立非接触式星形胶质细胞-神经元共培养体系，由gp120蛋白处理U251细胞激活神经炎症反应造成神经元损伤，体外水平探讨SBi4211在gp120诱导的中枢神经毒性中的作用；体内实验中，以8月龄gp120转基因小鼠模拟HIV相关神经认知障碍（HAND）模型，实验分为对照组、gp120组以及gp120+SBi4211组（SBi4211处理组）。采用CCK-8、流式细胞术检测神经元活性及凋亡情况，ELISA检测S100B及炎症因子IL-6、TNF-α表达水平，Western blot和免疫组织化学染色分析RAGE、GFAP、NeuN、Syn、MAP-2蛋白表达情况。结果 体外实验结果显示SBi4211可以显著抑制gp120刺激后U251细胞S100B、RAGE的表达（P<0.001），降低炎症因子iNOS、IL-6和TNF-α的表达水平（P<0.001），维持神经元相关标记蛋白NeuN、Syn的表达（P<0.001）。体内实验结果显示SBi4211显著降低gp120转基因小鼠S100B、RAGE表达及炎症水平（P<0.05），并且抑制脑内星形胶质细胞活化，保护神经元的完整性（P<0.05）。结论 SBi4211可能通过下调S100B/RAGE表达，抑制gp120引发的神经炎症反应，继而阻断gp120的中枢神经毒性作用。

关键词: HIV-1 gp120；HAND；SBi4211；S100B/RAGE

Abstract: Objective To explore the protective effect of SBi4211 (heptamidine), an inhibitor of S100B, against central nervous system injury induced by HIV-1 envelope protein gp120. Methods In an in vitro model, U251 glioma cells were co-cultured with SH-SY5Y cells to explore the protective effect of SBi4211 against gp120-induced central nervous system injury. In a gp120 transgenic (Tg) mouse model (8 months old) mimicking HIV-associated neurocognitive disorder (HAND), the effect of treatment with gp120 or both gp120 and SBi4211 on neuronal activity and apoptosis were assessed using Cell Counting kit-8 (CCK-8) and flow cytometry. ELISA, Western blotting and immunohistochemistry were used to determine the expression levels of S100B, RAGE, GFAP, NeuN, Syn, MAP-2 and the inflammatory factors IL-6 and TNF-α. Results In the cell co-culture system, SBi4211 treatment significantly inhibited gp120-induced expression of S100B, RAGE and GFAP in U251 cells (P<0.001), reduced the levels of inflammatory factors iNOS, IL-6 and TNF-α (P<0.001) and enhanced the expressions of neuron-related proteins NeuN, Syn and MAP-2 (P<0.001). In the transgenic mouse model, SBi4211 treatment significantly reduced the expressions of S100B, RAGE and inflammation levels (P<0.05), inhibited the activation of astrocytes in the brain, and maintained the integrity of the neurons (P<0.05). Conclusion SBi4211 can protect neurons from gp120-induced neurotoxicity possibly by inhibiting the S100B/RAGE-mediated signaling pathway

Key words: gp120; HIV-associated neurocognitive disorder; SBi4211; S100B/RAGE