Ras信号通路在HIV-1 Tat诱导ZO-1及脑啡肽酶破坏的作用

发布时间：2018-07-06 21:33

本文选题：HIV- + Tat　；参考：《中风与神经疾病杂志》2017年02期

【摘要】：目的探讨Ras信号传导通路在人类免疫缺陷病毒-1型反式转录激活因子(HIV-1 transactivator of transcription,HIV-1 Tat)诱导血脑屏障(BBB)中紧密连接蛋白Zonula Occludens(ZO)-1及脑啡肽酶(Neprilysin,NEP)破坏的作用。方法以不同浓度Ras信号传导通路抑制剂法尼基硫代水杨酸(farnesylthiosalicylic acid,FTS)刺激培养好的人脑微血管内皮细胞(human cerebral microvascular endothelium cells,HBEC-5i),并设立对照组,观察其对细胞活力的影响。分别予以HIV-1 Tat、FTS刺激细胞,以蛋白免疫印迹法及实时反转录聚合酶链式反应(realtime reverse transcription polymerase chain reaction,RT-PCR)检测HIV-1 Tat诱导的紧密连接蛋白ZO-1及NEP(脑内降解β淀粉样蛋白Amyloid-beta,Aβ的限速酶)的蛋白和mRNA表达的变化。结果 FTS在20μmol/L(0.35±0.06)以下时对HBEC-5i活力无明显影响(P0.05),HIV-1 Tat能抑制HBEC-5i的ZO-1、NEP蛋白(0.53±0.07,P0.01;0.40±0.04,P0.05)与mRNA(0.42±0.10,P0.05;0.31±0.09,P0.05)的表达。用FTS阻断Ras信号通路后可显著增加ZO-1、NEP蛋白(1.20±0.22,P0.01;0.58±0.03,P0.05)及mRNA(1.10±0.19,P0.05;0.97±0.38,P0.05)的表达。结论 HIV-1 Tat可促进紧密连接蛋白ZO-1蛋白和mRNA下调而导致血脑屏障破坏,同时诱导脑内脑啡肽酶NEP蛋白和mRNA下调,可能导致Aβ在脑内沉积增加。阻断Ras信号传导通路可抑制HIV-1 Tat诱导的ZO-1和NEP破坏,可能减少Aβ在脑内的沉积。
[Abstract]:Objective to investigate the effect of Ras signal transduction pathway on the destruction of tight junction proteins Zonula Occludens (ZO) -1 and neprilysin nep (NEP) in blood brain barrier (BBB) induced by HIV-1 transactivator of transcriptional activator (HIV-1 tat). Methods the cultured human microvascular endothelial cells (human cerebral microvascular endothelium cells) were stimulated with different concentrations of Ras signal transduction pathway inhibitor farnesylthiosalicylic acidosalicylic acid (farnesylthiosalicylic acidosalicylic acid) and the control group was set up to observe its effect on cell viability. The cells were stimulated by HIV-1 Taton FTS. The protein and mRNA expression of tight junction protein ZO-1 and nep (rate-limiting enzyme for degradation of amyloid beta A 尾) induced by HIV-1 tat were detected by Western blot and real-time reverse transcription polymerase chain reaction (RT-PCR). Results when the concentration of FTS was below 20 渭 mol / L (0.35 卤0.06), there was no significant effect on the activity of HBEC-5i (P0.05). HIV-1 tat could inhibit the expression of ZO-1NEP protein (0.53 卤0.07) and mRNA (0.42 卤0.10P0.05N) and mRNA (0.42 卤0.10P0.05P, 0.31 卤0.09p0.05) of HBEC-5i. After blocking the Ras signal pathway with FTS, the expression of ZO-1Nep protein (1.20 卤0.22P0.01U 0.58 卤0.03p0.05) and mRNA (1.10 卤0.19P0.0550.97 卤0.38p0.05) were significantly increased. Conclusion HIV-1 tat can promote the down-regulation of tight junction protein ZO-1 protein and mRNA leading to the breakdown of blood-brain barrier, and induce the down-regulation of NEP protein and mRNA in the brain, which may lead to the increase of A 尾 deposition in the brain. Blocking the Ras signaling pathway can inhibit the destruction of ZO-1 and NEP induced by HIV-1 tat, and may reduce the deposition of A 尾 in the brain.
【作者单位】：广西医科大学第一附属医院神经内科;广西广西壮族自治区南溪山医院神经内科;
【基金】：国家自然科学基金(No.81371333)
【分类号】：R512.91;R747.9

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