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目的通过研究灭活HIV-1IIIB颗粒对人结肠癌HT-29细胞的增殖、凋亡和紧密连接的影响,探讨HIV-1病毒子在“AIDS肠道病灶论”中的直接作用机制。方法将AT-2灭活的HIV-1IIIB颗粒加入到HT-29培养系统中,使p24抗原终质量浓度为1 ng/ml;WST-1法检测HT-29的增殖率;运用流式细胞术检测HT-29中Ki-67和Apo2.7的表达以及线粒体膜电势和线粒体质量的变化;运用流式细胞术检测HT-29紧密连接蛋白Occludin的表达。结果灭活HIV-1IIIB抑制HT-29增殖并抑制Ki-67表达;灭活HIV-1IIIB诱导HT-29凋亡并降低线粒体膜电势和线粒体质量;灭活HIV-1IIIB下调Occludin表达,提示其破坏细胞的紧密连接。结论 AT-2灭活HIV-1IIIB能够抑制HT-29增殖,诱导HT-29凋亡,并破坏HT-29紧密连接,提示在“AIDS肠道病灶论”中,导致肠黏膜屏障完整性受损的因素除了HIV-1引发的“旁观者效应”外,HIV-1病毒子本身可以对肠上皮细胞(intestinal epithelial cell,IEC)产生细胞毒作用;而肠黏膜通透性增加会引起肠菌转位,继而引发系统性免疫过度活化。
OBJECTIVE: To investigate the direct mechanism of HIV-1 virion in the development of “gastrointestinal lesions” by studying the effect of inactivated HIV-1IIIB particles on the proliferation, apoptosis and tight junction of human colon cancer HT-29 cells. Methods AT-2 inactivated HIV-1IIIB particles were added into the HT-29 culture system to make the final concentration of p24 antigen 1 ng / ml. The proliferation rate of HT-29 was detected by WST-1 assay. Flow cytometry The expression of Ki-67 and Apo2.7 in HT-29 and the changes of mitochondrial membrane potential and mitochondrial mass were detected. The expression of tight junction protein Occludin in HT-29 was detected by flow cytometry. Results Inactivation of HIV-1IIIB inhibited HT-29 proliferation and inhibited Ki-67 expression. Inactivation of HIV-1IIIB induced HT-29 apoptosis and decreased mitochondrial membrane potential and mitochondrial mass. Inactivation of HIV-1IIIB down-regulated Occludin expression, Close connection of cells. Conclusions AT-2 inactivation of HIV-1IIIB can inhibit the proliferation of HT-29, induce the apoptosis of HT-29, and destroy the tight junctions of HT-29, suggesting that the intestinal mucosal barrier integrity Impaired Factor In addition to the “bystander effect” triggered by HIV-1, the HIV-1 virion itself can exert cytotoxic effects on the intestinal epithelial cells (IEC), whereas increased intestinal mucosal permeability Causing enterotrophic translocation, which in turn lead to systemic immune over-activation.