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目的探讨游离胆固醇激活未折叠蛋白反应以及对巨噬细胞凋亡的影响。方法取小鼠腹腔巨噬细胞进行培养,然后分别以普通培养基、普通培养基加胆固醇乙酰转移酶抑制剂和乙酰低密度脂蛋白、普通培养基加胆固醇乙酰转移酶抑制剂和乙酰低密度脂蛋白再加胆固醇移动抑制剂分组孵育,Annexin-V和碘化丙锭双染色后流式细胞仪检测细胞凋亡,Western-bolt检测下游转录因子C/EBP同源蛋白表达。结果普通培养基孵育的巨噬细胞无C/EBP同源蛋白表达,只有少量细胞凋亡;而普通培养基加胆固醇乙酰转移酶抑制剂和乙酰低密度脂蛋白孵育的巨噬细胞C/EBP同源蛋白表达明显,细胞凋亡也达到21.83%±2.47%;同时使用胆固醇移动抑制剂干预后,C/EBP同源蛋白无表达,而细胞凋亡比单纯促进游离胆固醇聚集状态孵育组显著减少。结论过多的游离胆固醇聚集是诱导巨噬细胞凋亡的重要原因之一,未折叠蛋白反应参与这一过程,而特异性胆固醇移动抑制剂U18666A可以有效阻止游离胆固醇进入内质网激活未折叠蛋白反应诱导巨噬细胞凋亡。
Objective To investigate the effects of free cholesterol-activated unfolded protein and its effect on macrophage apoptosis. Methods Mice peritoneal macrophages were cultured and then cultured in normal medium, normal medium plus cholesterol acetyltransferase inhibitors and acetyl low-density lipoprotein, ordinary medium plus cholesterol acetyltransferase inhibitors and acetyl low-density lipids Protein plus cholesterol mobilization inhibitor grouping, Annexin-V and propidium iodide double staining were used to detect apoptosis by flow cytometry. Western-blot was used to detect the expression of downstream transcription factor C / EBP homology protein. Results There was no C / EBP homologous protein expression in macrophages cultured in normal medium, but only a small amount of apoptotic cells. C / EBP of macrophages cultured with normal medium plus cholesterol acetyltransferase inhibitor and acetyl-LDL The expression of C / EBP homology protein was significantly lower than that of the control group (21.83% ± 2.47%). In the meantime, C / EBP homologous protein was not expressed after intervention with cholesterol mobilization inhibitor. Conclusion Excessive free cholesterol accumulation is one of the important reasons for inducing macrophage apoptosis. Unfolded protein responses are involved in this process. The specific cholesterol transport inhibitor U18666A can effectively prevent free cholesterol from entering the endoplasmic reticulum to activate unfolded proteins Reactions induce macrophage apoptosis.