FAM134B介导的内质网自噬对脓毒症的影响

Effect of FAM134B mediated ER-phagy in sepsis

摘要:

目的 初步探讨FAM134B介导的内质网自噬在脓毒症中作用。方法 采用盲肠结扎穿孔术（CLP）复制脓毒症大鼠模型，取脑组织后，qRT-PCR检测mRNA水平、Western blotting检测FAM134B蛋白表达量；在脂多糖（LPS）诱导的N2a脓毒症细胞模型中，免疫荧光检测FAM134B在细胞中的分布，Western blotting检测自噬相关蛋白LC3，Beclin1及凋亡相关蛋白活化Caspase-3的表达；Annexin V-FITC/PI双染流式细胞仪分析检测细胞在不同处理后凋亡率；过表达FAM134B细胞中，LPS处理后分别检测自噬及凋亡相关蛋白的表达及细胞凋亡情况。结果 与正常对照组比较，脓毒症大鼠脑组织中内质网蛋白FAM134B表达下降（P?<0.05）；LPS诱导脓毒症细胞模型中内质网自噬及细胞凋亡，且过表达FAM134B能减少LPS对细胞的损害（P?<0.05）。结论 FAM134B介导的内质网自噬影响脓毒症的发生。

Abstract:

Objective To investigate the role of FAM134B-mediated ER-phagy in the pathogenesis of sepsis. Methods A rat model of sepsis was constructed by Cecal Ligation and Puncture (CLP). The brain expression levels of FAM134B was detected by qRT-PCR and Western Blotting. In LPS induced N2a sepsis cell model, immunofluorescence staining was performed for the distribution of FAM134B in cells. The expression level of autophagy-associated protein LC3, Beclin1 and apoptosis-related protein Caspase-3 were detected by Western blotting. The rate of apoptosis in each group was measured by Annexin V-FITC/PI flow cytometry. Also, the expression levels of autophagy and apoptosis-related proteins were detected in FAM134B over-expression cell lines after LPS treatment. Results The expression of FAM134B in the septic brain were decreased. LPS insulted endoplasmic reticulum autophagy and cell apoptosis were increased. FAM134B overexpression reduced LPS induced damage to cells. Conclusions FAM134B mediated ER-phagy plays a protective role in sepsis.