过表达PGRN抑制LPS诱导的肺泡上皮细胞损伤

发布时间：2018-04-18 23:00

本文选题：颗粒体蛋白前体 + 脂多糖　；参考：《中国病理生理杂志》2017年05期

【摘要】：目的:探讨颗粒体蛋白前体(progranulin,PGRN)对脂多糖(lipopolysaccharide,LPS)诱导的人肺泡上皮A549细胞和HPAEpi C细胞增殖、凋亡及炎症的影响。方法:实验分为4组:对照(control)组为正常培养的细胞;LPS组用LPS(10 mg/L)处理;PGRN+LPS组在转染pc DNA3.1-PGRN质粒后加入LPS处理;pc DNA3.1+LPS组在转染pc DNA3.1-EGFP质粒后加入LPS处理。MTT试剂盒检测细胞活力;Brd U掺入实验检测细胞增殖;流式细胞术检测细胞凋亡率;RT-q PCR和Western blot分别检测PGRN的mRNA和蛋白表达;Western blot检测caspase-3、Bcl-2、Bax、IL-1β、IL-6、TNF-α、p65和p-IκB-α的蛋白水平。结果:与对照组相比,LPS组的细胞增殖率下降(P0.05),凋亡率上升(P0.05),caspase-3和Bax的蛋白表达上调(P0.05),Bcl-2的蛋白表达下调(P0.05),IL-1β、IL-6和TNF-α的蛋白表达上调(P0.05),p65和p-IκB-α的蛋白水平增加(P0.05),与LPS组相比,PGRN+LPS组的细胞增殖率上升(P0.05),凋亡率下降(P0.05),caspase-3和Bax的蛋白表达下调(P0.05),Bcl-2的蛋白表达上调(P0.05),IL-1β、IL-6和TNF-α的蛋白表达下调(P0.05),p65和p-IκB-α的蛋白水平降低(P0.05)。结论:PGRN过表达可以减轻LPS诱导的A549细胞和HPAEpi C细胞增殖、凋亡异常及炎症因子产生等损伤,这可能与PGRN参与调控NF-κB信号通路有关。
[Abstract]:Aim: to investigate the effects of progranulinine progranulinone (PGRN) on the proliferation, apoptosis and inflammation of human alveolar epithelial A549 cells and HPAEpi C cells induced by lipopolysaccharide (LPS).Methods: the experiment was divided into four groups: control control group was treated with LPS(10 mg / L, the control group was treated with LPS(10 mg / L, and the control group was treated with LPS after transfection of PC DNA3.1-PGRN plasmid. The control group was transfected with PC DNA3.1-EGFP plasmid and LPS treatment. MTT kit was added to detect the cells in the control group after transfection of PC DNA3.1-EGFP plasmid.Cell proliferation was detected by BrdU incorporation assay.Apoptosis rate was detected by flow cytometry. MRNA and protein expression of PGRN were detected by RT-q PCR and Western blot respectively. Western blot was used to detect the protein levels of caspase-3, Bcl-2Baxan-1 尾, IL-6, TNF- 伪, p65 and p-I 魏 B- 伪.Results: compared with the control group, the cell proliferation rate of lipopolysaccharide group decreased and the apoptosis rate increased. The protein expression of p0.05 caspase-3 and Bax was up-regulated. The protein expression of P0.05, IL-1 尾, IL-6 and TNF- 伪 was down-regulated. Compared with LPS group, the expression of p65 and p-I 魏 B- 伪 increased, and the protein levels of P0.05 and p-I 魏 B- 伪 increased.In LPS group, the cell proliferation rate increased and the apoptosis rate decreased. The protein expression of P0.05 caspase-3 and Bax down-regulated the protein expression of P0.05, IL-1 尾, IL-6 and TNF- 伪, and down-regulated the protein levels of p0.05, p65 and p-I 魏 B- 伪, and decreased the protein levels of P0.05 and p-I 魏 B- 伪.Conclusion the overexpression of WPGRN can attenuate the proliferation, abnormal apoptosis and the production of inflammatory factors in A549 cells and HPAEpi C cells induced by LPS, which may be related to the regulation of NF- 魏 B signaling pathway by PGRN.
【作者单位】：陕西省人民医院呼吸内一科;
【分类号】：R563.1

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