大鼠高迁移率族蛋白B1表达水平与COPD肺动脉平滑肌细胞合成分泌干扰素-γ的关系

发布时间：2018-06-18 10:26

本文选题：高迁移率族蛋白B + 慢性阻塞性肺疾病　；参考：《医学研究生学报》2014年12期

【摘要】：目的肺动脉高压与慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)密切相关。研究COPD继发肺动脉高压的机制,探讨COPD模型大鼠远端肺动脉平滑肌细胞(pulmonary arterial smooth muscle cells,PASMCs)高迁移率族蛋白B1(high mobility group protein B1,HMGB1)表达水平与其合成分泌干扰素-γ(interferon-γ,IFN-γ)的相关性及临床意义。方法建立COPD大鼠模型,原代培养PASMCs并鉴定,检测香烟烟雾提取物(CSE)及脂多糖(LPS)诱导下PASMCs合成分泌细胞因子的功能,实验分组:对照组、CSE组、LPS组、CSE+LPS组,同时4组另设瓶并分别加入1∶1000抗HMGB1抗体0.2 m L培养作为干预组;分别于培养12、24、48、72 h后,采用Western blot检测各组PASMCs中HMGB1蛋白表达变化,ELISA检测细胞培养上清液HMGB1、IFN-γ浓度,并进行相关分析。结果光镜下见细胞呈长梭形或"峰、谷"状生长,α-actin免疫组化染色及细胞免疫荧光鉴定为PASMCs;Western blot及ELISA法检测结果显示,4实验组组间培养12 h后HMGB1表达水平及细胞上清液HMGB1、IFN-γ,两两比较差异均有统计学意义(P0.05);HMGB1蛋白表达水平与IFN-γ浓度变化呈正相关(r=0.91);加入抗HMGB1抗体干预培养12 h后,干预组IFN-γ浓度均较对应实验组减弱(P0.05);同组内,于细胞培养12、24、48、72 h,Western blot及ELISA法检测结果差异均无统计学意义(P0.05)。结论 HMGB1表达水平与COPD模型大鼠远端PASMCs合成分泌IFN-γ呈正相关,抗HMGB1抗体为临床治疗COPD、抑制其全身炎症反应提供了新的干预靶点。
[Abstract]:Objective Pulmonary hypertension is closely related to chronic obstructive pulmonary disease (obstructive pulmonary). To investigate the mechanism of pulmonary hypertension, and to investigate the relationship between the expression of high mobility group protein B1 (high mobility group protein B1HMGB1) and the synthesis and secretion of interferon- 纬 -interferon- 纬 (IFN- 纬) in pulmonary smooth muscle cells of distal pulmonary artery of COPD model rats. Methods the rat model of COPD was established, PASMCs were cultured in vitro and identified. The synthesis and secretion of cytokines of PASMCs induced by cigarette smoke extract (CSE) and lipopolysaccharide (LPSs) were detected. The rats were divided into two groups: control group (CSE group) and LPS group (LPS group). At the same time, the 4 groups were cultured with 1: 1000 anti-HMGB1 antibody 0.2 mL respectively as the intervention group, the expression of HMGB1 protein in PASMCs of each group was detected by Western blot and the concentration of HMGB1 IFN- 纬 in the supernatant of cell culture was detected by Elisa. And carry on the correlation analysis. Results under the light microscope, the cells appeared to be fusiform or "peak". The expression of HMGB1 and the expression of HMGB1 in the supernatant of HMGB1 and HMGB1 in cell supernatant were detected by Western blot and Elisa after 12 h culture. There were significant differences between the two groups in the expression of HMGB1 and the expression of HMGB1 in the supernatant of HMGB1. The expression of HMGB1 and the expression of HMGB1 in the cell supernatant were significantly higher than those in the control group. The level of protein expression was positively correlated with the change of IFN- 纬 concentration, and the anti-HMGB1 antibody was added to culture for 12 h. The concentration of IFN- 纬 in the intervention group was lower than that in the corresponding experimental group, while in the same group, there was no significant difference in the results of Western blot and Elisa in the same group. Conclusion the level of HMGB1 expression is positively correlated with the synthesis and secretion of IFN- 纬 by distal PASMCs in COPD model rats. Anti-HMGB1 antibody provides a new intervention target for clinical treatment of COPDs and inhibition of systemic inflammatory response.
【作者单位】：桂林医学院附属医院呼吸内科;
【基金】：国家自然科学基金(81360010) 广西壮族自治区医疗卫生重点科研课题(2012004)
【分类号】：R563.9

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