热休克蛋白A12B对脂多糖诱导肺微血管内皮细胞损伤的影响
发布时间：2018-08-01 08:06
【摘要】：目的探讨小鼠肺微血管内皮细胞(mPMVECs)热休克蛋白A12B(HSPA12B)表达下调对脂多糖(LPS)诱导的细胞炎症反应、迁移及超微结构改变的影响。方法体外传代培养的mPMVECs分为LPS组(添加1μg/mL LPS)、LPS+siRNA组[瞬时转染法将HSPA12B相应基因片段干扰小RNA(siRNA)寡核苷酸转入mPMVECs中,再加入1μg/mL LPS)]和LPS+NC组[瞬时转染法将平行对照(NC)siRNA寡核苷酸转入mPMVECs中,再加入1μg/mL LPS]。对LPS+siRNA组和LPS+NC组,分别采用Transwell试验和细胞划痕实验观察细胞迁移情况;应用透射电子显微镜(透射电镜)观察mPMVECs超微结构改变;应用Real-Time PCR检测各组细胞中肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、IL-10和IL-1βmRNA的表达。结果与LPS+NC组比较,LPS+siRNA组mPMVECs的线粒体损伤加重,内质网水肿更明显;细胞内TNF-α、IL-6和IL-10mRNA的表达显著上调(P0.05),而IL-1β表达下调,但差异无统计学意义(P0.05);细胞迁移功能显著下降(P0.05)。结论下调HSPA12B表达后,mPMVECs经LPS刺激的炎症反应增强,细胞迁移功能下降。HSPA12B可能参与保护mPMVECs免受LPS侵袭的过程。
[Abstract]:Objective to investigate the effect of down-regulation of (mPMVECs) heat shock protein A12B (HSPA12B) expression in mouse pulmonary microvascular endothelial cells on cellular inflammation, migration and ultrastructural changes induced by lipopolysaccharide (LPS). Methods mPMVECs cultured in vitro was divided into LPS group (1 渭 g/mL LPS) and LPS siRNA group [transient transfection of HSPA12B gene fragment interfering small RNA (siRNA) oligonucleotide into mPMVECs. In the LPS NC group, parallel control (NC) siRNA oligonucleotides were transferred into mPMVECs by transient transfection and then 1 渭 g/mL LPS was added. In LPS siRNA group and LPS NC group, cell migration was observed by Transwell test and cell scratch test, and ultrastructural changes of mPMVECs were observed by transmission electron microscope (TEM). The expression of tumor necrosis factor- 伪 (TNF- 伪), interleukin-6 (IL-6) IL-10 and IL-1 尾 mRNA were detected by Real-Time PCR. Results compared with LPS NC group, the mitochondria damage and endoplasmic reticulum edema were more serious in mPMVECs group than those in LPS NC group, the expression of TNF- 伪 IL-6 and IL-10mRNA was significantly up-regulated (P0.05), but the expression of IL-1 尾 was down-regulated, but the difference was not statistically significant (P0.05), and the cell migration function was significantly decreased (P0.05). Conclusion after down-regulation of HSPA12B expression, the inflammatory response of mPMVECs stimulated by LPS is enhanced, and the cell migration function is decreased. HSPA12B may be involved in the process of protecting mPMVECs from LPS invasion.
【作者单位】： 第二军医大学长海医院麻醉科;上海交通大学医学院附属第三人民医院麻醉科;
【分类号】：R563
，
本文编号：2156856
[Abstract]:Objective to investigate the effect of down-regulation of (mPMVECs) heat shock protein A12B (HSPA12B) expression in mouse pulmonary microvascular endothelial cells on cellular inflammation, migration and ultrastructural changes induced by lipopolysaccharide (LPS). Methods mPMVECs cultured in vitro was divided into LPS group (1 渭 g/mL LPS) and LPS siRNA group [transient transfection of HSPA12B gene fragment interfering small RNA (siRNA) oligonucleotide into mPMVECs. In the LPS NC group, parallel control (NC) siRNA oligonucleotides were transferred into mPMVECs by transient transfection and then 1 渭 g/mL LPS was added. In LPS siRNA group and LPS NC group, cell migration was observed by Transwell test and cell scratch test, and ultrastructural changes of mPMVECs were observed by transmission electron microscope (TEM). The expression of tumor necrosis factor- 伪 (TNF- 伪), interleukin-6 (IL-6) IL-10 and IL-1 尾 mRNA were detected by Real-Time PCR. Results compared with LPS NC group, the mitochondria damage and endoplasmic reticulum edema were more serious in mPMVECs group than those in LPS NC group, the expression of TNF- 伪 IL-6 and IL-10mRNA was significantly up-regulated (P0.05), but the expression of IL-1 尾 was down-regulated, but the difference was not statistically significant (P0.05), and the cell migration function was significantly decreased (P0.05). Conclusion after down-regulation of HSPA12B expression, the inflammatory response of mPMVECs stimulated by LPS is enhanced, and the cell migration function is decreased. HSPA12B may be involved in the process of protecting mPMVECs from LPS invasion.
【作者单位】： 第二军医大学长海医院麻醉科;上海交通大学医学院附属第三人民医院麻醉科;
【分类号】：R563
，
本文编号：2156856
本文链接：https://www.wllwen.com/yixuelunwen/huxijib/2156856.html
最近更新
教材专著
