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

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／mLLPS）、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／mLLPS）]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／mLLPS]。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...

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Published in上海交通大学学报：医学版 Vol. 34; no. 7; pp. 1022 - 1026
Main Author 余桂芳 张旭 朱科明
Format Journal Article
LanguageChinese
Published 2014
Subjects
小鼠肺微血管内皮细胞
热休克蛋白A12B
脂多糖
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Summary: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／mLLPS）、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／mLLPS）]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／mLLPS]。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-TimePCR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（P〈0．05），而IL-1β表达下调，但差异无统计学意义（P〉0．05）；细胞迁移功能显著下降（P〈0．05）。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。
Bibliography:YU Gui-fang , ZHANG Xu , ZHU Ke-ming（ 1. Department of Anesthesiology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China; 2. Department of Anesthesiology, the Third People＇s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201999, China）
heat shock protein A12B; lipopolysaccharide; mouse pulmonary microvascular endothelia cells
31-1259/R
Objective To explore the effects of down-regulation of the expression of heat shock protein A12B （HSPA12B） of mouse pulmonary microvascular endothefia cells （mPMVECs） on the inflammatory reaction, migration, and ultrastructural changes induced by fipopolysaccharide （LPS）. Methods The mPMVECs subcultured in vitro were divided into the LPS group （LPS of 1 μg/mL was added）, LPS ＋ siRNA group （siRNA oligonucleotides of relevant gene segments of HSPA12B were transiently transfected into mPMVECs and then LPS of 1 μg/mL was added）, and LPS ＋ NC group （siRNA oligonucleotides of negative controls were transiently transfected into mPMVECs an
ISSN:1674-8115