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山东大学学报(医学版) ›› 2013, Vol. 51 ›› Issue (5): 58-61.

• 基础医学 • 上一篇    下一篇

未折叠蛋白反应在严重烧伤大鼠淋巴液诱导肺内皮细胞凋亡中的作用

唐公杰1,王德昌2,薛文君2,张涛1,赵春平1   

  1. 1.沂水中心医院烧伤整形科, 山东 沂水 276400; 2.山东大学附属省立医院烧伤整形科, 济南 250021
  • 收稿日期:2012-09-10 出版日期:2013-05-10 发布日期:2013-05-10
  • 通讯作者: 王德昌, E-mail:wangdechang0531@126.com

Lymph of rats with severe scalds induces apoptosis in vascular endothelial cell via the unfolded protein response

TANG Gong-jie1, WANG De-chang2, XUE Wen-jun2, ZHANG Tao1, ZHAO Chun-ping1   

  1. 1. Department of Scalds and Plastic Surgery, Yishui Hospital, Yishui 276400, Shandong, China;
    2. Department of Scalds and Plastic Surgery, Provincial Hospital Affiliated to Shandong University, Jinan 250021, China
  • Received:2012-09-10 Online:2013-05-10 Published:2013-05-10

摘要:

目的   探讨未折叠蛋白反应介导的凋亡通路在严重烧伤大鼠淋巴液对大鼠肺微血管内皮细胞损伤中的作用。方法   选成年Wistar雄性大鼠20只,分为烫伤组(行30%体表面积Ⅲ度烫伤)和假伤组,建立大鼠胸导管淋巴瘘模型,收集伤后24h淋巴液,用于体外实验。将细胞实验分为烫伤淋巴液刺激组(A组)、烫伤淋巴液+4苯基丁酸(4PBA)处理组(B组)和假伤淋巴液处理组(C组)。刺激6h后,Annexin V/PI标记法流式细胞术检测细胞凋亡。western blot检测caspase12、葡萄糖调节蛋白78 (GRP78)和C/EBP同源蛋白(CHOP)等蛋白表达的情况。结果    A组细胞凋亡水平显著高于C组(P<0.01),B组细胞凋亡水平显著低于A组(P<0.01)。A组GRP78、CHOP和caspase12蛋白表达水平显著高于C组,经PBA处理后3种蛋白水平显著降低(P<0.01)。结论   烧伤大鼠淋巴液可以引起血管内皮细胞凋亡,未折叠蛋白反应可能是其主要通路之一。

关键词: 淋巴液;内皮细胞;未折叠蛋白反应;凋亡;肺损伤

Abstract:

Objective   To investigate the effects of the unfolded protein response(UPR) in endoplasmic reticulum stress on the apoptosis of rat pulmonary microvascular endothelial cells(rPMECs) induced by lymph from rats with severe scalds. Methods   Twenty Wistar rats were divided into burn group(n=10) and sham burn group(n=10). Lymph was harvested within 24 hours after the establishment of scalds model. Cultured rPMECs were divided into scald lymph stimulated group (A,stimulated by 5% scalded rat lymph), scald lymph and 4-PBA group (B, stimulated by 5% scalded rat lymph and 5mM 4-PBA) and control group(C,5% sham scalded rats lymph). Cells apoptosis was determined by flow cytometery with annexin V/PI. Expressions of protein of GRP78, CHOP and Caspase 12 were determined by western blotting.  Results   Significant apoptotic cells were determined in group A, which was higher than that of group C(P<0.01). There was significant decrease of apoptosis of cell treated with 4-PBA and burn lymph. Protein level of GRP78, CHOP and Caspase12 increased significantly when treated with burn lymph, but decreased when treated by 4-PBA(P<0.01). Conclusion   Lymph from rats with severe scalds can induce significant apoptosis in rPMECs. UPR may be the main pathway of apoptosis, which can be improved by 4-PBA.

Key words: Lymph; Endothelial cell; Unfolded protein response; Apoptosis; Lung injury

中图分类号: 

  • R644
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