文章摘要
简 捷,黄 缘,刘利珍,李 双,邓 峰.重组质粒pEGFP-N2/Pim-3转染对胰腺细胞损伤的保护作用[J].南京医科大学学报,2016,(6):710~715
重组质粒pEGFP-N2/Pim-3转染对胰腺细胞损伤的保护作用
Protective role of recombinant plasmid pEGFP-N2/Pim-3 transfecting in rat pancreatic acinar cells damaged by lipopolysaccharide
投稿时间:2015-11-08  
DOI:10.7655/NYDXBNS20160613
中文关键词: 丝/苏氨酸激酶Pim-3  紧密连接蛋白闭锁蛋白  细胞间黏附分子  凋亡  脂多糖
英文关键词: Pim-3  Occuludin  ICAM-1  apoptosis  lipopolysaccharide
基金项目:
作者单位
简 捷 南昌大学第三附属医院消化内科,江西 南昌 330008 
黄 缘 南昌大学第二附属医院消化内科,江西 南昌 330006 
刘利珍 上海市嘉定区中心医院肿瘤科,上海 201800 
李 双 南昌大学第三附属医院消化内科,江西 南昌 330008 
邓 峰 南昌大学第三附属医院消化内科,江西 南昌 330008 
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中文摘要:
      目的:研究重组质粒pEGFP-N2/Pim-3转染对内毒素脂多糖(lipopolysaccharide,LPS)引起的大鼠胰腺AR4-2J细胞损伤的保护机制?方法:实验分为4组:A组(正常对照组),B组(5 μg/mL LPS处理组),C组(空载质粒pEGFP-N2转染+5 μg/mL LPS处理组),D组(重组质粒pEGFP-N2/Pim-3转染+5 ?滋g/mL LPS处理组)?流式细胞仪检测各组处理24 h后细胞凋亡情况?处理0?6?12?24 h后分别提取各组总RNA及蛋白,RT-PCR及Western blot分别检测Pim-3?细胞间黏附分子(intercellular cell adhesion molecule-1,ICAM-1)?紧密连接蛋白闭锁蛋白(Occludin) mRNA及蛋白的表达?结果:处理24 h,各组细胞凋亡率分别为:A组 (7.85 ± 1.14)%?B组 (53.13 ± 5.73)%? C组 (51.76 ± 5.17)%?D组 (21.13 ± 4.15)%,D组细胞相对B?C组凋亡明显减少,差异有统计学意义(P < 0.05);D组Pim-3表达与A?B?C组之间的差异有统计学意义(P < 0.05);处理12 h后 B?C?D组ICAM-1高表达,24 h达高峰,相对于A组,差异均有统计学意义(P < 0.01),D组与B?C组差异有统计学意义(P < 0.05);处理6 h后,B?C?D组Occludin高表达,12 h达高峰,相对于A组,差异均有统计学意义(P < 0.01),D组与B?C组差异有统计学意义(P < 0.05)?结论:丝/苏氨酸激酶Pim-3能够抑制胰腺细胞炎症反应和胰腺细胞凋亡,可能与上调闭锁蛋白和下调细胞间黏附分子的表达有关?
英文摘要:
      Objective:To explore the protective role of recombinant plasmid pEGFP-N2/Pim-3 transfecting in rat pancreatic acinar cells AR4-2J damaged by lipopolysaccharide (LPS). Methods: AR4-2J cells were divided into 4 groups: Group A, as the normal control group; Group B, treated with 5 μg/mL of LPS; Group C, transfected with blank plasmid pEGFP-N2 and treated with 5 ?滋g/mL of LPS; Group D, transfected with recombinant plasmid pEGFP-N2 /Pim-3 and treated with 5 μg/mL of LPS. Flow cytometry examined the apoptosis of AR4-2J cells in four groups after 24 h. RT-PCR was performed to detect the mRNA expression of Pim-3, intercellular cell adhesion molecule-1 (ICAM-1), and Occludin in AR4-2J cells of four groups treated in 0, 6, 12, 24 h, and Western blot was performed to detect the protein expression of Pim-3, ICAM-1, and Occludin in each group. Results: The apoptotic rates of Groups B and C were (53.13 ± 5.73)% and (51.76 ± 5.17)%, respectively, both significantly higher than that of Group D (21.13 ± 4.15)%(both P < 0.05). The expressions of Pim-3 mRNA and protein in Groups A, B and C remained a low level. The Pim-3 expression of Group D was significantly higher than those in Groups A, B and C (both P < 0.05). The ICAM-1 mRNA and protein expression levels of Groups B, C and D were constantly upregulated 6 h later, all significantly higher than those of Group A (all P < 0.01), and the ICAM-1 expressions of Groups B and C were significantly higher than Group D (both P < 0.05). The Occludin mRNA and protein expression levels of Groups B, C and D began to increase 6 h later, and peaked 12 h later, all significantly higher than those of Group A (all P < 0.01), and the Occludin expressions of Groups B and C were significantly lower than that of Group D (both P < 0.05). Conclusion: Pim-3 gene can inhibit the apoptosis of pancreatic acinar cell damaged by LPS, and also can upregulate the expression of Occludin and downregulate the expression of ICAM-1 to inhibit the pancreatic inflammatory reaction induced by LPS.
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