重组质粒pEGFP-N2/Pim-3转染对胰腺细胞损伤的保护作用

发布时间：2022-12-24 08:53

　　目的 :研究重组质粒p EGFP-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（5μg/m L LPS处理组）,C组（空载质粒p EGFP-N2转染+5μg/m L LPS处理组）,D组（重组质粒p EGFP-N2/Pim-3转染+5μg/m L 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）m RNA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... 

【文章页数】：6 页

【文章目录】：
1材料和方法
    1.1材料
    1.2方法
        1.2.1实验分组
        1.2.2细胞培养
        1.2.3质粒提取及细胞转染
        1.2.4 LPS处理细胞
        1.2.5流式细胞仪检测AR4-2J细胞的凋亡情况
        1.2.6 RT-PCR检测Pim-3、ICAM-1、Occludin m RNA表达
        1.2.7Western blot检测Pim-3、ICAM-1、Occludin蛋白表达
    1.3统计学方法
2结果
    2.1质粒转染成功的鉴定
    2.2细胞凋亡检测结果
    2.3 RT-PCR结果
    2.4 Western blot结果
3讨论


【参考文献】：
期刊论文
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