目的 :研究重組質(zhì)粒p EGFP-N2/Pim-3轉(zhuǎn)染對(duì)內(nèi)毒素脂多糖（lipopolysaccharide,LPS）引起的大鼠胰腺AR4-2J細(xì)胞損傷的保護(hù)機(jī)制。方法:實(shí)驗(yàn)分為4組:A組（正常對(duì)照組）,B組（5μg/m L LPS處理組）,C組（空載質(zhì)粒p EGFP-N2轉(zhuǎn)染+5μg/m L LPS處理組）,D組（重組質(zhì)粒p EGFP-N2/Pim-3轉(zhuǎn)染+5μg/m L LPS處理組）。流式細(xì)胞儀檢測各組處理24 h后細(xì)胞凋亡情況。處理0、6、12、24 h后分別提取各組總RNA及蛋白,RT-PCR及Western blot分別檢測Pim-3、細(xì)胞間黏附分子（intercellular cell adhesion molecule-1,ICAM-1）、緊密連接蛋白閉鎖蛋白（Occludin）m RNA及蛋白的表達(dá)。結(jié)果 :處理24 h,各組細(xì)胞凋亡率分別為:A組（7.85±1.14）%、B組（53.13±5.73）%、C組（51.76±5.17）%、D組（21.13±4.15）%,D組細(xì)胞相對(duì)B、C組凋亡明顯減少,差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）;D組Pim-3表達(dá)與A、B、C組之間... 

【文章頁數(shù)】：6 頁

【文章目錄】：
1材料和方法
    1.1材料
    1.2方法
        1.2.1實(shí)驗(yàn)分組
        1.2.2細(xì)胞培養(yǎng)
        1.2.3質(zhì)粒提取及細(xì)胞轉(zhuǎn)染
        1.2.4 LPS處理細(xì)胞
        1.2.5流式細(xì)胞儀檢測AR4-2J細(xì)胞的凋亡情況
        1.2.6 RT-PCR檢測Pim-3、ICAM-1、Occludin m RNA表達(dá)
        1.2.7Western blot檢測Pim-3、ICAM-1、Occludin蛋白表達(dá)
    1.3統(tǒng)計(jì)學(xué)方法
2結(jié)果
    2.1質(zhì)粒轉(zhuǎn)染成功的鑒定
    2.2細(xì)胞凋亡檢測結(jié)果
    2.3 RT-PCR結(jié)果
    2.4 Western blot結(jié)果
3討論


【參考文獻(xiàn)】：
期刊論文
[1]Roles of BN52021 in platelet-activating factor pathway in inflammatory MS1 cells[J]. Shi-Hai Xia,Xiao-Hui Xiang,Kai Chen,Wei Xu.  World Journal of Gastroenterology. 2013(25)
[2]Serum inter-cellular adhesion molecule 1 is an early marker of diagnosis and prediction of severe acute pancreatitis[J]. Hai-Hang Zhu,Lin-Lin Jiang,Department of Gastroenterology,Northern Jiangsu Hospital,College of Clinical Medical Medicine,Yangzhou University,Yangzhou 225001,Jiangsu Province,China.  World Journal of Gastroenterology. 2012(20)
[3]Emodin promoted pancreatic claudin-5 and occludin expression in experimental acute pancreatitis rats[J]. Xian-Ming Xia,Shi-Mei Xing,Hai-Ling Ruan,Department of Gastroenterology and Hepatology,the Fourth Affiliated Hospital of Anhui Medical University,Hefei 230022,Anhui Province,China Bang-Ku Li,Department of Gastroenterology and Hepatology,the First Affiliated Hospital of Anhui Medical University,Hefei 230022,Anhui Province,China.  World Journal of Gastroenterology. 2012(17)



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