雨蛙肽联合脂多糖诱导重度急性胰腺炎体外细胞模型钙通道的变化  

Calcium channel change in cerulean plus lipopolysaccharide stimulated pancreatic acinar cells

作　　者：唐曦平[1] 唐国都[1] 方春芸[1] 梁志海[1] 张露艺[1] 覃蒙斌[1] 

机构地区：[1]广西医科大学第一附属医院消化内科,广西南宁530021

出　　处：《中国现代医学杂志》2014年第18期7-11,共5页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:81060043);广西2011年研究生教育创新计划资助项目(No:2011105981002D31)

摘　　要：目的应用雨蛙肽联合脂多糖诱导AR42J细胞构建重度急性胰腺炎体外细胞模型,研究该模型构建前后细胞内钙通道的变化。方法体外培养大鼠AR42J细胞,实验分组设对照组、雨蛙肽处理组、雨蛙肽+脂多糖处理组。采用RT-PCR检测细胞Cav1.3、Cav2.1、Cav2.2、Cav3.1、Cav3.2 mRNA表达;Western blotting检测细胞Cav1.3、Cav2.1、Cav3.1蛋白表达水平;以荧光探针Fluo-4-AM标记细胞内游离钙,激光共聚焦扫描显微镜观察[Ca2+]i的变化。结果药物处理组Cav1.3、Cav2.1、Cav3.1 mRNA及蛋白表达水平较对照组均明显增高(mRNA:F=23.392,46.85,61.338;蛋白:F=33.798,43.588,79.467;P<0.01),雨蛙肽+脂多糖处理组与雨蛙肽处理组相比较上述检测指标的mRNA表达量也显著增高(P<0.01);药物处理组与对照组Cav2.2、Cav3.2mRNA表达水平无明显差异(F=0.175、0.316;P>0.05);激光共聚焦显微镜观察结果显示药物处理组比对照组[Ca2+]i升高(F=638.984,P<0.01),雨蛙肽+脂多糖处理组较雨蛙肽处理组[Ca2+]i也显著升高(P<0.01)。结论胰腺腺泡细胞内钙超载在雨蛙肽联合脂多糖诱导的重度AP细胞模型发生发展的机制中具有重要作用。【Objective】 To investigate the change of calcium channel in cells by applying cerulein plus lipopolysaccharide（LPS） to stimulate AR42J cells in order to build a cell model of severe acute pancreatitis.【Methods】Rat pancreatic acinar cell line AR42J were cultured in vitro. There were three groups： control group, cerulein treated group, cerulein plus LPS treated group. Expressions of Cav1.3, Cav 2.1, Cav 2.2, Cav 3.1, Cav 3.2 mRNA and were detected by RT-PCR. Protein expressions of Cav1.3, Cav 2.1, Cav 3.1 were detected byWestern blotting.Cells were loaded with Fluo-4-AM, and the change of [Ca^2＋]i in cells were determined by laser confocal scanning microscopy.【Results】Compared with the control, mRNA and protein expressions of Cav1.3, Cav 2.1 and Cav 3.1 in cells treated with drugs were increased（mRNA： F =23.392, 46.85, 61.338; protein： F =33.798, 43.588, 79.467; P〈0.01）. Compared with cerulein treated group, mRNA and protein expressions of Cav1.3, Cav 2.1 and Cav 3.1 in cells treated with cerulein plus LPS were increased significantly（P〈0.05）. There was no significant difference of Cav2.2and Cav3.2 mRNA expression in cells treated with or without drugs. Compared with the control, [Ca^2＋]i in cells treated with drugs was increased（F =638.984, P〈0.01）. And compared with the cerulein treated group, [Ca^2＋]i in cells treated with cerulein plus LPS was increased significantly（P〈0.01）.【Conclusions】Calcium overload in pancreatic acinar cells plays an important role in the mechanism of severe acute pancreatitis cell model with cerulein plus LPS stimulation.

关 键 词：重度急性胰腺炎 AR42J细胞 钙通道 雨蛙肽 脂多糖 

分 类 号：R576[医药卫生—消化系统]