大黄素减轻胰腺腺泡AR42J细胞内质网应激研究.docVIP

大黄素减轻胰腺腺泡AR42J细胞内质网应激研究 　　[摘要]目的：探讨大黄素对胰腺腺泡AR42J细胞内质网应激的影响。方法：AR42J细胞以1×105 个/mL接种于6孔培养板中，分为正常对照组、模型组（雨蛙素终浓度为1×10lt;supgt;-7lt;/supgt; mol?Llt;supgt;-1lt;/supgt;， 脂多糖终浓度为10 mg?Llt;supgt;-1lt;/supgt;）、大黄素组（终浓度为10，20，40 μmol?Llt;supgt;-1lt;/supgt;），每组设3个复孔，培养24 h待细胞贴壁后弃上清，对照组加入单培、模型组加入用单培配置的造模液、治疗组在加入造模液前15～20 min分别加入不同浓度大黄素。37 ℃，5%COlt;subgt;2lt;/subgt;条件下继续培养3 h后，试剂盒检测细胞内蛋白酶、脂肪酶表达水平;光学显微镜观察细胞形态;激光共聚焦显微镜检测内质网钙离子水平;Western blot方法分析内质网应激相关信号分子的蛋白表达情况。结果：大黄素能显著抑制AR42J细胞合成蛋白酶、脂肪酶，减少细胞内钙离子水平，抑制内质网应激。结论：大黄素能够减轻雨蛙素联合脂多糖导致的胰腺腺泡细胞损伤，其作用机制与抑制细胞内钙超载及内质网应激有关。 　　[关键词]大黄素;AR42J细胞;内质网应激 　　[收稿日期]2014-05-15 　　[基金项目]国家自然科学基金项目;江苏高校优势学科建设工程项目（2011ZYX4-006） 　　[通信作者]*蔡宝昌，博士，教授，Tel：（025E-mail：bccai@126.comEffect of emodin in attenuating endoplasmic reticulum stress of 　　pancreatic acinar AR42J cellsWU Li， ZHANG Feng， ZHENG Shi-zhong， LU Yin， CAI Bao-chang* 　　（College of Pharmacy， Nanjing University of Chinese Medicine， Nanjing 210023， China）[Abstract]Objective： To explore the effect of emodin on endoplasmic reticulum （ER） stress of pancreatic acinar AR42J cells. Method： Rat pancreatic acinar AR42J cells were cultured in 6-well plates， and divided into the normal control group， the model group （with the final concentration at 1×10lt;supgt;-7lt;/supgt; mol?Llt;supgt;-1lt;/supgt; for cerulean and lipopolysaccharide at 10 mg?Llt;supgt;-1lt;/supgt;） and the emodin group （10， 20， 40 μmol?Llt;supgt;-1lt;/supgt;）. Cells in each group were cultured in three multiple pores for 24 h， and their supernate was removed after cell attachment. The normal control group was added with haploids， the model group was added with the modeling liquid for haploids， and the treatment groups were added with different concentrations of emodin at 15-20 min before the modeling liquid. The cells were continuously cultured for 3 h under 37 ℃ and 5%COlt;subgt;2lt;/subgt;. Their intracellular protease and lipase expressions were detected with kits. The cellular morphology was observed under opti