姜黄素对肺缺血/再灌注损伤小鼠Caspase-12及细胞凋亡的影响  

Effect of Curcumin on Caspase-12 and Apoptosis in Pulmonary Ischemia/Reperfusion Injury Mice

作　　者：周俊辉[1,2] 赵珊[1] 陈海娥[1] 陈丹[1] 郝卯林[1] 应磊[1] 林丽娜[3] 王万铁[1] 

机构地区：[1]温州医科大学缺血/再灌注损伤研究所,浙江325035 [2]河南省胸科医院麻醉二科,郑州450008 [3]温州医科大学附属第一医院麻醉科,浙江325035

出　　处：《中国中西医结合杂志》2014年第9期1118-1124,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：浙江省公益技术应用研究项目(No.2011C37092);浙江省医药卫生科技计划项目(No.2010KYA132);浙江省中医药科技计划项目(No.2010ZB088);浙江省中医药重点学科建设计划(No.2012-XK-A28)

摘　　要：目的研究姜黄素(curcumin,CUR)对肺缺血/再灌注(ischemia/reperfusion,I/R)损伤小鼠天冬氨酸特异性半胱氨酸蛋白酶-12(cysteinyl aspartate specific proteinase-12,Caspase-12)及细胞凋亡的干预作用。方法采用C57BL/6J小鼠制作在体单侧肺原位I/R损伤模型。采用随机数字表法将60只小鼠分为6组,每组10只。假手术组(简称Sham组)、I/R组、I/R加二甲基亚砜(简称I/R加DMSO)组,I/R分别加100、150、200 mg/kg CUR(简称I/R加CUR-100、I/R加CUR-150、I/R加CUR-200)组。实验结束留取左肺,测定肺湿/干重比(W/D)和总肺水含量(TLW),光镜观察肺组织形态学改变,进行肺组织损伤定量评估(IQA),电镜观察肺组织超微结构改变,逆转录-聚合酶链反应(RT-PCR)和蛋白免疫印迹法(Western blot)分别检测Caspase-12和葡萄糖调节蛋白78(GRP78)mRNA及蛋白表达水平,原位末端标记(TUNEL)法检测肺组织细胞凋亡指数(AI)。结果与Sham组比较,I/R组Caspase-12和GRP78 mRNA及蛋白表达水平均升高(P<0.05),W/D、TLW、IQA和AI均增加(P<0.05,P<0.01),肺组织形态结构均发生明显损伤;与I/R加DMSO组比较,I/R加CUR-1 00组、I/R加CUR-1 50组、I/R加CUR-200组GRP78 mRNA及蛋白表达水平均升高(P<0.05),而Caspase-12 mRNA及蛋白表达水平均降低(P<0.05),W/D、TLW、IQA和AI亦均下降(P<0.05,P<0.01),肺组织形态学异常改变趋近于正常。结论姜黄素对I/R损伤发生的小鼠肺脏具有较好的保护作用,其机制可能与其对抗过度的未折叠蛋白反应(unfolded protein response,UPR)中Caspase-12引起的细胞凋亡有关。Objective To explore the effect of curcumin（CUR） on cycteinyl aspirate specific protease-12（Caspase-12） and pneumocyte apoptosis in pulmonary ischemia/reperfusion（l/R） injury mice.Methods The in vivo unilateral in situ pulmonary l/R injury mouse model was established in C57BL/6J mice.Sixty experimental mice were randomly divided into six groups by random digit table,i.e.,the sham-operation group（Sham）,the l/R group,the l/R＋dimethyl sulfoxide group（l/R ＋DMSO）,the l/R ＋ low dose CUR pre-treated group（l/R ＋ CUR-100）,the l/R ＋ middle dose CUR pre-treated group（I/R＋CUR-150）,the l/R ＋ high dose CUR pre-treated group（l/R ＋ CUR-200）,10 in each group.Mice were euthanized and their left lungs were excised.Wet lung weight to dry lung weight（W/D） and the total lung water content（TLW） were tested.The morphological changes of the lung tissue were observed and index of quantitative evaluation for alveolar damage（IQA） detected under light microscope.The ultra-microstructure of the lung tissue was observed under electron microscope.The mRNA and protein expression levels of Caspase-12 and glucose regulated protein（GRP78） were detected by reverse transcription-polymerase chain reaction（RT-PCR） and Western blot.Apoptosis index（Al） of the lung tissue was determined by terminal-deoxynucleoitidyl transferase mediated nick end labeling（TUNEL） method.Results Compared with the Sham group,expression levels of Caspase-12,GRP78 mRNA and protein all significantly increased in the l/R group（P〈0.05）;W/D,TLW,IQA,and Al were all notably higher（P〈0.05,P〈0.01）;the morphological and ultrastructural injury of the lung tissue were notably observed in I/R group.Compared with the l/R ＋DMSO group,expression levels of GRP78 mRNA and protein were increasingly higher in the l/R＋CUR-100 group,the l/R ＋ CUR-150 group,and the l/R＋CUR-200 group（P〈0.05）,expression levels of Caspase-12 mRNA and protein were lower（P〈0.05）;W/D,TLW,IQA,and Al also decreased�

关 键 词：CASPASE-12 肺 姜黄素 缺血/再灌注 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]