钙蛋白酶抑制剂对缺血-再灌注心肌肌浆网钙泵功能及其蛋白水平的影响  

Effects of calpain inhibitor on function and expression of sarcoplastic reticulum Ca^(2+)-ATPase in isolated ischemia-reperfusion hearts of rats

作　　者：王金桂[1] 祝宝华[2] 于晓頔[2] 陈开祥[2] 奚群英[2] 刘晨[2] 

机构地区：[1]无锡市中医医院ICU,214037 [2]东南大学附属扬州医院心血管病研究所

出　　处：《江苏医药》2011年第2期138-141,共4页Jiangsu Medical Journal

基　　金：江苏省135工程医学重点人才课题资助基金项目(RC2002016)

摘　　要：目的研究钙蛋白酶抑制剂ALLN对缺血-再灌注(I-R)大鼠心肌肌浆网钙泵(SERCA)功能的影响。方法 24只雄性SD大鼠随机分为正常灌注组(对照组)、I-R组、ALLN+I-R组和二甲亚砜+I-R组(DMSO+I-R组),分别检测再灌注15min时冠状动脉流出量(CF)和冠脉流出液中乳酸脱氢酶(LDH)漏出量;荧光分光光度计测定荧光强度来反映钙蛋白酶活性;分别采用改良p-NPP法和Millipore过滤技术测定SERCA的活性和ATP依赖的SERCA45Ca2+摄取率,免疫印迹法检测心肌细胞SERCA蛋白表达水平。结果与I-R组相比,ALLN+I-R组CF、SERCA蛋白表达水平、SERCA的活性和45Ca2+摄取率显著增高(P<0.01),而LDH漏出量和钙蛋白酶活性显著降低(P<0.01)。结论 ALLN对离体大鼠心脏I-R损伤有保护作用,与其抑制钙蛋白酶活性,一定程度减轻I-R损伤对SERCA功能及蛋白表达水平的影响有关。Objective To investigate the protective effects of calpain inhibitor（ALLN） on sarcoplastic reticulum Ca2＋-ATPase（SERCA） in isolated ischemia-reperfusion（I-R） hearts of rat.Methods Twenty-four male SD rats were randomly divided into 4 groups of control,I-R,ALLN＋I-R,and DMSO＋I-R with 6 rats each.At 15 min after reperfusion,coronary flux（CF） and LDH were examined,the activity of calpain was detected by fluorospectrophotometer,and SERCA activity and 45Ca2＋ uptake were evaluated with modified p-NPP and Millipore filting.The protein expression of SERCA was detected by immunoblotting.Results Compared with group I-R,CF,protein expression level and activity of SERCA and 45Ca2＋uptake in group ALLN＋I-R were all significantly increased（P0.01）,but LDH and the activity of calpain were significantly decreased（P0.01）.ConclusionALLN has a significant protective effect on I-R injury of isolated hearts,which could be associated with the depression of calpain activity and reduction of the effect of I-R injury on the protein expression of SERCA.

关 键 词：缺血-再灌注损伤 钙蛋白酶 肌浆网钙泵 

分 类 号：R541[医药卫生—心血管疾病]