1-磷脂酰肌醇3-激酶%细胞外信号调节MAP激酶类%吗啡%缺血预处理%心力衰竭%心肌再灌注损伤
1-燐脂酰肌醇3-激酶%細胞外信號調節MAP激酶類%嗎啡%缺血預處理%心力衰竭%心肌再灌註損傷
1-린지선기순3-격매%세포외신호조절MAP격매류%마배%결혈예처리%심력쇠갈%심기재관주손상
1-phosphatidylinositol 3-kinase%Extracellular signal-regulated MAP kinases%Morphine%Ischemic preconditioning%Heart failure%Myocardial reperfusion injury
目的 评价1-磷脂酰肌醇3-激酶(PI3K)和细胞外信号调节激酶(ERK)信号通路在吗啡预处理减轻心力衰竭大鼠离体心脏缺血再灌注损伤中的作用.方法 雄性成年SD大鼠,体重200～230 g,尾静脉注射盐酸多柔比星2 mg/kg,1次/周,连续6周,制备大鼠慢性心力衰竭模型.第8周末将成功制备慢性心力衰竭模型的大鼠42只,采用随机数字表法分为7组(n=6):假手术组(S组)、缺血再灌注组(I/R组)、吗啡预处理组(MP组)、ERK抑制剂PD98059+吗啡预处理组(PD+ MP组)、PI3K抑制剂渥曼青霉素+吗啡预处理组(WT+ MP组)、PD98059组(PD组)和渥曼青霉素组(WT组).采用Langendorff离体心脏灌注及结扎左冠状动脉前降支(LAD)法制备大鼠缺血再灌注损伤模型,缺血30min,再灌注120 min.S组只缝线,不结扎LAD,持续灌注K-H液195 min.I/R组先灌注K-H液45 min,再制备大鼠缺血再灌注损伤模型.MP组先灌注K-H液15 min,再灌注含1μmol/L吗啡的K-H液进行预处理,灌注5 min,再次灌注K-H液5min,共3个循环,然后制备心脏缺血再灌注损伤模型.PD+ MP组和WT+ MP组于吗啡预处理前10 min,分别用含ERK抑制剂PD98059(10 μmol/L)和PI3K抑制剂渥曼青霉素(100 nmol/L)的K-H液进行灌注,持续至缺血5 min.PD组和WT组于缺血前40 min持续至缺血5 min分别灌注含PD98059和渥曼青霉素的K-H液.各组分别于心脏稳定灌注15 min、再灌注5和10 min时,收集冠脉流出液,采用化学比色法检测LDH活性.再灌注120 rin时,取下大鼠心脏,测量缺血危险区体积(AAR)、梗死区体积(IS)及IS/AAR比值.结果 与S组比较,I/R组再灌注5和10min时LDH活性升高,IS和IS/AAR均增加(P＜0.05),AAR差异无统计学意义(P＞0.05);与I/R组比较,MP组再灌注5min时LDH活性降低,IS和IS/AAR减小(P＜0.05),AAR差异无统计学意义,WT组和PD组LDH活性、IS、AAR和IS/AAR差异无统计学意义(P＞0.05);与MP组比较,PD+ MP组再灌注5和10 min时LDH活性升高,IS和IS/AAR减小(P＜0.05),WT+ MP组LDH活性、IS、AAR和IS/AAR差异无统计学意义(P＞0.05).结论 ERK信号通路的激活参与了吗啡预处理减轻心力衰竭大鼠离体心脏缺血再灌注损伤,而PI3K信号通路无此作用.
目的 評價1-燐脂酰肌醇3-激酶(PI3K)和細胞外信號調節激酶(ERK)信號通路在嗎啡預處理減輕心力衰竭大鼠離體心髒缺血再灌註損傷中的作用.方法 雄性成年SD大鼠,體重200～230 g,尾靜脈註射鹽痠多柔比星2 mg/kg,1次/週,連續6週,製備大鼠慢性心力衰竭模型.第8週末將成功製備慢性心力衰竭模型的大鼠42隻,採用隨機數字錶法分為7組(n=6):假手術組(S組)、缺血再灌註組(I/R組)、嗎啡預處理組(MP組)、ERK抑製劑PD98059+嗎啡預處理組(PD+ MP組)、PI3K抑製劑渥曼青黴素+嗎啡預處理組(WT+ MP組)、PD98059組(PD組)和渥曼青黴素組(WT組).採用Langendorff離體心髒灌註及結扎左冠狀動脈前降支(LAD)法製備大鼠缺血再灌註損傷模型,缺血30min,再灌註120 min.S組隻縫線,不結扎LAD,持續灌註K-H液195 min.I/R組先灌註K-H液45 min,再製備大鼠缺血再灌註損傷模型.MP組先灌註K-H液15 min,再灌註含1μmol/L嗎啡的K-H液進行預處理,灌註5 min,再次灌註K-H液5min,共3箇循環,然後製備心髒缺血再灌註損傷模型.PD+ MP組和WT+ MP組于嗎啡預處理前10 min,分彆用含ERK抑製劑PD98059(10 μmol/L)和PI3K抑製劑渥曼青黴素(100 nmol/L)的K-H液進行灌註,持續至缺血5 min.PD組和WT組于缺血前40 min持續至缺血5 min分彆灌註含PD98059和渥曼青黴素的K-H液.各組分彆于心髒穩定灌註15 min、再灌註5和10 min時,收集冠脈流齣液,採用化學比色法檢測LDH活性.再灌註120 rin時,取下大鼠心髒,測量缺血危險區體積(AAR)、梗死區體積(IS)及IS/AAR比值.結果 與S組比較,I/R組再灌註5和10min時LDH活性升高,IS和IS/AAR均增加(P＜0.05),AAR差異無統計學意義(P＞0.05);與I/R組比較,MP組再灌註5min時LDH活性降低,IS和IS/AAR減小(P＜0.05),AAR差異無統計學意義,WT組和PD組LDH活性、IS、AAR和IS/AAR差異無統計學意義(P＞0.05);與MP組比較,PD+ MP組再灌註5和10 min時LDH活性升高,IS和IS/AAR減小(P＜0.05),WT+ MP組LDH活性、IS、AAR和IS/AAR差異無統計學意義(P＞0.05).結論 ERK信號通路的激活參與瞭嗎啡預處理減輕心力衰竭大鼠離體心髒缺血再灌註損傷,而PI3K信號通路無此作用.
목적 평개1-린지선기순3-격매(PI3K)화세포외신호조절격매(ERK)신호통로재마배예처리감경심력쇠갈대서리체심장결혈재관주손상중적작용.방법 웅성성년SD대서,체중200～230 g,미정맥주사염산다유비성2 mg/kg,1차/주,련속6주,제비대서만성심력쇠갈모형.제8주말장성공제비만성심력쇠갈모형적대서42지,채용수궤수자표법분위7조(n=6):가수술조(S조)、결혈재관주조(I/R조)、마배예처리조(MP조)、ERK억제제PD98059+마배예처리조(PD+ MP조)、PI3K억제제악만청매소+마배예처리조(WT+ MP조)、PD98059조(PD조)화악만청매소조(WT조).채용Langendorff리체심장관주급결찰좌관상동맥전강지(LAD)법제비대서결혈재관주손상모형,결혈30min,재관주120 min.S조지봉선,불결찰LAD,지속관주K-H액195 min.I/R조선관주K-H액45 min,재제비대서결혈재관주손상모형.MP조선관주K-H액15 min,재관주함1μmol/L마배적K-H액진행예처리,관주5 min,재차관주K-H액5min,공3개순배,연후제비심장결혈재관주손상모형.PD+ MP조화WT+ MP조우마배예처리전10 min,분별용함ERK억제제PD98059(10 μmol/L)화PI3K억제제악만청매소(100 nmol/L)적K-H액진행관주,지속지결혈5 min.PD조화WT조우결혈전40 min지속지결혈5 min분별관주함PD98059화악만청매소적K-H액.각조분별우심장은정관주15 min、재관주5화10 min시,수집관맥류출액,채용화학비색법검측LDH활성.재관주120 rin시,취하대서심장,측량결혈위험구체적(AAR)、경사구체적(IS)급IS/AAR비치.결과 여S조비교,I/R조재관주5화10min시LDH활성승고,IS화IS/AAR균증가(P＜0.05),AAR차이무통계학의의(P＞0.05);여I/R조비교,MP조재관주5min시LDH활성강저,IS화IS/AAR감소(P＜0.05),AAR차이무통계학의의,WT조화PD조LDH활성、IS、AAR화IS/AAR차이무통계학의의(P＞0.05);여MP조비교,PD+ MP조재관주5화10 min시LDH활성승고,IS화IS/AAR감소(P＜0.05),WT+ MP조LDH활성、IS、AAR화IS/AAR차이무통계학의의(P＞0.05).결론 ERK신호통로적격활삼여료마배예처리감경심력쇠갈대서리체심장결혈재관주손상,이PI3K신호통로무차작용.
Objective To evaluate the roles of 1-phosphatidylinositol 3-kinase (PI3K) and extracellular signal-regulated kinase (ERK) signaling pathways in reduction of ischemia-reperfusion (I/R) injury to the isolated hearts by morphine preconditioning in the rats with chronic heart failure.Methods Adult male Sprague-Dawley rats,weighing 200-230 g,in which doxorubicin 2.0 mg/kg was injected via the tail vein once a week for 6 weeks to induce chronic heart failure,were studied.At the end of 8th week,42 rats with chronic heart failure were randomly divided into 7 groups (n =6 each) using a random number table:sham operation group (group S),I/R group,morphine preconditioning group (group MP),PD98059 (ERK inhibitor) + morphine preconditioning group (group PD + MP),wortmannin (PI3K inhibitor) + morphine preconditioning group (group WT + MP),PD98059 group (group PD) and wortmannin group (group WT).The hearts were quickly excised and passively perfused in a Langendorff apparatus and subjected to 30 min of occlusion of the left coronary artery followed by 2 h of reperfusion to establish the model of I/R injury.In group S,the hearts were only sutured,but not ligated and were continuously perfused with K-H solution for 195 min.In group I/R,the hearts were perfused with K-H solution for 45 min before ischemia.In group MP,the hearts were perfused with K-H solution for 15 min,with K-H solution containing morphine 1 μmol/L for 5 min and then with K-H solution for 5 min (3 cycles in total) before ischemia.In PD + MP and WT + MP groups,the hearts were perfused with K-H solution containing PD98059 (10 μmol/L) and wortmannin (100 nmol/L),respectively,starting from 10 min before morphine preconditioning until 5 min of ischemia.In PD and WT groups,the hearts were perfused with K-H solution containing PD98059 (10 μmol/L) and wortmannin (100 nmol/L),respectively,starting from 40 min before ischemia until 5 rin of ischemia.At 15 min of equilibration (baseline) and 5 and 10 min of reperfusion,the coronary flow was collected to detect the activity of lactate dehydrogenase (LDH).Infarct size (IS) and area at risk (AAR) were measured at the end of reperfusion and IS/AAR ratio was calculated.Results Compared with group S,LDH activity was significantly increased at 5 and 10 min of reperfusion,IS and IS/AAR ratio were also increased (P ＜ 0.05),and no significant change was found in AAR in group I/R (P ＞ 0.05).Compared with group I/R,LDH activity was significantly decreased at 5 min of reperfusion,IS and IS/AAR ratio were also decreased (P ＜ 0.05),and no significant change was found in AAR in group MP,and no significant change was found in LDH activity,IS,AAR and IS/AAR ratio in WT and PD groups (P ＞ 0.05).Compared with group MP,LDH activity was significantly increased at 5 and 10 min of reperfusion (P ＜ 0.05),and IS and IS/AAR ratio were decreased in group PD + MP,and no significant change was found in LDH activity,IS,AAR and IS/AAR ratio in group WT + MP (P ＞ 0.05).Conclusion Activation of ERK signaling pathway is involved in reduction of I/R injury to isolated hearts by morphine preconditioning in rats with chronic heart failure,however,PI3K signaling pathway has no such effect.
