中国循环杂志
中國循環雜誌
중국순배잡지
Chinese Circulation Journal
2015年
9期
879-883
,共5页
邓应忠%曹晨%郑兴萍%薛锐%刘芳%胡鄂曼%谭启荣
鄧應忠%曹晨%鄭興萍%薛銳%劉芳%鬍鄂曼%譚啟榮
산응충%조신%정흥평%설예%류방%호악만%담계영
脂联素%糖尿病%缺血再灌注损伤%缺血后处理%心肌保护
脂聯素%糖尿病%缺血再灌註損傷%缺血後處理%心肌保護
지련소%당뇨병%결혈재관주손상%결혈후처리%심기보호
Adiponectin%Diabetes mellitus%Ischemia-reperfusion injury%Ischemic post-conditioning%Myocardial protection
目的:通过建立大鼠糖尿病心肌缺血再灌注模型及缺血后处理模型,探讨脂联素在糖尿病心肌缺血再灌注损伤及缺血后处理中的变化及分子机制。方法:雄性SD大鼠80只,随机分为6组:正常假手术组(n=8)、正常心肌缺血再灌注组(n=16)、正常缺血后处理组(n=16)、糖尿病假手术组(n=8)、糖尿病心肌缺血再灌注组(n=16)和糖尿病缺血后处理组(n=16)。腹腔注射链脲佐菌素(STZ)的方法建立2型糖尿病模型,缺血再灌注模型采用结扎左冠状动脉前降支30 min再灌注120 min的方法建立;缺血后处理于再灌注前给予3个循环再灌注10 s,缺血10 s的处理;假手术模型仅以丝线穿过冠状动脉前降支但不结扎。再灌注120 min后处死大鼠,取心肌组织,三苯基氯化四氮唑法(TTC法)测定梗死面积;采用酶联免疫吸附试验(ELISA)检测血浆中脂联素的含量。免疫印迹方法(Western blot方法)测定心肌组织磷酸化蛋白激酶B (p-Akt)和总蛋白激酶B(Total-Akt)表达。结果:与正常心肌缺血再灌注组比较,正常缺血后处理组心肌梗死面积显著减小(P<0.05),而糖尿病心肌缺血再灌注组及糖尿病缺血后处理组的心肌梗死面积显著增加(P<0.01);与正常假手术组比较,正常心肌缺血再灌注组和正常缺血后处理组血清脂联素和p-Akt蛋白表达显著上调(P<0.05),糖尿病假手术组p-Akt蛋白表达显著下调(P<0.05);与正常缺血后处理组比较,糖尿病假手术组、糖尿病心肌缺血再灌注组及糖尿病缺血后处理组脂联素和p-Akt蛋白表达显著下调(P<0.05),差异均有统计学意义。线性相关分析显示,血浆脂联素的表达水平与心肌梗死面积呈负相关,而与心肌组织中p-Akt表达水平呈正相关,两者的相关系数分别为0.63和0.65(P<0.01)。结论:糖尿病血清脂联素表达降低,可致磷脂酰肌醇3-蛋白激酶B(PI3K/Akt)通路失活,导致糖尿病缺血再灌注损伤加重,缺血后处理不能实现对糖尿病心肌的保护作用。
目的:通過建立大鼠糖尿病心肌缺血再灌註模型及缺血後處理模型,探討脂聯素在糖尿病心肌缺血再灌註損傷及缺血後處理中的變化及分子機製。方法:雄性SD大鼠80隻,隨機分為6組:正常假手術組(n=8)、正常心肌缺血再灌註組(n=16)、正常缺血後處理組(n=16)、糖尿病假手術組(n=8)、糖尿病心肌缺血再灌註組(n=16)和糖尿病缺血後處理組(n=16)。腹腔註射鏈脲佐菌素(STZ)的方法建立2型糖尿病模型,缺血再灌註模型採用結扎左冠狀動脈前降支30 min再灌註120 min的方法建立;缺血後處理于再灌註前給予3箇循環再灌註10 s,缺血10 s的處理;假手術模型僅以絲線穿過冠狀動脈前降支但不結扎。再灌註120 min後處死大鼠,取心肌組織,三苯基氯化四氮唑法(TTC法)測定梗死麵積;採用酶聯免疫吸附試驗(ELISA)檢測血漿中脂聯素的含量。免疫印跡方法(Western blot方法)測定心肌組織燐痠化蛋白激酶B (p-Akt)和總蛋白激酶B(Total-Akt)錶達。結果:與正常心肌缺血再灌註組比較,正常缺血後處理組心肌梗死麵積顯著減小(P<0.05),而糖尿病心肌缺血再灌註組及糖尿病缺血後處理組的心肌梗死麵積顯著增加(P<0.01);與正常假手術組比較,正常心肌缺血再灌註組和正常缺血後處理組血清脂聯素和p-Akt蛋白錶達顯著上調(P<0.05),糖尿病假手術組p-Akt蛋白錶達顯著下調(P<0.05);與正常缺血後處理組比較,糖尿病假手術組、糖尿病心肌缺血再灌註組及糖尿病缺血後處理組脂聯素和p-Akt蛋白錶達顯著下調(P<0.05),差異均有統計學意義。線性相關分析顯示,血漿脂聯素的錶達水平與心肌梗死麵積呈負相關,而與心肌組織中p-Akt錶達水平呈正相關,兩者的相關繫數分彆為0.63和0.65(P<0.01)。結論:糖尿病血清脂聯素錶達降低,可緻燐脂酰肌醇3-蛋白激酶B(PI3K/Akt)通路失活,導緻糖尿病缺血再灌註損傷加重,缺血後處理不能實現對糖尿病心肌的保護作用。
목적:통과건립대서당뇨병심기결혈재관주모형급결혈후처리모형,탐토지련소재당뇨병심기결혈재관주손상급결혈후처리중적변화급분자궤제。방법:웅성SD대서80지,수궤분위6조:정상가수술조(n=8)、정상심기결혈재관주조(n=16)、정상결혈후처리조(n=16)、당뇨병가수술조(n=8)、당뇨병심기결혈재관주조(n=16)화당뇨병결혈후처리조(n=16)。복강주사련뇨좌균소(STZ)적방법건립2형당뇨병모형,결혈재관주모형채용결찰좌관상동맥전강지30 min재관주120 min적방법건립;결혈후처리우재관주전급여3개순배재관주10 s,결혈10 s적처리;가수술모형부이사선천과관상동맥전강지단불결찰。재관주120 min후처사대서,취심기조직,삼분기록화사담서법(TTC법)측정경사면적;채용매련면역흡부시험(ELISA)검측혈장중지련소적함량。면역인적방법(Western blot방법)측정심기조직린산화단백격매B (p-Akt)화총단백격매B(Total-Akt)표체。결과:여정상심기결혈재관주조비교,정상결혈후처리조심기경사면적현저감소(P<0.05),이당뇨병심기결혈재관주조급당뇨병결혈후처리조적심기경사면적현저증가(P<0.01);여정상가수술조비교,정상심기결혈재관주조화정상결혈후처리조혈청지련소화p-Akt단백표체현저상조(P<0.05),당뇨병가수술조p-Akt단백표체현저하조(P<0.05);여정상결혈후처리조비교,당뇨병가수술조、당뇨병심기결혈재관주조급당뇨병결혈후처리조지련소화p-Akt단백표체현저하조(P<0.05),차이균유통계학의의。선성상관분석현시,혈장지련소적표체수평여심기경사면적정부상관,이여심기조직중p-Akt표체수평정정상관,량자적상관계수분별위0.63화0.65(P<0.01)。결론:당뇨병혈청지련소표체강저,가치린지선기순3-단백격매B(PI3K/Akt)통로실활,도치당뇨병결혈재관주손상가중,결혈후처리불능실현대당뇨병심기적보호작용。
Objective: To investigate the effect of adiponectin levels with its related mechanism in diabetic myocardial ischemia-reperfusion injury and ischemia post-conditioning in experimental rats. Methods: A total of 80 male SD rats were randomly divided into 6 groups: Normal sham (NS) group,n=8, Normal ischemia-reperfusion injury (NIRI) group,n=16, Normal ischemia post-conditioning (NIPO) group,n=16 and Diabetic mellitus sham (DMS) group,n=8, Diabetic mellitus ischemia-reperfusion injury (DMIRI) group,n=16, Diabetic mellitus ischemic post-conditioning (DMIPO) group,n=16. DM rats model was established by intraperitoneal injection of streptozotocin; IR model was established by occlusion of left anterior descending (LAD) coronary artery for 30 min followed by reperfusion for 120min; IPO model was established by 3 cycles of ischemia for 10s and reperfusion for10s; the rats in Sham group received silk line wrapping of LAD without occlusion. The myocardial infarction (MI) area was measured by TTC staining, plasma adiponectin level was examined by ELISA, the protein expressions of p-Akt and total-Akt were detected by Western blot analysis. Results: Compared with NIRI group, NIPO group had decreased MI area,P<0.05, while DMIRI group and DMIPO group had increased MI area,P<0.01; compared with NS group, NIRI group and NIPO group showed up-regulated expression of adiponectin and p-Akt,P<0.05 and DMS group showed down-regulated p-Akt,P<0.05. Compared with NIPO group, three DM groups presented down-regulated adiponectin and p-Akt,P<0.05. Linear correlation analysis indicated that plasma adiponectin expression level was negatively related to MI area and positively related to myocardial tissue p-Akt expression with the correlation coefifcient at 0.63 and 0.65 respectively, P<0.01. Conclusion: Down-regulated plasma adiponectin expression may cause the inactivation of PI3K/Akt signal pathway and therefore aggravate DM ischemia-reperfusion injury which cannot be protected by ischemic post-conditioning in experimental rats.