中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2014年
3期
225-229
,共5页
白剑%顾蓉%王丙剑%张娜%康丽娜%徐标
白劍%顧蓉%王丙劍%張娜%康麗娜%徐標
백검%고용%왕병검%장나%강려나%서표
心力衰竭,充血性%细胞增殖%细胞凋亡%整合素连接激酶
心力衰竭,充血性%細胞增殖%細胞凋亡%整閤素連接激酶
심력쇠갈,충혈성%세포증식%세포조망%정합소련접격매
Heart failure,congestive%Cell proliferation%Apoptosis%Integrin-linked kinase
目的 观察心脏转染整合素连接激酶(ILK)对心力衰竭大鼠心功能的影响,并初步探讨其机制.方法 腹腔注射阿霉素(总剂量15 mg/kg,2周内分6次注射)建立大鼠心力衰竭模型.5周后通过心脏超声筛查,将成功建模的大鼠按照随机数字表法随机分为转染Ad-ILK组(Ad-ILK组)和转染Ad-null组(对照组),每组20只.通过开胸左心室多点注射Ad-ILK或Ad-null,4周后采用蛋白质免疫印迹法检测ILK蛋白表达水平及其活性,超声心动图、心导管法检测大鼠心功能,ELISA法检测大鼠血清B型利钠肽(BNP)表达水平,并采用TUNEL法检测心肌细胞凋亡率,磷酸化组蛋白H3免疫荧光染色检测心肌细胞增殖率.结果 转染4周后,Ad-ILK组大鼠心脏组织中ILK蛋白表达水平明显高于对照组(P＜0.05),其下游蛋白激酶B(Akt)磷酸化水平也明显高于对照组(P＜0.05).转染4周后,Ad-ILK组大鼠左心室射血分数和短轴缩短率均明显高于对照组[(60.56±2.61)％比(51.94±2.28)％和(28.10±1.83)％比(22.82±1.68)％,P均＜0.05],左心室舒张末期内径和收缩末期内径均明显小于对照组[(6.22±0.24) mm比(7.15±0.21) mm和(4.42±0.23) mm比(5.65±0.25) mm,P均＜0.05],左心室舒张末压明显低于对照组[(12.96±2.10) mmHg比(21.45±2.48) mmHg(1 mmHg =0.133 kPa),P＜0.05].Ad-ILK组大鼠血清B型利钠肽水平亦明显低于对照组(P＜0.05).Ad-ILK组心肌细胞凋亡率明显低于对照组[(0.23±0.02)％比(0.45±0.04)％,P＜0.05],而心肌细胞增殖率则显著高于对照组[(0.60±0.07)％比(0.24±0.03)％,P＜0.01].结论 心肌注射转染Ad-ILK能够明显改善心力衰竭大鼠心功能,这一作用可能与过表达ILK能够抑制心肌细胞凋亡、促进心肌细胞增殖有关.
目的 觀察心髒轉染整閤素連接激酶(ILK)對心力衰竭大鼠心功能的影響,併初步探討其機製.方法 腹腔註射阿黴素(總劑量15 mg/kg,2週內分6次註射)建立大鼠心力衰竭模型.5週後通過心髒超聲篩查,將成功建模的大鼠按照隨機數字錶法隨機分為轉染Ad-ILK組(Ad-ILK組)和轉染Ad-null組(對照組),每組20隻.通過開胸左心室多點註射Ad-ILK或Ad-null,4週後採用蛋白質免疫印跡法檢測ILK蛋白錶達水平及其活性,超聲心動圖、心導管法檢測大鼠心功能,ELISA法檢測大鼠血清B型利鈉肽(BNP)錶達水平,併採用TUNEL法檢測心肌細胞凋亡率,燐痠化組蛋白H3免疫熒光染色檢測心肌細胞增殖率.結果 轉染4週後,Ad-ILK組大鼠心髒組織中ILK蛋白錶達水平明顯高于對照組(P＜0.05),其下遊蛋白激酶B(Akt)燐痠化水平也明顯高于對照組(P＜0.05).轉染4週後,Ad-ILK組大鼠左心室射血分數和短軸縮短率均明顯高于對照組[(60.56±2.61)％比(51.94±2.28)％和(28.10±1.83)％比(22.82±1.68)％,P均＜0.05],左心室舒張末期內徑和收縮末期內徑均明顯小于對照組[(6.22±0.24) mm比(7.15±0.21) mm和(4.42±0.23) mm比(5.65±0.25) mm,P均＜0.05],左心室舒張末壓明顯低于對照組[(12.96±2.10) mmHg比(21.45±2.48) mmHg(1 mmHg =0.133 kPa),P＜0.05].Ad-ILK組大鼠血清B型利鈉肽水平亦明顯低于對照組(P＜0.05).Ad-ILK組心肌細胞凋亡率明顯低于對照組[(0.23±0.02)％比(0.45±0.04)％,P＜0.05],而心肌細胞增殖率則顯著高于對照組[(0.60±0.07)％比(0.24±0.03)％,P＜0.01].結論 心肌註射轉染Ad-ILK能夠明顯改善心力衰竭大鼠心功能,這一作用可能與過錶達ILK能夠抑製心肌細胞凋亡、促進心肌細胞增殖有關.
목적 관찰심장전염정합소련접격매(ILK)대심력쇠갈대서심공능적영향,병초보탐토기궤제.방법 복강주사아매소(총제량15 mg/kg,2주내분6차주사)건립대서심력쇠갈모형.5주후통과심장초성사사,장성공건모적대서안조수궤수자표법수궤분위전염Ad-ILK조(Ad-ILK조)화전염Ad-null조(대조조),매조20지.통과개흉좌심실다점주사Ad-ILK혹Ad-null,4주후채용단백질면역인적법검측ILK단백표체수평급기활성,초성심동도、심도관법검측대서심공능,ELISA법검측대서혈청B형리납태(BNP)표체수평,병채용TUNEL법검측심기세포조망솔,린산화조단백H3면역형광염색검측심기세포증식솔.결과 전염4주후,Ad-ILK조대서심장조직중ILK단백표체수평명현고우대조조(P＜0.05),기하유단백격매B(Akt)린산화수평야명현고우대조조(P＜0.05).전염4주후,Ad-ILK조대서좌심실사혈분수화단축축단솔균명현고우대조조[(60.56±2.61)％비(51.94±2.28)％화(28.10±1.83)％비(22.82±1.68)％,P균＜0.05],좌심실서장말기내경화수축말기내경균명현소우대조조[(6.22±0.24) mm비(7.15±0.21) mm화(4.42±0.23) mm비(5.65±0.25) mm,P균＜0.05],좌심실서장말압명현저우대조조[(12.96±2.10) mmHg비(21.45±2.48) mmHg(1 mmHg =0.133 kPa),P＜0.05].Ad-ILK조대서혈청B형리납태수평역명현저우대조조(P＜0.05).Ad-ILK조심기세포조망솔명현저우대조조[(0.23±0.02)％비(0.45±0.04)％,P＜0.05],이심기세포증식솔칙현저고우대조조[(0.60±0.07)％비(0.24±0.03)％,P＜0.01].결론 심기주사전염Ad-ILK능구명현개선심력쇠갈대서심공능,저일작용가능여과표체ILK능구억제심기세포조망、촉진심기세포증식유관.
Objective The aim of this study is to investigate the effects of cardiac integrin-linked kinase (ILK) overexpression in a rat model of doxorubicin-induced heart failure and the underlying mechanisms.Methods Rat heart failure model was induced by intraperitoneal administration of doxorubicin (6 injections within 2 weeks: total dose =15 mg/kg).Five weeks after the first injection,rats with heart failure were confirmed by echocardiography and then randomly divided into Ad-ILK group (intramyocardially injected with adenoviral vector expressing ILK) and Ad-null group (intramyocardially injected with empty ad-null,n =20 each).After 4 weeks,ILK expression and activity were detected by Western blot,cardiac function was determined by echocardiographic and hemodynamic examinations.Apoptosis was measured by TUNEL analysis and cardiomyocyte proliferation was estimated by phosphohistone-H3 staining.Results Western blot analysis revealed higher expression of ILK as well as the phosphorylation levels of Akt in AdILK hearts as compared with ad-null controls.Four weeks after transfection,LVEF and LVFS were significantly higher in Ad-ILK group as compared with control group [LVEF: (60.56 ± 2.61)％ vs.(51.94±2.28)％,P＜0.05; LVFS: (28.10± 1.83)％ vs.(22.82 ± 1.68)％,P ＜0.05].The LVEDD and LVESD,as well as LVEDP were significantly lower in Ad-ILK group compared with control group [LVEDD: (6.22 ± 0.24) mm vs.(7.15 ± 0.21) mm,P ＜ 0.05 ; LVESD: (4.42 ± 0.23) mm vs.(5.65±0.25) mm,P＜0.05; LVEDP: (12.96±2.10) mmHgvs.(21.45±2.48) mmHg(1 mmHg=0.133 kPa),P ＜0.05].Reduced levels of serum BNP was also seen in the Ad-ILK group.TUNEL analysis showed that ILK treatment significantly inhibited the apoptosis of cardiomyocytes [(0.23 ± 0.02) ％ vs.(0.45 ± 0.04)％,P ＜ 0.05].Moreover,increased cardiomyocyte proliferation was found in Ad-ILK group through the phospho-histone H3 staining [(0.60 ± 0.07) ％ vs.(0.24 ± 0.03) ％,P ＜ 0.01].Conclusion ILK gene therapy improves cardiac function in this rat model of heart failure,and is associated with reduced apoptosis and increased cardiomyocyte proliferation.
