腺相关病毒介导心肌肌浆网三磷酸腺苷酶基因治疗心力衰竭犬的安全性分析

目的研究心肌注射重组腺相关病毒(rAAV)_(2/1)介导心肌肌浆网Ca~(2+)-ATP酶(SERCA2a)基因治疗慢性心力衰竭犬的安全性。方法选取成年比格犬8只,随机分为对照组和SERCA2a基因转导组(转导组),每组4只。分别采用PCR、RT PCR和Western blot法从DNA、RNA和蛋白水平检测rAAV_(2/1)SERCA2a转导后在犬体内的分布;采用双夹心ELISA法检测转导组犬的血清。结果除注射rAAV_(2/1)的心肌区域外,未注射rAAV_(2/1)的心肌区域、主要脏器、骨骼肌和性腺中均未检测到rAAV_(2/1)-SERCA2a。对照组与转导组犬心肌、腰大肌、咀嚼肌和睾丸表现出相同的SERCA2a蛋白表达谱,未在其他组织中检测到。转导组犬血清内未检测到抗rAAV_(2/1)抗体。结论 rAAV_(2/1)介导的基因转导是相对安全的,未整合到心脏以外的其他组织,也未在犬体内引发体液免疫。     

姜黄素对心力衰竭兔肌浆网钙泵表达的影响

目的 探讨姜黄素对心力衰竭(心衰)兔肌浆网钙泵表达的影响.方法 采用主动脉瓣反流联合腹主动脉缩窄制作慢性心衰家兔模型.随机分为心衰姜黄素组、心衰安慰剂组、对照姜黄素组、对照安慰剂组.8周后计算心脏重量与体重比值,观察超微结构,检测肌浆网钙泵mRNA和蛋白的表达水平及活性.结果 心衰姜黄素组和心衰安慰剂组心脏重量与体重比值均大于对照组(P＜0.05);且心衰姜黄素组比值小于心衰安慰剂组(P＜0.05).电子显微镜显示心衰姜黄素组的心脏超微结构有所改善.心衰姜黄素组和心衰安慰剂组肌浆网钙泵mRNA、蛋白表达及活性均小于对照组(P＜0.05),但心衰姜黄素组均显著高于心衰安慰剂组(P＜0.05).结论 姜黄素能在mRNA水平和蛋白水平提高心衰家兔肌浆网钙泵的表达,提高肌浆网钙泵的活性,这可能是姜黄素改善心衰的机制之一.     

过表达肌浆网钙ATP酶治疗实验性慢性缺血性心力衰竭

目的 研究肌浆网钙ATP酶2a(SERCA2a)基因过表达对慢性缺血性心力衰竭(心衰)心功能及心肌内质网应激(ERS)相关凋亡的影响.方法采用ameroid环束扎小型猪前降支制备慢性缺血性心衰模型.开胸心肌内注射重组腺相关病毒以过表达SERCA2a或对照报告基因绿色荧光蛋白.60 d后检测血流动力学、SERCA2a的表达和活性、心肌凋亡及ERS标志蛋白-分子伴侣GRp78、凋亡蛋白caspase-12的表达.结果基因转导后60 d,与心衰对照及报告基因组相比,转基因组SERCA2a蛋白表达和活性显著增高,心功能参数改善,心肌凋亡指数降低,伴GRP78和活化caspase12表达下降.结论过表达SERCA2a可改善慢性缺血性心衰的心脏功能,其机制可能涉及减轻ERS相关的心肌细胞凋亡.

Abstract：

Objective Chronic myocardial ischemia (CMI) has become the most importat cause of heart failure (HF) all over the world. The aim of the current study was to investigate the effects of Sarcoendoplasmic reticulum calcium ATPaee 2a (SERCA2a) gene transfer on cardiac function and endoplasmic reticulum stress (ERS) associated myocardial apoptosis in a minipig HF animal model induced by CMI. Methods HF was induced in minipigs by implantation of ameroid constrictor in the initial segment of left anterior descending (LAD) branch of coronary artery. After confirmation of myocardial perfusion defects and cardiac function impairment by myocardial perfusion imaging and echocardiography, animals were divided into 4 groups (n =4 each): HF group, HF + enhanced green fluorescent protein (EGFP) group,HF + SERCA2a group, and shamed animals as control group. A total amount of 1×1012 v.g. Of rAAV1EGFP or rAAV1-SERCA2a were injected intramyocardially to each animal of HF + EGFP and HF +SERCA2a groups. Sixty days after gene transfer, protein level and activity of SERCA2a were examined,cardiac functions and changes of serum inflammatory and neuro-hormonal factors were determined. Apoptotic index of the ischemic myocardium, protein levels of ER stress marker glucose regulated protein 78 ( GRP 78) and ER stress specific apoptotic marker caspase-12 were also assayed. Results At the study end,echocardiographic and hemodynamic measurements indicated a significant improvement of both cardiac systolic and diastolic function in HF + SERCA2a group compared with HF/HF + EGFP groups [LVEF (60.2±8.6)%vs (44.2±7.1)% and (46.8±6.7)%, Ev/Ay 1.28±0.24 vs 0.77 ±0.17 and 0.80±0.21, +dp/dtmax(2713.9 ±434.0) mm Hg/s ( 1 mm Hg =0.133 kPa) vs (1892.3 ±434.2) mm Hg/s and (1931.2±397.4)mm Hg/s, -dp/dtmax (1422.1±334.4) mm Hg/s vs (848.3±308.3) mm Hg/s and (849.5±278.3)mm Hg/s, P＜0.05], along with increase in both SERCA2a protein level (1.13±0.26 vs 0.73 ±0.17 and 0.64±0.18, P＜0.05) and activity [(16.2±5.5) IU/ml vs (7.9±3.1) IU/ml and (7.5 ±2.8)IU/ml, P ＜0.05] compared with HF/HF + EGFP groups. Serum concentrations of inflammatory factor tumor necrotic factor α [(382.3±114.4) ng/L vs (732.3±201.4) ng/L and (689.8±192 5) ng/L, P＜0. 05], neural-hormonal factors brain natriuretic peptide [(142.6±45.3) ng/L vs (422.3±113.6) ng/L and(393.7 ±103.3)ng/L, P＜0.01], endothelin-1 [(111.4 ±37.5)ng/L vs (193.5 ±54.3)ng/L and (201.0±72.1)ng/L,P＜0.05] and angiotensin Ⅱ[(189.7±65.2)μg/L vs (538.3 ± 135.2) μg/L and ( 525.5±144.1)μg/L, P＜0.01] were also significantly decreased in HF + SERCA2a group compared with HF/HF + EGFP groups. The apoptotic index [(12.71±4.11)% vs(23.22±7.23) % and (24.31±6.38)%, P＜0.05], protein levels of GRP78 (1.27±0.33 vs 3.23±1.14 and 4.18±1.13, P＜0.05)and protein level ratios of cleaved caspase-12 to total caspase-12[(4.62±1.93)% vs (9.71±2.70)% and (10.14±2.81)%, P＜0.05] were also significantly reduced in the ischemic myocardium of HF+SERCA2a group compared with the HF/HF + EGFP groups. Conclusion Overexpression of SERCA2a significantly improved cardiac systolic and diastolic function in this HF model partly through attenuation of ER stress related myocardial apoptosis, suggesting its therapeutic potential for CM1 related heart failure.     

过表达肌浆网钙ATP酶治疗实验性慢性缺血性心力衰竭

Objective Chronic myocardial ischemia (CMI) has become the most importat cause of heart failure (HF) all over the world. The aim of the current study was to investigate the effects of Sarcoendoplasmic reticulum calcium ATPaee 2a (SERCA2a) gene transfer on cardiac function and endoplasmic reticulum stress (ERS) associated myocardial apoptosis in a minipig HF animal model induced by CMI. Methods HF was induced in minipigs by implantation of ameroid constrictor in the initial segment of left anterior descending (LAD) branch of coronary artery. After confirmation of myocardial perfusion defects and cardiac function impairment by myocardial perfusion imaging and echocardiography, animals were divided into 4 groups (n =4 each): HF group, HF + enhanced green fluorescent protein (EGFP) group,HF + SERCA2a group, and shamed animals as control group. A total amount of 1×1012 v.g. Of rAAV1EGFP or rAAV1-SERCA2a were injected intramyocardially to each animal of HF + EGFP and HF +SERCA2a groups. Sixty days after gene transfer, protein level and activity of SERCA2a were examined,cardiac functions and changes of serum inflammatory and neuro-hormonal factors were determined. Apoptotic index of the ischemic myocardium, protein levels of ER stress marker glucose regulated protein 78 ( GRP 78) and ER stress specific apoptotic marker caspase-12 were also assayed. Results At the study end,echocardiographic and hemodynamic measurements indicated a significant improvement of both cardiac systolic and diastolic function in HF + SERCA2a group compared with HF/HF + EGFP groups [LVEF (60.2±8.6)%vs (44.2±7.1)% and (46.8±6.7)%, Ev/Ay 1.28±0.24 vs 0.77 ±0.17 and 0.80±0.21, +dp/dtmax(2713.9 ±434.0) mm Hg/s ( 1 mm Hg =0.133 kPa) vs (1892.3 ±434.2) mm Hg/s and (1931.2±397.4)mm Hg/s, -dp/dtmax (1422.1±334.4) mm Hg/s vs (848.3±308.3) mm Hg/s and (849.5±278.3)mm Hg/s, P＜0.05], along with increase in both SERCA2a protein level (1.13±0.26 vs 0.73 ±0.17 and 0.64±0.18, P＜0.05) and activity [(16.2±5.5) IU/ml vs (7.9±3.1) IU/ml and (7.5 ±2.8)IU/ml, P ＜0.05] compared with HF/HF + EGFP groups. Serum concentrations of inflammatory factor tumor necrotic factor α [(382.3±114.4) ng/L vs (732.3±201.4) ng/L and (689.8±192 5) ng/L, P＜0. 05], neural-hormonal factors brain natriuretic peptide [(142.6±45.3) ng/L vs (422.3±113.6) ng/L and(393.7 ±103.3)ng/L, P＜0.01], endothelin-1 [(111.4 ±37.5)ng/L vs (193.5 ±54.3)ng/L and (201.0±72.1)ng/L,P＜0.05] and angiotensin Ⅱ[(189.7±65.2)μg/L vs (538.3 ± 135.2) μg/L and ( 525.5±144.1)μg/L, P＜0.01] were also significantly decreased in HF + SERCA2a group compared with HF/HF + EGFP groups. The apoptotic index [(12.71±4.11)% vs(23.22±7.23) % and (24.31±6.38)%, P＜0.05], protein levels of GRP78 (1.27±0.33 vs 3.23±1.14 and 4.18±1.13, P＜0.05)and protein level ratios of cleaved caspase-12 to total caspase-12[(4.62±1.93)% vs (9.71±2.70)% and (10.14±2.81)%, P＜0.05] were also significantly reduced in the ischemic myocardium of HF+SERCA2a group compared with the HF/HF + EGFP groups. Conclusion Overexpression of SERCA2a significantly improved cardiac systolic and diastolic function in this HF model partly through attenuation of ER stress related myocardial apoptosis, suggesting its therapeutic potential for CM1 related heart failure.     

过表达肌浆网钙ATP酶治疗实验性慢性缺血性心力衰竭

Objective Chronic myocardial ischemia (CMI) has become the most importat cause of heart failure (HF) all over the world. The aim of the current study was to investigate the effects of Sarcoendoplasmic reticulum calcium ATPaee 2a (SERCA2a) gene transfer on cardiac function and endoplasmic reticulum stress (ERS) associated myocardial apoptosis in a minipig HF animal model induced by CMI. Methods HF was induced in minipigs by implantation of ameroid constrictor in the initial segment of left anterior descending (LAD) branch of coronary artery. After confirmation of myocardial perfusion defects and cardiac function impairment by myocardial perfusion imaging and echocardiography, animals were divided into 4 groups (n =4 each): HF group, HF + enhanced green fluorescent protein (EGFP) group,HF + SERCA2a group, and shamed animals as control group. A total amount of 1×1012 v.g. Of rAAV1EGFP or rAAV1-SERCA2a were injected intramyocardially to each animal of HF + EGFP and HF +SERCA2a groups. Sixty days after gene transfer, protein level and activity of SERCA2a were examined,cardiac functions and changes of serum inflammatory and neuro-hormonal factors were determined. Apoptotic index of the ischemic myocardium, protein levels of ER stress marker glucose regulated protein 78 ( GRP 78) and ER stress specific apoptotic marker caspase-12 were also assayed. Results At the study end,echocardiographic and hemodynamic measurements indicated a significant improvement of both cardiac systolic and diastolic function in HF + SERCA2a group compared with HF/HF + EGFP groups [LVEF (60.2±8.6)%vs (44.2±7.1)% and (46.8±6.7)%, Ev/Ay 1.28±0.24 vs 0.77 ±0.17 and 0.80±0.21, +dp/dtmax(2713.9 ±434.0) mm Hg/s ( 1 mm Hg =0.133 kPa) vs (1892.3 ±434.2) mm Hg/s and (1931.2±397.4)mm Hg/s, -dp/dtmax (1422.1±334.4) mm Hg/s vs (848.3±308.3) mm Hg/s and (849.5±278.3)mm Hg/s, P＜0.05], along with increase in both SERCA2a protein level (1.13±0.26 vs 0.73 ±0.17 and 0.64±0.18, P＜0.05) and activity [(16.2±5.5) IU/ml vs (7.9±3.1) IU/ml and (7.5 ±2.8)IU/ml, P ＜0.05] compared with HF/HF + EGFP groups. Serum concentrations of inflammatory factor tumor necrotic factor α [(382.3±114.4) ng/L vs (732.3±201.4) ng/L and (689.8±192 5) ng/L, P＜0. 05], neural-hormonal factors brain natriuretic peptide [(142.6±45.3) ng/L vs (422.3±113.6) ng/L and(393.7 ±103.3)ng/L, P＜0.01], endothelin-1 [(111.4 ±37.5)ng/L vs (193.5 ±54.3)ng/L and (201.0±72.1)ng/L,P＜0.05] and angiotensin Ⅱ[(189.7±65.2)μg/L vs (538.3 ± 135.2) μg/L and ( 525.5±144.1)μg/L, P＜0.01] were also significantly decreased in HF + SERCA2a group compared with HF/HF + EGFP groups. The apoptotic index [(12.71±4.11)% vs(23.22±7.23) % and (24.31±6.38)%, P＜0.05], protein levels of GRP78 (1.27±0.33 vs 3.23±1.14 and 4.18±1.13, P＜0.05)and protein level ratios of cleaved caspase-12 to total caspase-12[(4.62±1.93)% vs (9.71±2.70)% and (10.14±2.81)%, P＜0.05] were also significantly reduced in the ischemic myocardium of HF+SERCA2a group compared with the HF/HF + EGFP groups. Conclusion Overexpression of SERCA2a significantly improved cardiac systolic and diastolic function in this HF model partly through attenuation of ER stress related myocardial apoptosis, suggesting its therapeutic potential for CM1 related heart failure.     

左心室射血分数保留的心力衰竭研究进展

<正>心力衰竭(心衰)是由于任何心脏结构或功能异常导致心室充盈或射血能力受损的一组复杂临床综合征,其主要临床表现为呼吸困难和乏力(活动耐量受限)以及液体潴留(肺淤血和外周水肿)。1定义及诊断标准在美国心脏病学学院基金会(American College of Cardiology Foundation,ACCF)和AHA联合发布的2013年心     

射血分数保留的心力衰竭研究进展

射血分数保留的心力衰竭拥有较为独特的发病因素、病理生理变化及预后,缺乏有效的诊断治疗方法.本文总结近年相关领域的研究结果,从流行病学、病因、病理生理、诊断及治疗等方面讨论射血分数保留的心力衰竭目前的研究进展.     

左室射血分数保留的心力衰竭

收缩性心力衰竭（systolic heart failure）指由于心脏泵血不能满足代谢需要所致心功能异常的一种病理生理状态。舒张性心力衰竭（diastolic heart faillife）指一组具有心力衰竭症状和（或）体征,以心室射血分数正常而舒张功能异常为特征的临床综合征。     

射血分数保留的心力衰竭的研究进展

<正>射血分数保留的心力衰竭(HFpEF)占心力衰竭总数的50%^[1]。来自社区动脉粥样硬化风险研究的数据表明,美国每年新诊断850 000例心力衰竭患者,其中一半为HFpEF。China-HF研究纳入国内132家医院13 687例心力衰竭患者数据分析显示,LVEF≥50%心力衰竭患者占心力衰竭总数的46.9%。随着我国人口老龄化进程加剧以及高血压、糖尿病、肥胖等疾病患病率不断增加,HFpEF患病率正在急剧增长。     

慢性心力衰竭基因治疗的临床研究进展

<正>慢性心力衰竭是大多数心血管病的最终阶段,是世界范围发病率和死亡率最主要的病因之一。据估计,目前全世界超过2300万人患有慢性心力衰竭,随着老龄化社会的到来,其发病率将会在未来10年继续增长。近几十年新的医学观念和手段已经使多种心脏疾病的预后获得改观,但慢性心力衰竭死亡率(5年死亡率近50%)依然高居不下~([1])。心脏移植是目前晚期心力衰竭患者最为有效的治疗手段,但供者不足、术后免疫抑制以及花费巨大严重阻碍其发展和推广。最     

肌浆网Ca2+ATP酶在充血性心力衰竭中的研究

充血性心力衰竭(CHF)是各种心脏病终末期的共同归宿,发病率和死亡率均较高.研究表明,肌浆网膜上几个关键调节分子的功能障碍引起心肌细胞内钙调节的内在缺损,在CHF的发病过程中发挥了重要的作用.本文对肌浆网Ca2 ATP酶的蛋白结构、调节机制,在CHF的变化及其在CHF基因治疗方面应用的进展作一综述.     

肌浆网钙ATP酶2a基因转导治疗心力衰竭的研究

心血管疾病和其他疾病造成的心力衰竭在世界各国影响数千万人。尽管最近内外科治疗有较大的进展,但病情较重患的5年生存率仍低于50％。转基因治疗作为防治心力衰竭的新思路之一,目前集中于3方面的研究：(1)促进血管再生,改善心肌缺血,预防心力衰竭,其中与血管再生有关的生长因子VEGF和bFGF已应用于急性心肌梗死临床试验。(2)抑制对移植心脏的免疫排斥反应。(3)增强心肌收缩力治疗晚期心力衰竭。旨在增强心肌收缩力的靶基因中包括肌浆网钙调节基因。现介绍有关此基因在治疗心力衰竭中的研究进展。     

射血分数保留性心力衰竭的研究进展

左心室射血分数保留性心力衰竭(HFpEF)旧称舒张功能不全的心力衰竭或收缩功能正常的心力衰竭,是一种日益流行的健康问题,相对于LVEF降低的心力衰竭(HFrEF)而言,其特征是LVEF正常或者接近正常,但有心力衰竭的临床表现。临床上该型患者的发病率逐渐增长,且预后比HFrEF更差。本文旨在对HFpEF目前的研究进展进行综诉。     

射血分数保留性心力衰竭动物模型的研究进展

近年来,射血分数保留性心力衰竭(HFpEF)成为严重威胁人类健康的重要疾病。HFpEF患者约占总心力衰竭人数的50%以上,且病死率高,预后差。目前关于HFpEF的机制并不清楚,且缺乏有效的治疗药物。对HFpEF病理生理机制的理解既受到人体心肌活检的限制,也受到缺乏完整模拟人类病理的动物模型的限制。因此,建立合适的动物模型有助于深入了解HFpEF的病理生理机制及分子信号通路,并为潜在治疗的临床前研究提供新思路。现综述目前可用于研究HFpEF的动物模型及其优缺点。     

左室射血分数保留的心力衰竭研究进展

心力衰竭是各种临床心血管疾病发展的终末阶段,高发生率及高病死率使其受到越来越多的关注。在美国,其总体发生率约为1.5%～2.0%,而在年龄≥65岁的人群当中,则高达6%～10%[1]。中国的一项随机调查研究选取了15518位成年人(年龄     

肌浆网Ca~(2+)ATP酶在充血性心力衰竭中的研究

充血性心力衰竭(CHF)是各种心脏病终末期的共同归宿,发病率和死亡率均较高。研究表明,肌浆网膜上几个关键调节分子的功能障碍引起心肌细胞内钙调节的内在缺损,在CHF的发病过程中发挥了重要的作用。本文对肌浆网Ca~(2 )ATP酶的蛋白结构、调节机制,在CHF的变化及其在CHF基因治疗方面应用的进展作一综述。     

左心室射血分数保留的心力衰竭患者心率震荡的临床意义研究

目的研究左心室射血分数保留的心力衰竭(heart failure with preserved ejection fraction,HFPEF)患者窦性心率震荡(heart rate turbulence,HRT)的变化以及临床意义。方法选择52例在河南科技大学第一附属医院接受治疗的HFPEF患者作为HEPEF组,另选择无心肌缺血及器质性心脏病变患者30例作为对照组。行24h动态心电图检查,计算HRT指标:震荡初始值(turbulence onset,TO)和震荡斜率(turbulence slope,TS)。行超声心动图检查测定评价心功能不全的相关指标,比较HFPEF组与对照组之间HRT的差异。结果与对照组比较,HFPEF组TO明显升高[(0.17±1.40)%vs(-0.26±0.99)%,P=0.027],TS明显降低[(0.88±2.51)ms/RRI vs(2.60±2.76)ms/RRI,P=0.003]。Pearson相关性分析显示左心室舒张末期容积指数与TO负相关(r=-0.55,P=0.01),与TS呈正相关(r=0.23,P=0.03);舒张早期二尖血流速度和二尖瓣环运动速度比值与TO正相关(r=0.21,P=0.04),与TS呈负相关(r=-0.39,P=0.01)。结论 HFPEF患者窦性心率震荡现象明显受损,提示心脏自主神经调节功能下降,并且反映左心室舒张功能。     

生物标志物在射血分数保留的心力衰竭中的研究进展

心力衰竭是一种复杂的临床综合征,严重危害人类健康。随着对心力衰竭的不断深入研究,人们发现生物标志物在心力衰竭、尤其是在射血分数保留的心力衰竭(HFpEF)中扮演着越来越重要的角色。不同途径(包括神经激素激活、心肌损伤、炎症和纤维化等)的生物标志物对HFpEF具有超出诊断范围的临床效用。本文主要对钠尿肽和新型生物标志物如乳糖凝集素-3及转化生长因子-15等在HFpEF患者诊断及预后中的研究进展进行综述。     

射血分数保留的心力衰竭影像学研究进展

<正>心力衰竭是严重威胁人类健康的高发病率和高致死率的疾病~([1-2])。2016年欧洲心脏学会制定的《急性和慢性心力衰竭诊断和治疗指南》将心力衰竭分为射血分数保留心力衰竭(heart failure with preserved ejection fraction,HFpEF)、射血分数减低心力衰竭(heart failure with reduced ejection fraction,HFrEF)和中间发展期心力衰竭(heart failure with mid-range ejection fraction,HFmrEF)~([3])。其中,HFmr EF是指LVEF介于40%～49%范围的左心功能减低,这一范围被视为心力衰竭发展的"灰色区域"(grey area)。而HFpEF是指由于长时间左心室充盈阻力增加导致的临床综合征,临床定义是LVEF≥50%的心力衰竭。HFpEF患者早期可无明显的临床     

左心房功能与射血分数保留型心力衰竭的研究进展

心力衰竭（HF）是心血管疾病发展的终末阶段,严重影响患者的生存质量及预后。近年来,射血分数保留型心力衰竭（HFpEF）约占HF总人数的50 %,它正日益成为一个全球健康问题。左心房对于调节左室充盈量和维持全心功能至关重要,最近研究发现左心房功能在HFpEF的发生和进展中发挥着重要作用,与HFpEF的不良预后独立相关。因此,该文对HFpEF与左心房结构和功能二者关系展开综述,以期进一步认识左心房结构和功能在HFpEF患者诊断和评估中的重要性。