肌浆网钙ATP酶过表达对心房颤动兔心房及L型钙通道a1c蛋白表达的影响
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国家国际科技合作专项(2011DFA32860)


Effects of sarcoendoplasmic reticulum Ca2 -ATPase2a overexpression on the protein expression of L-type voltage dependent calcium channel α1c subunit in a rabbit model of acute atrial fibrillation
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    摘要:

    目的 探讨心肌肌浆网钙ATP酶2a(SERCA2a)基因过表达,对兔急性心房颤动(AF)的心房肌L型电压依赖钙通道蛋白alc亚单位(LVDCCalc)表达的影响。方法 48只成年健康新西兰大白兔,随机分成为房颤组(AF组,n=12)、9-型腺相关病毒 增强型绿色荧光蛋白组 房颤组(rAAV9 EGFP AF组,n=12)、9-型腺相关病毒 肌浆网钙ATP酶2a 增强型绿色荧光蛋白组 房颤组(rAAV9 SERCA2a EGFP AF组,n=12),并设假手术组作为对照(Control组,n=12)。rAAV9 EGFP AF组和rAAV9 SERCA2a EGFP AF组起搏前30d开胸心包腔内注射包含报告基因的rAAV9-EGFP和包含靶基因的rAAV9-SERCA2a-EGFP各500祃L(1?012v.g/礚)。转染30 d后,AF组、rAAV9 EGFP AF组和rAAV9 SERCA2a EGFP AF组,以600BPM刺激频率,持续起搏兔右心房24小时制作AF模型。处死各组动物,倒置荧光显微镜下观察右心房肌组织EGFP表达情况,行HE染色对右心房组织形态检查,应用免疫组化技述检测SERCA2a蛋白及LVDCCalc蛋白在心房肌中的表达情况。结果rAAV9 EGFP SERCA2a AF组SERCA2a蛋白及LVDCCalc蛋白表达量显著高于AF组和rAAV9 EGFP AF组(P<0.05),但与对照组比较无显著差异。结论 心房肌转SERCA2a基因,显著减少了急性房颤心房肌的LVDCCa1c蛋白表达的降低,有逆转急性AF电重构的作用。国家国际科技合作专项的编号2011DFA32860

    Abstract:

    【Abstract】 0bjective To explore the effects of sarcoendoplasmic reticulum calcium ATPase2a(SERCA2a) overexpression on the protein expression of L-type voltage dependent calcium channel (LVDCCalc) subunit in the rabbit model of atrial fibrillation(AF). Methods Forty-eight New Zealand rabbits were randomized into four groups (n=12),including control group,AFgroup,rAAV9 EGFP AFgroupand rAAV9 SERCA2a EGFP AF group.rAAV9-EGFP and rAAV9-SERCA2a-EGFP(1.0?012v.g/礚 for each animal )were injected into pericardium of the animals of rAAV9 EGFP AF and rAAV9 SERCA2a (enhanced green fluorescent protein)EGFP AF groups before rapid atrial pacing thirty day. Thirty day after gene transfection,AF was induced by rapid atrial pacing(600 beats/min)for 24 hours in each animal of AF group,rAAV9 EGFP AF group and rAAV9 SERCA2a EGFP AF group .Each animal of control group was not paced,Animals were scarified to observe expression of the green fluorescent protein in the myocardial frozen section of rabbits in rAAV9 EGFP AF group under the inverted fluorescence microscope,to assess the right atrial tissue morphology and detect the expressions of SERCA2a protein and LVDCCalc protein in the right atrial tissue in each group by H E staining and immunohistochemistry at 24 hours after pacing. Results Compared with AF group and rAAV9 EGFP AF group(P<0.05),expressions of SERCA2a protein and LVDCCalc protein in the right atrium of rabbits in rAAV9 SERCA2a EGFP AF group was significantly upregulated,but there is no difference on expressions of SERCA2a protein and LVDCCalc protein in the right atrium of rabbit among rAAV9 SERCA2a EGFP AF group,AF group and rAAV9 EGFP AF group (P>0.05). Conclusions The atrial tissue transferred SERCA2a protein could significantly decrease the downregulation of the LVDCCalc protein expression in this rabbit model of AF and reverse acute AF electrical remodeling.

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张广伟,张 玲,王凌鹏,徐晓霞,张峰,杨曦,马依彤,侯月梅.肌浆网钙ATP酶过表达对心房颤动兔心房及L型钙通道a1c蛋白表达的影响[J].中国实验动物学报,2013,21(4).

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  • 收稿日期:2013-01-11
  • 最后修改日期:2013-02-17
  • 录用日期:2013-02-21
  • 在线发布日期: 2013-08-30
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