舒张性心力衰竭患者Ca^(2+)调控蛋白基因转录和蛋白质表达的研究  

Molecular mechanisms underlying calcium handling in diastolic heart failure

作　　者：钟明[1] 张运[2] 张薇[2] 卞继峰[3] 耿昭[3] 赵静[2] 

机构地区：[1]山东医科大学附属医院保健科,济南250012 [2]山东医科大学附属医院心内科,济南250012 [3]山东医科大学分子生物学实验室

出　　处：《中华心血管病杂志》2001年第1期37-40,共4页Chinese Journal of Cardiology

基　　金：国家自然科学基金资助项目 (编号 :3 9970 2 94)

摘　　要：目的　探讨心肌细胞内Ca2 + 超负荷及Ca2 + 调控蛋白在舒张性心力衰竭 (DHF)患者发生中的作用。方法　采用RT PCR和Westernblot技术测定DHF患者和正常对照组Ca2 + 调控蛋白mRNA转录和蛋白质表达的变化。结果　与正常组相比 ,DHF患者心肌细胞内Ca2 + 含量提高 3倍以上 (116 3.88± 5 6 8.6 7vs 2 47.75± 146 .86 ,P <0 .0 1) ;肌浆网 (SR)Ca2 + ATPasemRNA水平减低 33.6 4%(0 .71± 0 .19vs 1.0 7± 0 .11,P <0 .0 5 ) ,细胞膜L型Ca2 + 通道mRNA水平减低 30 .19% (0 .74± 0 .2 0vs1.0 6± 0 .13,P <0 .0 5 ) ,SR磷酸受纳蛋白和兰尼碱受体、肌集钙蛋白的mRNA转录无显著性差异 (P >0 .0 5 ) ;Ca2 + ATPase蛋白的相对含量明显低于对照组 (0 .76± 0 .11vs 1.0 1± 0 .0 3,P <0 .0 5 ) ;磷酸受纳蛋白的相对含量无显著性差异 (0 .98± 0 .0 4vs 1.0 0± 0 .0 2 ,P >0 .0 5 )。结论　SRCa2 + ATPase基因转录和蛋白质表达减低以及细胞膜L型Ca2 + 通道基因转录减低是导致心肌细胞内Ca2 +Objective To elucidate molecular mechanisms underlying calcium handling in patients with diastolic heart failure (DHF) from mRNA level and protein expression. Methods 6 patients with DHF and 6 control subjects were included. The content of calcium was assayed by spectrocomparator. The mRNA amounts of these calcium handling genes including L type calcium channel, sarcoplasmic reticular (SR) Ca 2+ ATPase, ryanodine receptor, calsequestrin and phospholamban were measured by reverse transcription polymerase chain reaction (RT PCR) and normalized to the mRNA levels of actin. The protein levels of SR Ca 2+ ATPase and phospholamban were analyzed by Western blot analysis. Results The content of calcium was significantly increased by 369.78% in patients with DHF. RT PCR analyses showed that the steady state level of mRNA encoding the SR Ca 2+ ATPase and cardiac L type calcium channel were decreased significantly in patients with DHF (0.71±0.19 vs 1.07±0.11, P <0.05, 0.74±0.20 vs 1.06±0.13, P <0.05). In contrast, no alterations of the mRNA levels for SR phospholamban, ryanodine receptor, calsequestrin were observed in patients with DHF compared with those in control subjects. Protein level of SR Ca 2+ ATPase was significantly reduced in patients with DHF (0.76±0.11 vs 1.01±0.03, P <0.05), whereas protein level of phospholamban was unchanged. Conclusion The expressions of cardiac L type calcium channel and SR Ca 2+ ATPase were down regulated in DHF. These changes may be a contributory factor for DHF.

关 键 词：舒张性心力衰竭 钙调蛋白 基因转录 蛋白质 聚合酶链反应 

分 类 号：R541.6[医药卫生—心血管疾病]