腺病毒载体介导的人β_2肾上腺素能受体基因在心肌细胞特异性表达的研究  

The study of specific expression of β_2-AR gene in cardiomyocytes infected by adenovirus vector

作　　者：高文谦[1] 李小鹰[1] 李梁[2] 裴雪涛[2] 

机构地区：[1]北京解放军总医院老年心内科,100853 [2]军事医学科学院,国家生物医学分析中心

出　　处：《中华心血管病杂志》2001年第8期494-496,共3页Chinese Journal of Cardiology

基　　金：国家 973基金资助项目 (G2 0 0 0 0 5 6 90 6 )

摘　　要：目的　观察构建的重组腺病毒载体介导的人 β2 肾上腺素能受体 (β2 AR)基因能否在心肌细胞中特异性表达。方法　构建含心肌肌凝蛋白轻链 (mlc 2v)启动子携带 β2 AR基因的重组腺病毒载体 (Admlcβ2 AR) ,用重组病毒感染大鼠心肌细胞、3T3细胞、HFCL细胞及HeLa细胞 ,检测感染的细胞中人β2 AR基因的表达。结果　RT PCR显示只有感染的心肌细胞表达人 β2 ARmRNA ;Western免疫印迹杂交分析表明感染的心肌细胞有人 β2 AR表达 ;放射性配基检测显示感染的心肌细胞 β AR密度 (15 3± 5 .2 )fmol/mg蛋白 ,明显高于未感染的心肌细胞 (76± 2 .8)fmol/mg蛋白 ,P <0 .0 1)。结论构建的重组腺病毒载体可有效地将 β2Objective To establish an adenovirus vector and make β 2 adrenergic receptor (β 2 AR) gene express specificly in cardiomycytes. Methods Recombinant adenovirus (Admlc β 2 AR)carrying the β 2 AR gene under the control of the ventricle specific myosin light chain 2(mlc 2v)promotor was constructed, which infected neonate rat ventricular myocytes,3T3 cells,HFCL cells and HeLa cells in vitro. The expression of β 2 AR gene was assessed. Result The presence of β 2 AR mRNA was demonstrated by RT PCR in infected neonate rat ventricular myocytes in vitro but not in infected 3T3 cells, HFCL cells and HeLa cells . The expression of the β 2 AR gene was demonstrated by Western blot in infected neonate rat ventricular myocytes .According to a ligand binding assay,the density of β AR in the infected cardiomyocytes was greater than that in control [(153±5.2)fmol/mg protein vs (76±2.8) fmol/mg protein, P <0.01]. Conclusion The established adenovirus vector Admlc β 2 AR could allow heart specific expression of β 2 AR gene.

关 键 词：腺病毒载体 心肌细胞 特异性表达 心力衰竭 肾上腺素能受体基因 

分 类 号：R541.6[医药卫生—心血管疾病]