高血压患者及大鼠线粒体ATPase 6基因的克隆、表达与突变研究

目的：研究正常血压与高血压鼠以及正常血压与高血压患者的差异基因。方法：用差别杂交法（Differentialhybridization）筛选正常人心脏cDNA文库的部分克隆,得到多个在自发性高血压鼠（SHR）心肌和高血压患者外周白细胞中高表达的线粒体基因。重组质粒,PCR－SSCP和测序,进行基因突变分析。结果：SHR鼠心肌线粒体ATPase6基因构象改变,8701位碱基由G→A（G8701A）,编码的第59位氨基酸密码子由丙氨酸变成苏氨酸;A8708G,编码的第61位氨基酸由组氨酸变成精氨酸。高血压病患者外周血白细胞线粒体ATPase6基因8584位碱基突变（G8584A）,编码的第20位氨基酸由丙氨酸变成苏氨酸,这一变化与SHR线粒体ATPase6基因改变相一致。结论：高血压病患者外用血白细胞线粒体ATPase6基因G8584A突变很可能在高血压病心肌肥厚的发生、发展中具有意义,并可能是高血压病的特异性改变。

Cloning, expression and mutation of mitochondrial ATPase gene of hypertensive patients and rats

Objective: To study the different expression of mitochondrial ATPase gene in normotensive and hypertensive human beings and rats. Methods: Partial cDNA clones were screened from human heart cDNA library with differential hybridization. Several mitochondrial genes were highly expressed in the DNA of the cardiac muscle of rats with spontaneous hypertension (SHRs) and in that of peripheral blood leukocytes of patients with hypertension. PCR-SSCP and sequencing were used to study the mutation of the genes.Results: There was polymorphism of mitochondrial ATPase 6 gene in the myocardium of SHR. The base of 8701 changed from G to A (G8701A) and the coden of 59 changed from alanine to threonine and the base of 8708 from A to G and the coden of 61 frorn histidine to arginine. The mitochondrial ATPase 6 gene in the leucocytes of hepertensive patients underwent simi1ar mutations of G8584A and from alanine to threonine in coden 20. Conclusion: The n1utation of G8584A in hypertensive patients is a specific change in hypertension and ! may be responsible for the initiation and development of myocardial hypertrophy.