摘要
为了研究Ⅱ型糖尿病（ＮＩＤＤＭ）患者红细胞膜血型糖蛋白Ａ（ＧＰＡ）的基因表达，采用不连续聚蔗糖（Ｐｅｒｃｏｌ）密度梯度法分离１５名正常人和２５例ＮＩＤＤＭ患者外周血的网织红细胞，提取其总ＲＮＡ，继以Ｎｏｒｔｈｅｒｎ斑点杂交观测此两组的红细胞膜ＧＰＡ基因的表达水平．采用碱性磷酸酶酶联免疫检测完成红细胞膜ＧＰ的Ｗｅｓｔｅｒｎ印迹．结果表明，ＮＩＤＤＭ患者组红细胞膜ＧＰＡ的ｍＲＮＡ含量较正常人组显著增加（分别为１１．９２±１０．５和１０．１８±１．０８积分光密度，Ｐ＜０．０１）．免疫印迹图谱显示，有６例ＮＩＤＤＭ患者的区带３ａ、３ｂ界限模糊，３例的区带３ａ、３ｂ和４明显浅染．推测ＮＩＤＤＭ患者红细胞膜ＧＰＡ减少可能引起其基因表达呈代偿性增强，而障碍或许出现在翻译环节上，这些与ＮＩＤＤＭ患者红细胞变形能力（ＲＣＤ）的明显降低甚为相符．
To study the gene expression of erythrocyte membrane glycophorin A(GPA)in patients with non insulin dependent diabetes mellitus(NIDDM),human reticulocytes were separated by discontinuous density gradient of percoll from peripheral blood of 15 healthy subjects and 25 NIDDM patients,and total RNAs of their reticulocytes were isolated.Using the reticulocyte RNAs,the gene expression levels of membrane GPA were estimated by Northern dot hybridization.Western blotting was performed by alkaline phosphatase linked immunodetection.The contents of GPA mRNA in NIDDM group were significantly increased than those of the control group(11 92±1 05 vs.10 18±1 08 integrated OD ,respectively; P <0 01).The Western immunoblots of erythrocyte membrane GP revealed ill defined demarkation between bands 3a and 3b in 6 NIDDM patients and weakly stained bands of 3a,3b and 4 in other 3 patients.Therefore,it is inferred that the reduction of erythrocyte membrane GPA in NIDDM may promote the compensatory elevation of its gene expression,and the aberration would occur in related translation step.This deduction is possibly in accordance with the findings of distinct decrease of red cell deformability (RCD) in NIDDM patients.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1998年第5期622-626,共5页
Chinese Journal of Biochemistry and Molecular Biology
关键词
糖尿病
GPA
基因表达
Diabetes mellitus,Non insulin dependent(NIDDM),Erythrocyte membrane,Glycophorin A,Gene expression
