Chertow B S, Blaner W S, Rajan N, Primerano D A, Meda P, Cirulli V, Krozowski Z, Smith R, Cordle M B
Medical Service, Veterans Administration Medical Center, Huntington, West Virginia.
Diabetes. 1993 Aug;42(8):1109-14. doi: 10.2337/diab.42.8.1109.
To define the mechanism of vitamin A action at the beta-cell level, we tested for the presence of messenger RNA for retinoic acid receptors alpha, beta, and gamma; cytosolic retinol-binding protein; and cytosolic retinoic acid-binding protein in RINm5F cells, an insulin-secreting cell line, and determined whether cytosolic retinol-binding protein and cytosolic retinoic acid-binding protein are present in isolated purified normal rat beta-cells. Northern blot analyses showed two transcripts of retinoic acid receptor alpha messenger RNA (3.8 and 2.4 kb), one transcript of retinoic acid receptor messenger RNA (3.8 kb), and one transcript of cytosolic retinol-binding protein (0.9 kb) in RINm5F cells. Ribonuclease protection assays also showed the presence of cytosolic retinol-binding protein and cytosolic retinoic acid-binding protein in RINm5F cells. Quantitatively, cytosolic retinol-binding protein levels were 0.10 +/- 0.02 pg/micrograms total RNA. Using specific radioimmunoassays, normal isolated purified rat beta-cells contained CRBP (19.2 +/- 2.38) and cytosolic retinoic acid-binding protein (16 +/- 0.53 ng/10(6) cells). The presence of message for retinoic acid receptors alpha and gamma, cytosolic retinol-binding protein, cytosolic retinoic acid-binding protein, and the gene products of cytosolic retinol-binding protein and cytosolic retinoic acid-binding protein in insulin-secreting cells support a mechanism of vitamin A action and role for cytosolic and nuclear receptors at the beta-cell level similar to that suggested in nonendocrine cells.(ABSTRACT TRUNCATED AT 250 WORDS)
为了确定维生素A在β细胞水平的作用机制,我们检测了胰岛素分泌细胞系RINm5F细胞中视黄酸受体α、β和γ的信使核糖核酸、胞质视黄醇结合蛋白和胞质视黄酸结合蛋白的存在情况,并确定分离纯化的正常大鼠β细胞中是否存在胞质视黄醇结合蛋白和胞质视黄酸结合蛋白。Northern印迹分析显示,RINm5F细胞中有视黄酸受体α信使核糖核酸的两种转录本(3.8和2.4 kb)、视黄酸受体信使核糖核酸的一种转录本(3.8 kb)以及胞质视黄醇结合蛋白的一种转录本(0.9 kb)。核糖核酸酶保护分析也显示RINm5F细胞中存在胞质视黄醇结合蛋白和胞质视黄酸结合蛋白。定量分析显示,胞质视黄醇结合蛋白水平为0.10±0.02 pg/μg总核糖核酸。使用特异性放射免疫测定法,正常分离纯化的大鼠β细胞含有CRBP(19.2±2.38)和胞质视黄酸结合蛋白(16±0.53 ng/10⁶细胞)。胰岛素分泌细胞中视黄酸受体α和γ的信使、胞质视黄醇结合蛋白、胞质视黄酸结合蛋白以及胞质视黄醇结合蛋白和胞质视黄酸结合蛋白的基因产物的存在,支持了维生素A的作用机制以及β细胞水平上胞质和核受体的作用,这与非内分泌细胞中所提示的类似。(摘要截短于250词)