VARIOUS KINDS OF STRESS AND SMALL HEAT SHOCK PROTEIN, α-CRYSTALLIN mRNA EXPRESSION IN LENS EPITHELIAL CELLS

α-晶体蛋白是一种热休克蛋白，分子量约为27 kd (HSP27)，其耐热性来源于HSP功能。本研究着眼于a-晶状体蛋白在晶状体混浊和晶状体上皮细胞中的作用，研究了手术中获得的晶状体上皮细胞中α-晶状体蛋白mRNA的表达。19只患有年龄相关性白内障的眼睛，以及6只在玻璃体手术中摘除晶状体的年轻患者作为对照。前囊置于Zol试剂中提取上皮细胞RNA。制备cDNA，利用syber green 1实时荧光定量PCR检测a-crystallin mRNA（拷贝数）的表达水平。引物的编码为NM_000394。对照mRNA为GAPDH mRNA，根据α-晶体蛋白mRNA/GAPDH mRNA的归一化比值判断表达水平。年龄相关性白内障归一化比值均值为0.027，对照组为0.187，两组比较差异有统计学意义（t检验，p=0.01）。晶状体上皮细胞α-晶体蛋白表达降低，即HSP功能降低，可能与晶状体混浊的形成有关。

α-crystallin is a heat shock protein with about 27 kd molecular weight (HSP27), and its heat tolerance is derived from HSP function. In this study, focusing on the role of a-crystallin in lens opacity and lens epithelial cells, we investigated mRNA expression of α-crystallin in lens epithelial cells obtained during surgery. Nineteen eyes with age-related cataract, and for the control, Six eyes from which the lens was removed during vitreous surgery in young patients were investigated. Anterior capsule was placed in Zol reagent to extract epithelial cell RNA. Furthermore, cDNA was prepared, and the expression level of a-crystallin mRNA (copies) was measured by real-time PCR using syber green 1. Primers were prepared based on Accession No NM_000394. For the control mRNA, GAPDH mRNA, was used, and the expression level was judged based on the normalized ratio (α-crystallin mRNA/GAPDH mRNA). The mean normalized ratio was 0.027in age-related cataract and 0.187 in the control, with a significant difference between the two groups (t-test, p=0.01). Decreased expression of α-crystallin in lens epithelial cells, i.e., decreased HSP function, may be involved in opacity formation in the lens.