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

各种应激及小热休克蛋白、α-晶状体蛋白在晶状体上皮细胞中的表达

• 批准号: 15591878
• 负责人: USUI Masahiko
• 金额: $ 1.79万
• 依托单位: Tokyo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591878/
• 关键词: α-crystallin; lens epithelial cells; cataract; heat shock protein; 熱ショック蛋白質(HSP27); 水晶体上皮細胞; 水晶体混濁; mRNAの発現

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.

α-晶状体蛋白是一种分子量约为27 kd的热休克蛋白（HSP 27），其耐热性来源于HSP功能。在本研究中，我们研究了α-晶状体蛋白在透镜混浊和透镜上皮细胞中的作用，并在手术过程中获得的透镜上皮细胞中研究了α-晶状体蛋白的mRNA表达。对19例年龄相关性白内障患者和6例在玻璃体手术中摘除透镜的年轻患者进行了研究。将前囊置于Zol试剂中提取上皮细胞RNA。此外，制备cDNA，并使用syber绿色1通过实时PCR测量α-晶状体蛋白mRNA的表达水平（拷贝）。基于登录号NM_000394制备引物。对于对照mRNA，使用GAPDH mRNA，并且基于标准化比率（α-晶状体蛋白mRNA/GAPDH mRNA）判断表达水平。年龄相关性白内障的平均标准化比率为0.027，对照组为0.187，两组之间存在显著差异（t检验，p=0.01）。透镜上皮细胞中α-晶状体蛋白表达减少，即，HSP功能降低可能与透镜中混浊的形成有关。