葡萄糖调节蛋白GRP78在小梁网细胞中表达及临床意义.docVIP

葡萄糖调节蛋白GRP78在小梁网细胞中表达及临床意义 　　[摘要] 目的 检测葡萄糖调节蛋白78（GRP78）在体外培养的正常人小梁网细胞（NTM）及原发性开角型青光眼（POAG）患者小梁网细胞（GTM）中蛋白及mRNA表达差异及临床意义。 方法 将体外培养的NTM及GTM细胞分为衣霉素（Tm）组、十字孢碱（STS）组及对照组，采用Real-time PCR、Western blot方法检测药物处理6、24 h后小梁网细胞中GRP78 mRNA及蛋白表达。 结果 原代培养的GTM细胞中GRP78表达水平明显低于NTM细胞，差异有统计学意义（P 0.05）。Tm可增加NTM、GTM细胞中GRP78的表达，但后者的增高水平明显低于前者，差异有统计学意义（P 0.05）。GTM细胞对STS的反应大于NTM细胞。 结论 POAG患者小梁网细胞对内质网应激反应能力低下，GRP78表达明显下调，对损伤性刺激的敏感性增强，提示GRP78蛋白可能在内质网应激诱导产生的小梁细胞凋亡中起保护作用。 　　[关键词] 青光眼；小梁网细胞；内质网应激 　　[中图分类号] R775 [文献标识码] A [文章编号] 1673-7210（2016）01（c）-0114-04 　　Expression and clinical significance of GRP78 in human trabecular meshwork cells 　　CHAI Fang1 ZHUO Yehong2 YANG Xinguang1▲ 　　1.Department of Ophthalmology， Xian No.4 Hospital Ophthalmic Medical Center of Shaanxi Province Guangren Hospital Affiliated to School of Medicine， Xian Jiaotong University， Shaanxi Province， Xian 710004， China； 2.Zhongshan Ophthalmic Center， Sun Yat-Sen University State Key Laboratory of Ophthalmology， Guangdong Province， Guangzhou 510060， China 　　[Abstract] Objective To detect protein and mRNA expression differences of GRP78 in human trabecular meshwork cells of normal eyes （NTM） and POAG patients （GTM） in vitro. Methods Human trabecular meshwork cells from NTM and GTM in vitro were divided into 3 groups： Tunicamycin （Tm） group， Staurosporine （STS） group and control group. Real-time RT-PCR and Western blot were used to detect the expression of mRNA and protein of GRP78 in trabecular meshwork cells after 6， 24 h treatment. Results The expression of GRP78 in primary culture GTM cells was significantly lower than that in NTM cells， with statistically significant difference （P 0.05）. Tm could increase the expression of GRP78 in NTM and GTM cells， the expression in GTM cells was lower than that in NTM cells， with statistically significant difference （P 0.05）. The response of GTM cells for STS was bigger than NTM cells. Conclusion GTMs showed insensitivity reactions when stimulated with inducer of endoplasmic ret