缺氧对体外培养的人视网膜色素上皮细胞热休克蛋白47表达的影响

Effects of hypoxia on the expression of human retinal pigment epithelium cells cultured in vitro

目的:探讨缺氧对体外培养的人视网膜色素上皮(retinalpigmentepithelium,RPE)细胞热休克蛋白47(heatshockprotein47,HSP47)表达的影响。方法:培养的人RPE细胞分为两组:缺氧组使用化学缺氧诱导剂CoCl2模拟RPE细胞缺氧环境来培养人RPE细胞,对照组置入正常环境中培养。采用半定量RT-PCR检测人RPE细胞中HSP47mRNA的表达,Westernblot蛋白印迹分析检测人RPE细胞中蛋白水平。结果:缺氧组AHSP47/Aβ-actin比值为1.46±0.15与对照组比值0.94±0.13比较,差异有显著意义(P<0.05)。同时缺氧组HSP47蛋白含量平均A值为728.03±38.15显著高于正常对照组平均A值571.47±26.95,差异有统计学意义(P<0.05)。结论:HSP47在缺氧状态下异常高表达。

AIM： To explore the effects of hypoxia on the expression of heat shock protein 47 （HSP47） in cultured human retinal pigment epithelium （RPE） cells. METHODS： Cultured human RPE cells were divided into two groups. One was cultured under hypoxia chamber, the other was cultured under normal chamber. The hypoxia environment was set up with chemical hypoxia inducer-CoCl2. The expression of HSP47 mRNA was determined by semi-quantitative reverse transcription-polymerase chain reaction （RT-PCR）. The expression of HSP47 protein in two groups was detected by western blot analysis. RESULTS： The value of AHSP47/Aβ-actin in the hypoxia group was 1.46± 0.15. The difference between the hypoxia group and that of control group （0.94± 0.13） was statistically significant （P 〈0.05）. Compared with control group （Mean A 571.47± 26.95）, the expression of HSP47 protein in hypoxia group （Mean A 728.03± 38.15） was significant high （P 〈 0.05）.CONCLUSION： The HSP47 is highly expressed in hypoxia cendition, and it may play an important role in the cellagen deposition of proliferative vitroretinopathy.