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目的通过观察HSP27、HSP70、HSP90在高温、氧化应激的人晶状体上皮细胞中的表达情况,探讨白内障的发病机制。方法体外培养人晶状体上皮细胞,分别在高温(45℃)、氧化(50mmol·L-1H2O2)条件下培养30min后,恢复至正常条件,于不同时间段(0h、2h、4h、6h、16h、24h)采用RTPCR法检测HSP27、HSP70、HSP90的表达情况。结果晶状体上皮细胞在生理和应激情况下均有HSP的表达。热休克和氧化应激后2h导致热休克蛋白mRNA表达明显增加。其中HSP27在2种应激后6h表达最高;HSP70于高温后2h表达最高,以后逐渐降低,而在氧化组中6h表达最高;HSP90在2种应激后4h达到峰值。随后逐渐降低,但它们在16h后仍维持在较高水平。结论晶状体上皮细胞中存在HSP。应激情况下,诱导HSP合成增加,其作为一种对抗应激的蛋白质可能对晶状体上皮细胞起着重要的保护作用。
Objective To investigate the expression of HSP27, HSP70 and HSP90 in human lens epithelial cells under high temperature and oxidative stress and to explore the pathogenesis of cataract. Methods Human lens epithelial cells were cultured in vitro and were returned to normal condition after being incubated at high temperature (45 ℃) and oxidative (50mmol·L-1H2O2) for 30min. The cells were cultured in different time periods (0h, 2h, 4h, 6h, 16h, 24h) RTPCR method was used to detect the expression of HSP27, HSP70 and HSP90. Results The lens epithelial cells both had HSP expression under physiological and stress conditions. Heat shock and oxidative stress 2 h lead to heat shock protein mRNA expression increased significantly. The expression of HSP27 was the highest at 6h after the two stresses. The expression of HSP70 was highest at 2h after high temperature and then gradually decreased at 6h, and reached its peak at 6h in the oxidative group. HSP90 peaked at 4h after the two stresses. Then gradually decreased, but they remained at a high level after 16 hours. Conclusion There is HSP in lens epithelial cells. Stress induces an increase in HSP synthesis, which as an anti-stress protein may play an important protective role in lens epithelial cells.