摘要
目的探讨曲安奈德(TA)对体外培养的人视网膜色素上皮(RPE)细胞热休克蛋白47(HSP47)表达的影响。方法研究RPE细胞分别在0、50、200μg/mLTA的培养条件下,通过RT-PCR检测HSP47 mRNA的表达,使用Western blot检测其蛋白水平。结果与0μg/mLTA组比较,50μg/mL及200μg/mLTA组HSP47 mRNA和蛋白的表达量明显下降(P<0.05),蛋白水平亦明显下降(P<0.05)。结论HSP47对胶原的成熟起着重要作用,TA能有效抑制人RPE细胞HSP47的表达,可能是其防治增生性玻璃体视网膜病变(PVR)及增生型糖尿病视网膜病变(PDR)的一个重要机制。
Objective Triamcinolone acetonide （TA） may offer a possibility for adjunctive treatment of intraocular proliferative diseases. However,the underlying molecular mechanisms responsible for treatment of intraocular proliferative diseases is not fully understood. This experiment was performed to explore the effect of TA on the production of heat shock protein 47 （HSP47） in cultured human retinal pigment epithelial （RPE）cells. Methods Human eyeballs were obtained from donors within 8 hours after death. Eyeballs were digested by trypin and RPE cells were primarily cultured in DMEM containing 10% newborn calf serum,25 mmol/L glucose, 100 U/mL penicillin and 100 μg/mL streptomycin. The 3th to 5th generation of cells were consequently cultured in DMEM containingl0% new-born calf serum,25 mmol/L glucose, and 0,50 and 200 μg/mL TA was added in culture medium respectively. RT-PCR was used to examine the expression of HSP47 mRNA,and Western blot analysis was used to measure the levels of HSP47 protein. Results The mRNA expression and protein synthesis of HSP47 in human RPE cells was gradually decreased with the increase of TA concentration （ F = 16. 456, P = 0. 000; F = 10. 004, P = 0. 001 respectively）. The AHSP47/Aβ-actin and AhsP47/protein were obviously down-regulated in 50 and 200 μg/mL TA group compared with 0μ g/mL TA group （P 〈 0.05 ）. Conclusion HSP47 plays a specific role in the intracellular processing of procollagen production as a collagen-specific molecular chaperone. TA has a potential effect in the treatment of intraocular proliferative diseases by down-regulating HSP47 in human RPE cells.
出处
《眼科研究》
CSCD
北大核心
2008年第5期369-372,共4页
Chinese Ophthalmic Research
关键词
曲安奈德
热休克蛋白47
人视网膜色素上皮细胞
triamcinolone acetonide
heat shock protein 47
human retinal pigment epithelial cell
