摘要
目的探讨曲安奈德(triamcinolone acetonide,TA)对体外缺氧培养的人视网膜色素上皮(retinal pigment epithelium,RPE)细胞热休克蛋白47(heat shock protein47,HSP47)表达的影响。方法使用化学缺氧诱导剂CoCl2模拟人RPE细胞缺氧环境,人RPE细胞分别在不同浓度TA的条件下培养,将其分为0μg.L-1TA组、0.05μg.L-1TA组和0.20μg.L-1TA组,均加入含150μmol.L-1CoCl2的DMEM完全培养液采用RT-PCR检测HSP47表达,使用Western-blotting检测HSP47的蛋白水平。结果 0μg.L-1TA组、0.05μg.L-1TA组和0.20μg.L-1TA组HSP47mRNA的表达量分别为1.53±0.21、1.15±0.20、1.07±0.17,HSP蛋白表达量分别为748.62±79.03、643.76±63.18、615.62±61.77,3组间HSP47mRNA及蛋白表达量差异均有统计学意义(F=14.729,P=0.000;F=9.438,P=0.001);0.05μg.L-1TA组与0μg.L-1TA组比较,HSP47mRNA及蛋白的表达量均显著下降(P=0.000、P=0.003);0.20μg.L-1TA组与0μg.L-1TA组比较,HSP47mRNA及蛋白的表达量也均显著下降(均为P=0.000);而0.05μg.L-1TA组与0.20μg.L-1TA组比较,HSP47mRNA及蛋白的表达量差异均无统计学意义(P=0.373、P=0.392)。结论 HSP47对胶原的成熟起着重要的作用,TA能有效地抑制缺氧条件下人RPE细胞HSP47的表达,可能是其防治增生性玻璃体视网膜病变的一个重要机制。
Objective To investigate effect of triamcinolone acetonide （TA） on heat shock protein 47 （ HSP47 ） in cultured human retinal pigment epithelium （ RPE ） cells under anoxia condition. Methods Chemical hypoxic revulsive CoCl2 was used to imitate the anoxia condition of RPE cells. Human RPE cells were cultured in different concentrations of TA and divided into 0 ug · L-1, 0.05 ug · L-1 and 0.20 ug · L-1 TA groups. RT-PCR was used to detect the expression of HSP47 mRNA, and Western-blotting was used to measure levels of HSP47 protein. Results In 0 ug · L-1, 0.05 ug · L-1 and 0.20 ug · L-1 TA groups,expression levels of HSP47 mRNA were 1.53 ±0.21, 1. 15 ± 0.20 and 1.07 ± 0. 17, HSP47 protein levels were 748.62 ± 79.03,643.76 ± 63. 18 and 615.62 ± 61.77, respectively. There were significant differences in HSP47 mRNA and protein levels among groups （F = 14. 729 ,P = 0. 000;F = 9.438 ,P = 0. 001 ） ; Compared with 0 ug ·L- 1 TA group, HSP47 mRNA and protein level were obviously lower than those in 0.005 ug · L-1 TA group（P =0.000,P =0.003） ,also lower in 0.20 ug · L-1 TA group （ both P = 0. 000）. While no statistical difference was found in HSP47 mRNA and protein level between 0.05 ug · L-1 TA group and 0.20 ug · L-1 TA group（P =0.373 ,P = 0.392）. Conclusions HSP47 plays an important role in maturation of collagen. TA can effectively inhibit expression of HSP 47 in human RPE cells in anoxia condition, which may be an important mechanism of preventing proliferative vitreoretinopathy.
出处
《眼科新进展》
CAS
北大核心
2010年第8期737-739,744,共4页
Recent Advances in Ophthalmology
关键词
曲安奈德
缺氧
热休克蛋白47
视网膜色素上皮细胞
triamcinolone acetonide
hypoxia
heat shock protein 47
retinal pigment epithelial cell
