国际眼科杂志
國際眼科雜誌
국제안과잡지
INTERNATIONAL JOURNAL OF OPHTHALMOLOGY
2014年
5期
811-814
,共4页
王业青%王健%吕勇%董丽%穆华%曹文萍
王業青%王健%呂勇%董麗%穆華%曹文萍
왕업청%왕건%려용%동려%목화%조문평
整联蛋白连接激酶%视网膜色素上皮细胞%缺氧%热休克蛋白90%17-丙烯胺基-17-去甲氧基格尔德霉素
整聯蛋白連接激酶%視網膜色素上皮細胞%缺氧%熱休剋蛋白90%17-丙烯胺基-17-去甲氧基格爾德黴素
정련단백련접격매%시망막색소상피세포%결양%열휴극단백90%17-병희알기-17-거갑양기격이덕매소
integrin- linked kinase%retinal pigment epithelium cells%hypoxia%heat shock protein 90%17-allylamio-17-demethoxygeldanamycin
目的：研究热休克蛋白(heat shock protein 90,Hsp90)抑制剂17-AAG对缺氧诱导的视网膜色素上皮( retinal pigment epithelial,RPE)细胞整联蛋白连接激酶( integrin - linked kinase,ILK)表达的影响。<br> 方法：利用200μmol/L氯化钴( cobalt chloride, CoCl2)建立体外RPE细胞的化学缺氧模型。不同浓度的Hsp90抑制剂17-AAG预处理RPE细胞1 h后给予12 h缺氧处理。实验分为缺氧对照组、二甲基亚砜( DMSO)对照组和17-AAG预处理组。其中17-AAG预处理组根据浓度不同又分为6组：0.01,0.10,0.50,1.00,5.00,10.00μmol/L。应用RT-PCR和Western blot 方法检测ILK表达变化情况。<br> 结果：缺氧对照组、DMSO对照组和17-AAG预处理组ILK mRNA与内参β-actin mRNA 光密度比值分别为1.32依0.04,1.29依0.03,0.93依0.06,0.70依0.05,0.53依0.03,0.44依0.04,0.32依0.04,0.30依0.03；ILK 蛋白与内参β-actin蛋白光密度比值分别为2.16依0.04,2.13依0.04,1.65依0.04,1.13依0.05,0.74依0.03,0.41依0.06,0.35依0.04,0.35依0.03。与缺氧对照组比较,17-AAG预处理组ILK mRNA和蛋白的表达明显降低( P<0.05),呈浓度依赖性。<br> 结论：Hsp90抑制剂17-AAG能够降低缺氧条件下RPE细胞ILK的表达。
目的：研究熱休剋蛋白(heat shock protein 90,Hsp90)抑製劑17-AAG對缺氧誘導的視網膜色素上皮( retinal pigment epithelial,RPE)細胞整聯蛋白連接激酶( integrin - linked kinase,ILK)錶達的影響。<br> 方法：利用200μmol/L氯化鈷( cobalt chloride, CoCl2)建立體外RPE細胞的化學缺氧模型。不同濃度的Hsp90抑製劑17-AAG預處理RPE細胞1 h後給予12 h缺氧處理。實驗分為缺氧對照組、二甲基亞砜( DMSO)對照組和17-AAG預處理組。其中17-AAG預處理組根據濃度不同又分為6組：0.01,0.10,0.50,1.00,5.00,10.00μmol/L。應用RT-PCR和Western blot 方法檢測ILK錶達變化情況。<br> 結果：缺氧對照組、DMSO對照組和17-AAG預處理組ILK mRNA與內參β-actin mRNA 光密度比值分彆為1.32依0.04,1.29依0.03,0.93依0.06,0.70依0.05,0.53依0.03,0.44依0.04,0.32依0.04,0.30依0.03；ILK 蛋白與內參β-actin蛋白光密度比值分彆為2.16依0.04,2.13依0.04,1.65依0.04,1.13依0.05,0.74依0.03,0.41依0.06,0.35依0.04,0.35依0.03。與缺氧對照組比較,17-AAG預處理組ILK mRNA和蛋白的錶達明顯降低( P<0.05),呈濃度依賴性。<br> 結論：Hsp90抑製劑17-AAG能夠降低缺氧條件下RPE細胞ILK的錶達。
목적：연구열휴극단백(heat shock protein 90,Hsp90)억제제17-AAG대결양유도적시망막색소상피( retinal pigment epithelial,RPE)세포정련단백련접격매( integrin - linked kinase,ILK)표체적영향。<br> 방법：이용200μmol/L록화고( cobalt chloride, CoCl2)건입체외RPE세포적화학결양모형。불동농도적Hsp90억제제17-AAG예처리RPE세포1 h후급여12 h결양처리。실험분위결양대조조、이갑기아풍( DMSO)대조조화17-AAG예처리조。기중17-AAG예처리조근거농도불동우분위6조：0.01,0.10,0.50,1.00,5.00,10.00μmol/L。응용RT-PCR화Western blot 방법검측ILK표체변화정황。<br> 결과：결양대조조、DMSO대조조화17-AAG예처리조ILK mRNA여내삼β-actin mRNA 광밀도비치분별위1.32의0.04,1.29의0.03,0.93의0.06,0.70의0.05,0.53의0.03,0.44의0.04,0.32의0.04,0.30의0.03；ILK 단백여내삼β-actin단백광밀도비치분별위2.16의0.04,2.13의0.04,1.65의0.04,1.13의0.05,0.74의0.03,0.41의0.06,0.35의0.04,0.35의0.03。여결양대조조비교,17-AAG예처리조ILK mRNA화단백적표체명현강저( P<0.05),정농도의뢰성。<br> 결론：Hsp90억제제17-AAG능구강저결양조건하RPE세포ILK적표체。
AIM: To investigate the effect of 17-allylamio-17-demethoxygeldanamycin ( 17 - AAG ) , an heat shock protein 90(Hsp90) inhibitor, on the expression of integrin-linked kinase ( ILK) induced by hypoxia in retinal pigment epithelium ( RPE) cells. <br> METHODS: RPE cells were cultured with 200μmol/L cobalt chloride ( CoCl2 ) for 12 hours to imitate chemical hypoxia. Pretreatment of 17-AAG was 1 hour prior to hypoxic insult. Experiment was divided into hypoxic control group, Dimethyl sulfoxide ( DMSO) control group and 17-AAG pretreatment group (0. 01, 0. 10, 0. 50, 1. 00, 5. 00 and 10. 00μmol/L ). RT - PCR and Western blot analysis were used to examine the expression of ILK in cultured RPE cells. <br> RESULTS:The density ratios of ILK mRNA and β-actin mRNA of hypoxic control group, DMSO control group and 17-AAG pretreatment groups were 1. 32±0. 04,1. 29±0. 03, 0. 93±0. 06,0. 70±0. 05,0. 53±0. 03,0. 44±0. 04,0. 32±0. 04,0. 30±0. 03;and the density ratios of ILK protein and β-actin protein were 2. 16±0. 04, 2. 13±0. 04, 1. 65±0. 04, 1. 13±0. 05, 0. 74±0. 03, 0. 41± 0. 06, 0. 35± 0. 04, 0. 35± 0. 03. The ILK expression in 17-AAG pretreated groups were inhibited compared to hypoxic control group (P < 0. 05), and the decrease was in concentration-dependent manner. <br> CONCLUSION:Hsp90 inhibitor 17-AAG was effective to inhibit the expression of ILK induced by hypoxia in RPE cells.
