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姜黄素干预人视网膜色素上皮细胞钙离子信号的改变与整合素基因表达的探讨分析
  • ISSN号:1007-9572
  • 期刊名称:《中国全科医学》
  • 时间:0
  • 分类:R774.1[医药卫生—眼科;医药卫生—临床医学]
  • 作者机构:[1]河北医科大学第二医院眼科,河北省石家庄市050000
  • 相关基金:国家自然科学基金资助项目(30973252)
中文摘要:

目的探讨视网膜色素上皮细胞(RPE)的钙离子与整合素基因表达的相关性。方法人RPE细胞经分离培养,免疫细胞化学鉴定后,传至第4~6代;对各组RPE细胞进行5p,mol/LFluo-3负载30rain,姜黄素分别以0.9375、1.8750、3.7500、7.5000、15.0000mg/L浓度干预人RPE细胞,利用激光扫描共聚焦显微镜(LSCM)测定各组RPE细胞内早期钙离子的变化。利用MTT法测定不同浓度的姜黄素干预人RPE细胞48h后的IC50,并观察相应的形态学变化。用Real—TimePCR测定姜黄素干预后的整合素基因的表达。结果LSCM检测结果:空白对照组、0.9375、1.8750、3.7500、7.5000、15.00130mg/L姜黄素组RPE细胞内均有钙荧光。空白对照组钙荧光强度明显低于姜黄素各组,且差异有统计学意义(P〈0.05),对照组基因表达量与整合素甜和整合素β5呈正效应,与整合素α3和整合素α5呈负效应。结论体外培养的人RPE细胞经不同浓度姜黄素干预后,较空白对照组早期细胞内钙离子荧光强度明显增加,且有显著性差异。当细胞受到姜黄素外界信号刺激时,细胞内钙离子浓度能迅速增加,继而调节整合素的活性,继而引起细胞的生物学效应。因此,姜黄素通过调节细胞内的信号传导,可能成为治疗人眼疾,如增殖性玻璃体视网膜病变的一种有效物质。

英文摘要:

Objective To explore the association between calcium signal of retinal pigment epithelial cell (RPE) and the integrin responses. Methods The RPE cells were separated and cultured to the 4 - 6 generations after determination by immunocytochemistry. RPE cells were treated by Fluo -3 (5μmol/L) for 30 min. Each group of RPE ceils was affected with curcumin of 0. 9375 mg/L, 1. 8750 mg/L, 3. 7500 mg/L, 7. 5000 mg/L, and 15. 000 mg/L, respectively. Intracellular calcium changes of RPE cells were measured by laser scanning confocal microscope (LSCM). The RPE cells'IC50 after 48 h incubation in each group was recorded by MTY and related morphology changes were observed. The integrin gene expression was measured by Real-time PCR. Results LSCM results showed calcium fluorescence in RPE cells in the control group and all curcumin groups. The calcium fluorescence in the curcumin groups was significantly greater than that in the control group ( P 〈 0.05 ). The gene expression level was positively correlated with integrin α4 and β5, and negatively correlated with integrin α3 and α5. Conclusion The intracellular calcium fluorescence of curcumin affected human RPE ceils is stronger than that of those not affected. The intracellular calcium immediately increases when stimulated by external curcumin, which can regulate integrin gene expression and lead to biological effects. Therefore, curcumin might be a therapeutic agent to modulate the cell signal transduction for certain human ocular disorders such as PVR.

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期刊信息
  • 《中国全科医学》
  • 北大核心期刊(2011版)
  • 主管单位:中华人民共和国国家卫生和计划生育委员会
  • 主办单位:中国医院协会 中国全科医学杂志社
  • 主编:韩建军
  • 地址:北京市西城区广义街5号广益大厦A座907
  • 邮编:100053
  • 邮箱:zgqkyx@chinagp.net.cn
  • 电话:010-63052088 83525550
  • 国际标准刊号:ISSN:1007-9572
  • 国内统一刊号:ISSN:13-1222/R
  • 邮发代号:80-258
  • 获奖情况:
  • 获2000-2001年度河北省优秀期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),波兰哥白尼索引,中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:91748