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缺氧诱导体外培养人视网膜色素上皮细胞中Toll样受体4的表达
  • ISSN号:1672-5123
  • 期刊名称:《国际眼科杂志》
  • 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]第四军医大学西京医院眼科全军眼科研究所,中国陕西省西安市,710032
  • 相关基金:基金项目:中国国家自然科学基金资助项目(No.30371516,30672291)
中文摘要:

目的:了解缺氧对人视网膜色素上皮(retinal pigment epithelium, RPE)细胞Toll样受体4(Toll-like receptor 4,TLR4)表达的影响.方法:采用200μmol/L CoCl2处理RPE细胞建立化学缺氧模型,以未处理细胞作对照,在缺氧处理后2, 4, 8, 12和24h用免疫荧光法观察TLR4在细胞中的定位,用RT-PCR和Western blot检测细胞中TLR4表达水平.结果:共聚焦显微镜观察到正常对照组RPE细胞胞质内有微弱的TLR4荧光表达,缺氧后胞质内荧光表达明显增强,缺氧12h荧光强度达最强.RT-PCR和Western blot结果分别提示随缺氧时间延长,RPE细胞中TLR4 mRNA和蛋白表达增强水平升高,缺氧至12h表达高峰最强(P〈0.05),24h表达开始下降,各组差异有统计学意义.结论:缺氧可诱导RPE细胞中TLR4表达增强.

英文摘要:

AIM: To observe the effects of hypoxia on the expression of Toll-like receptor 4 (TLR4) in cultured human retinal pigment epithelium (RPE) cells. METHODS : Chemical hypoxic model was established by treating the cultured human RPE cells with 200μmol/L CoCI2. The location of TLR4 in human RPE cells was observed by immunofluorescence assay, and RT-PCR together with Western blot were applied to detect the expression of TLR4 2, 4, 8, 12, 24 hours after the treatment. RESULTS: Immunofluorescence assay revealed that TLR4 was slightly expressed in human RPE cells under normoxic condition. After hypoxic treatment, there was a significant increase in green fluorescence within the cytoplasm. TLR4 expression tend to intensify over time and peaked at 12 hours, which was confirmed by RT-PCR and Western blot assay exhibiting TLR4 mRNA/protein levels ( P 〈 0.05). The expression levels began to decrease at 24 hours, and there was statistical significance among the groups. CONCLUSION : Hypoxia can strengthen TLR4 expression in cultured human RPE cells.

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期刊信息
  • 《国际眼科杂志》
  • 中国科技核心期刊
  • 主管单位:中国陕西省卫生和计划生育委员会
  • 主办单位:中华医学会西安分会
  • 主编:惠延年
  • 地址:中国西安友谊东路269号
  • 邮编:710054
  • 邮箱:IJO2000@126.com;IJO.2000@163.com
  • 电话:029-85263940
  • 国际标准刊号:ISSN:1672-5123
  • 国内统一刊号:ISSN:61-1419/R
  • 邮发代号:52-239
  • 获奖情况:
  • 2000年被美国中华医学会评为优秀期刊
  • 国内外数据库收录:
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  • 被引量:22343