【作者】 陈潇；

【导师】 谢红付；

【作者基本信息】 中南大学 ， 皮肤病与性病学， 2008， 硕士

【摘要】 目的观察5-氮杂胞苷(5-azaC)对人角质形成细胞增殖及诱导型一氧化氮合酶(iNOS)表达的影响,并初步探讨甲基化状态与iNOS表达的关系。方法原代培养人角质形成细胞,用浓度为1μM的5-azaC处理细胞24h,72h和120h,对照组细胞以PBS处理。通过MTT实验观察细胞增殖活性的差异,RT-PCR及Western Blot分别检测iNOS mRNA和蛋白的表达情况。结果1.由MTT法绘制细胞生长曲线,1μM 5-azaC处理后人角质形成细胞与对照组相比生长速度减慢;2.所有对照组及1μM 5-azaC处理后24h人角质形成细胞均未检出iNOS mRNA/蛋白表达;1μM 5-azaC处理后72h和120h人角质形成细胞有iNOS mRNA/蛋白表达,且120h表达量强于72h(P＜0.05)结论5-azaC可能通过降低人角质形成细胞iNOS基因甲基化水平,从而导致其表达,进而抑制细胞增殖。更多还原

【Abstract】 ObjectiveTo observe the effects of 5-azaC on proliferation and inducible nitric oxide synthase expression in normal human keretinocytes,and then explore the relation between methylation state and iNOS expression.MethodsPrimary normal keratinocytes were cultured and treated with 1μM 5-azaC for 24,72 and 120h,and the control group was treated with PBS. The activation difference in cell growth was observed by MTT assay, and the iNOS mRNA and protein expression of keratinocytes on different time were measured by RT- PCR and Western blot,respectively.Results1.In the graph of cell growth by MTT assay,the keratinocytes treated with 1μM 5-azaC showed a slower growth rate in contrast to the control group.2.The iNOS mRNA and protein of keratinocytes could not be detected on control groups and cells treated with 1μM 5-azaC at 24h;the cells were treated with 1μM 5-azaC and iNOS mRNA and protein were detected at 72h and 120h,and the expression of iNOS was more significantly increased at 120h than 72h(P＜0.05).Conclusion The 5-azaC can induce the iNOS expression of normal human keretinocytes,it maybe take effect on degrading the methylation state of iNOS gene,and then restrain the proliferation of normal human keretinocytes.更多还原