单细胞凝胶电泳检测慢性乙型肝炎肝癌患者的DNA损伤.docVIP

单细胞凝胶电泳检测慢性乙型肝炎肝癌患者的DNA损伤 摘 要 为探讨乙型肝炎病毒引起DNA损伤与肝癌形成的关系，以及丹参酮对DNA的保护作用，本课题使用单细胞凝胶电泳(SCGE)技术以及IMI 1.0彗星分析软件在体外观察健康组、慢性乙型肝炎组、慢性乙型肝炎合并肝癌组及慢性乙型肝炎组经丹参酮、H2O2处理后外周血淋巴细胞DNA的彗星状荧光图像。观察指标为彗星状荧光图像的彗星尾长、尾%DNA、尾惯量、Olive尾矩。 结果显示：健康组经SCGE技术后所得彗星图像未见明显彗星尾。慢性乙型肝炎组经SCGE技术后所得彗星图像可见有少量彗星尾。慢性乙型肝炎合并肝癌组经SCGE技术后所得彗星图像可见明显的彗星尾。慢性乙型肝炎合并肝癌组外周血淋巴细胞DNA形成的彗星尾长、尾%DNA、尾惯量和Olive尾矩4个指标均显著高于健康组和慢性乙型肝炎组（P0.05）。慢性乙型肝炎组各观察指标虽高于健康组，但无显著差异（P0.05）。丹参酮处理组与未处理组相比较无差异（P0.05）。H2O2处理组与未处理组相比较有显著差异（P0.05）。慢性乙型肝炎组经丹参酮及H2O2先后处理后发现各观察指标值减小，与单纯H2O2处理组比较有显著差异（P0.05）。 结论：慢性乙型肝炎合并肝癌患者外周血淋巴细胞DNA损伤明显。慢性乙型肝炎组外周血淋巴细胞DNA损伤与健康组比较无显著区别。加入丹参酮后可减少H2O2对慢性乙型肝炎患者外周血淋巴细胞DNA的损伤，证实丹参酮具有抗DNA氧化损伤的能力,对DNA有保护作用。 关键词： 单细胞凝胶电泳；彗星实验；乙型肝炎病毒；肝癌； 脱氧核糖核酸；丹参酮 A Study of DNA Damage in the Patients with Chronic Hepatitis B Complicated by HCC Using Single Cell Gel Electrophoresis Master Candidate: HE Wei Supervisor: TANG Yong-huang, Professor ABSTRACT Objective：In order to understand the relationship between DNA damage and hepatocellular carcinoma (HCC), and the tanshinone (丹参酮TAN) protection for this damage. Methods：We detected DNA in peripheral blood lymphocytes of three groups in healthy people, in patients with chronic hepatitis B and chronic hepatitis B complicated by HCC. In the group of patients with chronic hepatitis B we did treatment of both TAN and H2O2 in vitro. The tail extent, tail inertia, percentage of tail DNA and Olive tail moment are selected as parameters, which could be measured by the fluorescent pictures of single cell gel electrophoresis (SCGE) and IMI 1.0 comet assay analysis software. Results： We found that there were no obvious comet tail in healthy people and a little of comet tail in the group of patients with chronic hepatitis B. However, there was obvious comet tail in the group of patients with chronic hepatitis B complicated by HCC. The mean levels and percentage of these four parameters were significantly higher in the group of patients with chronic hepatitis B complicated by HCC than those both in healthy people