重组9型腺相关病毒转染大鼠心脏成纤维细胞及对nf kb通路的影响-recombinant adeno-associated virus type 9 transfected rat cardiac fibroblasts and its effect on nf kb pathway.docxVIP

下载本文档

7
0
约3.24万字
约 34页
2018-09-08 发布于上海
举报
版权申诉

重组9型腺相关病毒转染大鼠心脏成纤维细胞及对nf kb通路的影响-recombinant adeno-associated virus type 9 transfected rat cardiac fibroblasts and its effect on nf kb pathway.docx

1、本文档共34页，可阅读全部内容。
2、原创力文档（book118）网站文档一经付费（服务费），不意味着购买了该文档的版权，仅供个人/单位学习、研究之用，不得用于商业用途，未经授权，严禁复制、发行、汇编、翻译或者网络传播等，侵权必究。
3、本站所有内容均由合作方或网友上传，本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺！文档内容仅供研究参考，付费前请自行鉴别。如您付费，意味着您自己接受本站规则且自行承担风险，本站不退款、不进行额外附加服务；查看《如何避免下载的几个坑》。如果您已付费下载过本站文档，您可以点击这里二次下载。
4、如文档侵犯商业秘密、侵犯著作权、侵犯人身权等，请点击“版权申诉”（推荐），也可以打举报电话：400-050-0827(电话支持时间：9:00-18:30)。
5、该文档为VIP文档，如果想要下载，成为VIP会员后，下载免费。
6、成为VIP后，下载本文档将扣除1次下载权益。下载后，不支持退款、换文档。如有疑问请联系我们。
7、成为VIP后，您将拥有八大权益，权益包括：VIP文档下载权益、阅读免打扰、文档格式转换、高级专利检索、专属身份标志、高级客服、多端互通、版权登记。
8、VIP文档为合作方或网友上传，每下载1次，网站将根据用户上传文档的质量评分、类型等，对文档贡献者给予高额补贴、流量扶持。如果你也想贡献VIP文档。上传文档

重组9型腺相关病毒转染大鼠心脏成纤维细胞及对nf kb通路的影响-recombinant adeno-associated virus type 9 transfected rat cardiac fibroblasts and its effect on nf kb pathway

新疆医科大学医学硕士学位论文新疆医科大学医学硕士学位论文 Abstract Abstract PAGE PAGE 2 PAGE PAGE 11 的表达受到抑制。结论：AAV9-EGFP 能够有效转染大鼠心脏成纤维细胞，转染后 EGFP 基因能够有效得以表达， rAAV9 对细胞增殖无明显抑制。应用 rAAV9-EGFP-R65 能够抑制大鼠心脏成纤维细胞 NF-κB 活性，并能够抑制大鼠心脏成纤维细胞 P65 蛋白表达。关键词：重组 9 型腺相关病毒；抗 NF-κB 核酶基因；大鼠心脏成纤维细胞；转染；核转录因子-κB rAAV9 transfection of rat cardiac fibroblasts cells in vitro and effect of Nuclear Factor-κB activity Postgraduate: Du Lei Supervisor: Prof. Ma Yi-tong Abstract Objective: To assess the transfection efficiency of recombinant adeno-associated virus serotype 9 mediated enhanced green fluorescent protein (rAAV9-EGFP) to rat cardiac fibroblasts and the impact on growth of rat cardiac fibroblasts. And to evaluate the transfection efficiency using recombinant adeno-associated virus Serotype 9 mediated Anti-NF-κB ribozyme and enhanced green fluorescent protein (rAAV9 -EGFP-R65) to cardiac fibroblasts cells and on the effect of Nuclear Factor-κB (NF-κB) activity. Methods: The rat cardiac fibroblasts were collected and cultured. The rAAV9 carrying enhanced green fluorescence protein (rAAV9-EGFP) gene was transfected into rat cardiac fibroblasts at different multiplicities of infection (MOI=1×104, 1×105, 1×106). EGFP expression in the cells was observed under inverted fluorescence microscope, and the EGFP-positive cell percentage was determined by flow cytometry. AlamarBlue assay was used to assess the proliferation of the transfected cells. The quantitative analysis of the DNA binding activity of NF-κB was examined by electrophoretic mobility shift assay (EMSA). The cells were divided into controlled group, rAAV9-EGFP-R65 tranfected group, TNF-α treated group and TNF-α treated rAAV9-EGFP-R65 tranfected group. Results: The cells with rAAV9-EGFP transfection at MOI of 1×106 began to exhibit EGFP expression 24 hours after transfection and the cells transfection at MOI of 1×105 and 1×104 began to exhibit EGFP expression 48 hours after transfection. The cells growth was normal in all three groups under