creb1对肝星状细胞tgfβ3表达及其启动子活性的影响word格式论文.docxVIP

独创性声明本人声明所呈交的学位论文是我个人在导师的指导下进行的研究工作及取得的研究成果。据我所知，除文中已标明引用的内容外，本论文不包含任何其他人或集体已经发表或撰写过的研究成果。对本文的研究做出贡献的个人和集体，均已在文中以明确方式标明。本人完全意识到本声明的法律结果由本人承担。学位论文作者签名：日期：年月日学位论文版权使用授权书本学位论文作者完全了解学校有关保留、使用学位论文的规定，即：学校有权保留并向国家有关部门或机构送交论文的复印件和电子版，允许论文被查阅和借阅。本人授权华中科技大学可以将本学位论文的全部或部分内容编入有关数据库进行检索，可以采用影印、缩印或扫描等复制手段保存和汇编本学位论文。本论文属于必威体育官网网址□，在年解密后适用本授权书。√ 不必威体育官网网址□。（请在以上方框内打“√”）学位论文作者签名：指导教师签名：日期：年月日日期：年月日目录中文摘要················································································1ABSTRACT2研究背景················································································4材料及方法··········································································· 4结果·················································································7讨论·················································································9参考文献·············································································12综述··················································································18致谢··················································································29附录：···············································································30CREB-1对肝星状细胞TGF-β3表达及其启动子活性的影响中文摘要目的探讨在大鼠肝星状细胞(HSC)中cAMP反应元件结合蛋白-1(CREB-1)对转化生长因子-β3(TGF-β3)mRNA表达及其启动子活性的影响。方法将HSC分为CREB-1表达质粒转染组(C)，siRNA-CREB-1转染组(S)，阴性对照质粒组(N)；Forskolin处理组(F)，H-89处理组(H)及空白组(B)。以上各组根据加或不加入外源性TGF-β3重组蛋白再分为(+)组和(-)组，运用实时荧光定量PCR法检测TGF-β3mRNA的表达。上述各组共转染PGL3-basic-TGF-β3P(W)或PGL3-basic-TGF-β3P-mCRE(M)，以pRL-SV40为空白，荧光素酶报告基因分析法检测各组TGF-β3启动子活性。结果TGF-β3mRNA表达结果显示，S(-)组较N(-)组明显降低(P=0.021)，C(-)组较N(-)组表达有升高趋势(P=0.082)；S(+)组较N(+)组明显降低(P=0.007)，C(+)组较N(+)组表达有升高趋势(P=0.096)。H(-)组与B(-)组，H(+)组与B(+)，N(+)组与N(-)组，S(+)组与S(-)组，B(+)组与B(-)组，H(+)组与H(-)组分别比较，差异具有统计学意义(P值分别为0.011,0.04,0.018,0.025,0.047,0.013)。荧光素酶报告基因分析结果显示，S(W)组、N(W)组、C(W)组TGF-β3启动子活性依次升高(P值分别为0.004,0.019)；S(M)组、N(M) 组、C(M)组TGF-β3启动子活性差异无统计学意义(P值分别为0.680,0.935)。H(W)组、B(W)组、F(W)组TGF-β3启动子活性依次升高(P值分别为0.036,0.012)；H(M)组、B(M)组、F(M)组TGF-β3启动子活性差异无统计学意义(P值分别为0.097,0.592)。结论增加HSC内CREB-1表