c57bl6小鼠乙型肝炎病毒感染模型中细胞因子研究word格式论文.docxVIP

检测结果表明，扩增曲线和溶解曲线良好，实验组与空载对照组相比，IL-18，Mig，IP-10，Cxcl12，Ccl19，Ccr7 的表达均有不同程度的增高，以 IP-10 最为显 著；RT-PCR 扩增产物凝胶电泳可见特异引物扩增的条带，IP-10 相应条带最为明 显，结果表明实验组中上述趋化因子基因拷贝数有不同程度增加，与 Real-time PCR 检测结果呈一致趋势；②免疫组织化学方法检测结果表明，实验组 IP-10 表 达增加。3．HBV 感染 C57BL/6 小鼠模型肝组织免疫应答分子和急性蛋白分子的表 达：Real-time PCR 结果表明，扩增曲线和溶解曲线良好，与对照组相比，实验 组中急性期应答相关分子 Saa2 表达增高；RT-PCR 扩增产物电泳图与对照组比 较，实验组中热休克蛋白相关基因 Dnajc10、急性期应答相关因子 Saa2、固有免 疫应答分子 Masp1、以及抗凋亡因子 Vnn1 的表达均有不同程度增加。结 论依据 ELISA 检测 HBsAg 结果表明，HBV 感染 C57BL/6 小鼠动物模型建立 成功。Real-time PCR 和 RT-PCR 均显示趋化因子 IL-18、Mig、IP-10、Cxcl12、 Ccl19、Ccr7 在实验组有不同程度的增高；PCR 结果与免疫组织化学检测结果均 表明 IP-10 因子表达增加明显，提示 HBV 感染中趋化因子的表达不同程度增加， 以及免疫应答相关分子的表达均有不同程度的表达增加，其表达增高作用及其机 制尚待进一步研究。关键词：乙型肝炎病毒；炎性因子；C57BL/6 小鼠动物模型Cytokines in C57BL/6 Mice Model of Hepatitis B Virus InfectionABSTRACTBackgrounds and ObjectiveBy establishing the C57BL/6 mice model of HBV infection and considering the gene microarrays as the references, the differences in the expression of chemokines such as IL-18，Mig，IP-10，CXCL12，CCR7 and CCL19, as well as the partial genes for acute phase proteins and immune-associated proteins were detected to further study the function and mechanism of the inflammatory factors in HBV infection.MethodsThe establishment of C57BL/6 mice model of HBV infection: with the preparation of super-competent bacteria DH5α, pBluescript and pBluescript-HBV plasmids were extracted to the mice by the method of high pressure water via caudal vein injection. And the mice with pBluescript and pBluescript-HBV were assigned into control and experimental group respectively.In both groups the peripheral blood was sampled to test the levels of HBsAg at 4th day. And the tests were estimated the establishment of the animal model. Meanwhile the livers in both groups were cryopreserved.According to the specific cytokines expressed significantly different, the primers were designed and sequences were synthesized. By method of Trizol, the hepatic total RNA was extracted to synthesize the primers. The numbers of the objecti