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Background and purpose of the establishment of the C57BL / 6 mouse hepatitis B virus HBV infection in an animal model of chemotactic factor IL-18, Mig, IP-10, CXCL12, CCR7, CCL19 and some acute reference gene chip results, detection model liver tissue phase protein genes, immune-related protein gene expression differences, and to lay the foundation for further study of inflammatory factors in effect and mechanism of HBV infection. Experimental Method 1. Establish animal model of C57BL / 6 mice of HBV infection: pBluescript-HBV plasmid transformed the ultracompetent bacterial DH5α was extracted large plasmid, the high-pressure hydrodynamic method in the mouse tail vein injection, the establishment of infection model, while establishing the no-load control group (pBluescript) animal models. 2. Modeling Day 4 the fetching model animal peripheral blood ELISA kit to detect the expression levels of peripheral blood HBsAg, validation infection model whether successfully established receive liver tissue cryopreservation. 3 refer to the detection result of gene chip, the selection of interested differentially expressed chemotactic factor, primers were designed. Trizol extraction of total RNA from liver tissue, reverse transcribed into cDNA, Real-time PCR detection of the experimental and control groups corresponding to the number of copies of the gene factor analysis of the RT-PCR gel electrophoresis. 4 immunohistochemical method for the expression of the difference is more obvious factor IP-10 for detection. Results 1.HBV infected C57BL / 6 mouse model: ELISA detects the tail vein of high-pressure injection of HBV C57BL / 6 mice group HBsAg results were positive, show that the high-pressure injection of HBV infection through the tail vein of C57BL / 6 mouse model successfully established. The 2.HBV infection of C57BL / 6 mouse model of liver tissue expression of chemokine the: ① Real-time PCR test results show that the amplification curve and melting curve, the experimental group compared with the control group without load, IL-18, Mig IP-10, Cxcl12, Ccl19, Ccr7 of expression were increased in various degrees to IP-10, the most significant; RT-PCR amplification product was gel electrophoresis visible specific primers amplified band, IP-10 corresponding article with the most significantly, the results showed that the above chemokine gene copy number in the experimental group increased to different degrees, and Real-time PCR test results showed a consistent trend; the ② immunohistochemistry chemical test results show that the experimental group IP-10 expression. The 3.HBV infection of C57BL / 6 mouse model of liver tissue molecules of the immune response and acute protein molecule expression: Real-time PCR results showed that the amplification and dissolution curves, compared with the control group, the acute phase response in the experimental group the molecular Saa2 increased expression; RT-PCR amplification The product of electrophoresis comparison with the control group, the experimental group of heat shock protein gene Dnajc10, the acute phase response related factors Saa2, the innate immune response molecules Masp1, as well as the expression of anti-apoptotic factor Vnn1 There are varying degrees. Conclusions based on the results of ELISA for detection of HBsAg of HBV infection of C57BL / 6 mice animal model of success. Real-time PCR and RT-PCR showed that the chemokine IL-18, Mig, IP-10, Cxcl12 Ccl19 Ccr7 increased in the experimental group; PCR results with the results of immunohistochemistry showed that IP-10 factor expression increased significantly, suggesting that HBV infection chemokine expression increased in varying degrees, as well as the expression of immune response-related molecules have different levels of expression, its expression increased role and its mechanism needs further study.
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