The Study Of Mouse Adiponectin's Protective Role In Con A-induced Liver Injury

Adiponectin, an adipocytokine, is produced exclusively by the adipocyte. Besides modulating the metabolism of fat and glucose and enhancing the insulin sensitivity, it exhibits ability to regulate immunity and inflammatory reactions by inhibiting the production and functions of tumor necrosis factor-alpha (TNF-α). As the TNF-α-induced hepatocytes immunologic injury, mediated by T cell activation, is the main mechanism underlying the viral hepatitis. Therefore, the Con A-induced hepatitis in mice was used to evaluate the role of adiponectin in regulating TNF-a production and fulminant autoim muneologic liver damage following the concanavalin A injection.The thesis is composed by four parts: (1) Cloning of mouse adiponectin gene (mAd) and expression of globular domain of adiponectin in E coli; (2) Construction of recombinant plasmid containing mouse adiponectin gene, named 'pAA-neo-mAd', and verification of the expression of recombinant vector in COS-7 cell; (3) Delivering two recombinant plasmids, pAA-neo-mAd and pCI-neo-mAd, into mice body by hydrodynamics-based procedure, and comparing the expression efficiency of two vectors; (4) Based on the expression of vector-derived adiponectin (3 days after recombinant plasmids pAA-neo-mAd injected into mice by tail vein), Con A was administered into the mice, using the mice transfected with empty pAA-neo vector as control. Then, the serum levels of alanine aminotransferase (ALT), TNF-α and adiponectin were detected, and livers histological examinations were carried out on different time points after Con A injection.Through statistical analysis, the results show that adiponectin is effective in protecting hepatocytes from TNF-α-induced immunological injury. The mechanism of protective effect is mainly by inhibiting the synthesis and/or release of TNF-α.The present study also suggests that adiponectin may be a useful therapeutic tool for viral hepatitis in future.