The Effects Of JAZF1Gene Inhibits In AKT And AMPK Signaling Pathway On Giucose And Lipid Metabolism

Objective: To establish the mice JAZF1-shRNA expression vectorand construct the cell model of JAZF1gene inhibition.Methods: JAZF1shRNA sequence were designed and synthesized.Select pGenesil-1.2carrier and mU6promoter to build pGenesil-JAZF11,2,3inhibit the plasmid. Transfected into the mice hepa1-6cells,quantitative PCR technique were used to screen the best inhibition ofplasmid. The effect was determined in3T3-l1cells.Result: The three recombinant plasmids of pGenesil-JAZF1wereconstructed successfully. The pGenesil-JAZF1-3inhibition of plasmid wasscreened for the best suppression fragment,and inhibition rate is78-84%inhepa1-6cell and37-42%in3T3-L1cells.Conclusion: JAZF1-shRNA recombinant plasmid Was successfullyconstructed, and the best efficiency was identified. Objective: To observe the effect of JAZF1suppression onglycolipids metabolism of adipocyte and hepatocytes.Method：3T3cells and Hepa1-6cells were transfected with mostefficiency pGenesil-JAZF1-3plasmids, cell immunofluorescence wereemployed to detect glycometabolism related protein expression, Westernblot and cell immunofluorescence were used to detect glycolipidsmetabolism related protein level.Glycometabolism targets were detectedusing real-time PCR,Triglyceride level of3T3cells after transfection wasassessed by oil red O stain，H3tracing was used to assess carbohydrateuptake rate.Results： GLUT1and GLUT4mRNA was upregulated afterpGenesil-JAZF1-3transfection（P<0.05），protein level of GLUT1andG-6-Pase were upregulated but didn’t reach statistics significance, PEPCKwas upregulated （P<0.05）；HSL and PPAR-γ was upregulated but didn’treach statistics significance。In3T3cells, GLUT1and GLUT4mRNA wasdownregulated after pGenesil-JAZF1-3transfection（P<0.05），GLUT1and GLUT4mRNA was upregulated but didn’t reach statistics significance;pGenesil-JAZF1-3transfected showed lower expression of HSL(P<0.05）and higher expression of PPAR-γ;Visfatin、Acrp30and Insig-2weredownregulated after pGenesil-JAZF1-3transfection（P<0.05）. Conclusion： JAZF1gene silence reduced basic carbohydratetransport，raised synthesis of lipide and cholesterol, cytokine related to fatdesintegration was downregulated at the same time. Objective：To study on the signal pathway that related to theglycolipids metabolism, identify effect of JAZF1suppresion onglycolipids metabolism.Methods：Protein targets on signal pathway that related to theglycolipids metabolism were assesed using Western blot technology.Results：P-AKT and P-AMPK of Hepa1-6after JAZF1silence wasno changed. AKT/PKA/HSL lipid metabolism signal and AMPK/TORC2glycometabolism signal were inhibited after JAZF1silence in3T3cells（P<0.01）.Conclusion：The inhibition of JAZF1gene was a important factor insignal pathway of glycolipids metabolism,which increase the risk ofdiabetes. JAZF1gene may promote glucose and lipid metabolismsignaling pathway in the phosphorylation as an important factors.