The Effect Of Bovine Recombinant Adiponectin On The Key Enzymes Of Fat Metabolism In Dairy Cows

Adiponectin (ADPN)is an adipocyte-specific secreted factor that plays a role in fat metabolism, energy homeostasis and insulin sensitivity is one of the most abundant expression protein in the adipose tissue. It is the important factors of regulatory network for body's lipid metabolism and blood glucose homeostasis. The study for ketosis and fatty liver in dairy cows in the metabolism of fat and gluconeogenesis to provide a new basis.In this study using of the fluorescent quantitative RT-PCR method and Western Blotting detection methods, and the experiments of the primary adipocyte and hepatocytes cultured in vitro researched the effect of bovine recombinant adiponectin on fat metabolism in the key enzyme HSL, ACC, CPT-Ⅰ, HADH and HMG-CS in the mRNA expression levels and protein expression levels.To reveal the cause of the fat mobilization mechanisms originated perinatal cows negative energy balance, and the control of cytokine levels provides a theoretical basis.The fluorescent quantitative RT-PCR was employed to determine the effect of the expressing level of ADPN and HSL mRNA in the experiments of primary adipocyte culture in vitro ,which were treated with different concentrations of recombinant ADPN (0,2.5,5,12.5,25,50 and 100μg/ml) for 24 hours. The results showed that adiponectin can suppress the autologous expression with negative feedback . The expression of HSL that is adiponectin dose-dependent manner reached highest with the medium supplemented with bovine recombinant adiponectin concentration 15μg/ml. At the same time with the concentration increasing the expression decreased.The fluorescent quantitative RT-PCR combining WB was employed to determine the effect of the expressing level of ACC,CPT-Ⅰ,HADH and HMG-CS mRNA and protein in the experiments of primary hepatocyte culture in vitro ,which were treated with different concentrations of recombinant ADPN (0,2.5,5,12.5,25,50 and 100μg/ml) for 24 hours. The results showed that adiponectin effect on the expression of ACC, HADH and HMG-CS ,but the effect is not obvious . In constrast adiponectin can promote the expression of CPT-Ⅰ, but high concentrations inhibit the expression of ADPN . And the expression of CPT-Ⅰcan reach highest with the ADPN concentration of 12.5μg / ml . ADPN promote the increase the fat metabolism mainly through increased the channel of fat mobilization and membrane transport. Western Blotting results show that: the test of adiponectin effect on the expression of the enzyme is mainly the transcriptional level, the level of translation has no immediate impact...