Study On The Role Of PDK4 In Regulating The Lipid Metabolism Of Intramuscular Adipocytes In Goats

Pyruvate Dehydrogenase Kinase 4(PDK4)is a mitochondrial protein that encodes a histidine kinase domain.It plays an important role in many physiological processes,such as fat metabolism,glycolysis,tricarboxylic acid cycleand ATP formation.However,the role of PDK4 in the lipid metabolism of goats(Capra Hircus)has not been reported.Interestingly,the high-throughput sequencing results in our laboratory showed that PDK4 may be related to goat lipid metabolism.In this study,we cloned the gene sequence of goat PDK4 and performed its expression characteristics.Using the techniques of liposome transfection,real-time quantitative PCR(q PCR),overexpression and small interfering RNA(si RNA)to elucidate the effect of PDK4 on lipid metabolism in goat intramuscular preadipocytes.In addition,we used online prediction software to predicte its possible regulation mechanism.List the following research results:(1)Cloningthe sequence of goat PDK4 was 1808 bp,which containing the CDS 1224 bp.And this sequence encoded 407 amino acid residues.q PCR results showed that the expression level of PDK4 in lung,liver and arm triceps tissue were significantly higher than other tissues(P<0.01).(2)During the differentiation of goat intramuscular preadipocytes,the m RNA expression level of PDK4 was increased gradually and reached the peake at 5 d(P<0.01).(3)Inhibiting the expression of PDK4 was significantly reduced lipid droplet accumulation in goat intramuscular adipocytes.Furthermore,the m RNA expression level of Lipid absorption and transport genes CD36,fatty acid synthesis gene ACACA,triglyceride degradation genes HSL and ATGL,and triglyceride synthesis gene ADRP were significantly down-regulated(P<0.01).The m RNA expression level of lipid absorption and transport genes FABP3 and triglyceride synthesis gene AGPAT6 were significantly down-regulated(P<0.05).(4)Overexpression of PDK4 significantly increased lipid accumulation in goat intramuscular adipocytes.The expression level of fatty acid synthesis gene ACACA triglyceride degradation gene ATGL was significantly up-regulated(P<0.05).Moreover,lipid absorption and transport genes FABP3 and CD36,and triglyceride synthesis genes AGPAT6 and ADRP m RNA expression were significantly up-regulated(P<0.01).(5)Using KEGG pathway enrichment analysis we obtained results of PDK4 and mi RNAs that regulate PDK4.After getting the intersection,we found that PDK4 maight play a regulatory role via the signal pathways of AMPK,fatty acid biosynthesis,insulin,fatty acid metabolism,m TOR,ECM-receptor interactions and so on.Together,the sequence of goat PDK4 was cloned,the tissue expression profile and cell time-series expression profile of PDK4 was constructed.Interference /overexpression showed that PDK4 promoted the lipid accumulation in goat intramuscular adipocytes,and this effect may achieve by regulating the expression level of FABP3,CD36,ACACA,AGRAT6,ADRP and ATGL.In addition,PDK4 may regulate lipid metabolism in goat intramuscular adipocytes via AMPK,fatty acid biosynthesis,insulin,fatty acid metabolism,m TOR and ECM-receptor and other signaling pathways.