Effect of PDK4 on the Lipid Metabolism of Goat Intramuscular Adipocytes

doi:10.13560/j.cnki.biotech.bull.1985.2021-0368

Abstract

Abstract:

This study aims to clone the goat PDK4 gene sequence,to clarify its tissue and temporal expression characteristics,and to clarify its effect on the lipid metabolism of goat intramuscular adipocytes. Overexpression and interference cell model of goat PDK4 was constructed,and RT-PCR and real-time fluorescent quantitative PCR were used to determine and study the effect of PDK4 on the lipid metabolism in goat intramuscular adipocytes. As results,the cloned goat PDK4 sequence was 1 808 bp,and it was located in mitochondria and cytoplasm. The expression pattern of goat PDK4 tissue and cell temporal expression was clarified,showing that PDK4 was highly expressed in the goat’s lung,triceps brachii muscle and liver tissue（P<0.01）. The expression level in goat intramuscular adipocytes at 5 d of induction was significantly higher than that before differentiation（P<0.01）. Interference and overexpression of goat PDK4 significantly reduced and increased lipid accumulation,respectively. The expression of lipid metabolism-related genes FABP3,CD36,ACACA,AGPAT6 and ADRP significantly（P<0.05）or extremely significantly（P<0.01）decreased with si-PDK4,while those genes extremely significantly increased（P<0.01）after overexpression of PDK4. In conclusion,PDK4 is expressed in goat tissues and in various stages of intramuscular preadipocyte differentiation. Overexpression of goat PDK4 promots the up-regulation of lipid droplet accumulation and lipid deposition-related gene expression in adipocytes,and shows the opposite trend and expression after interference.

Key words: pyruvate dehydrogenase kinase 4, goat, expression characteristics, lipid metabolism

ZHANG Hao, ZHANG Ya-nan, LI Xin, WANG Jia-mei, WANG Yong, ZHU Jiang-jiang, XIONG Yan, LIN Ya-qiu. Effect of PDK4 on the Lipid Metabolism of Goat Intramuscular Adipocytes[J]. Biotechnology Bulletin, 2021, 37(12): 151-159.

Figures/Tables 7

References 36

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基因 Gene	登录号 GenBank	退火温度 Annealing temperature/℃	序列 Primer sequence（5'-3'）	产物长度 Product length/bp	用途 Purpose
PDK4	NM_001101883.1	60	S:GAGTCTCAAGCCACCTACGG	1 806	PCR
PDK4	NM_001101883.1	60	A:TGACATCAATCAGTGGCCTAA	1 806	PCR
PDK4	MF564045.1	53	S:ACCTCTTTTGCATTTTTGCG	111	qPCR
PDK4	MF564045.1	53	A:TTTGGTCCTCTGGGCTTTTC	111	qPCR
TBP	XM_018053502.1	60	S:AACAGCCTCCCACCTTATGC	155	qPCR
TBP	XM_018053502.1	60	A:TGCTGCTCCTCCAAAATAGAC	155	qPCR
CD36	X91503	60	S:GTACAGATGCAGCCTCATTTCC	81	qPCR
CD36	X91503	60	A:TGGACCTGCAAATATCAGAGGA	81	qPCR
FABP3	NM_174313.2	58	S:GATGAGACCACGGCAGATG	120	qPCR
FABP3	NM_174313.2	58	A:GTCAACTATTTCCCGCACAAG	120	qPCR
AGPAT6	JI861797.1 XM_018041984.1	60	S:AAGCAAGTTGCCCATCCTCA	101	qPCR
AGPAT6	JI861797.1 XM_018041984.1	60	A:AAACTGTGGCTCCAATTTCGA	101	qPCR
ADRP	HQ846827 NM_001285596.1	60	S:TACGATGATACAGATGAATCCCAC	202	qPCR
ADRP	HQ846827 NM_001285596.1	60	A:CAGCATTGCGAAGCACAGAGT	202	qPCR
ATGL	GQ918145	60	S:GGAGCTTATCCAGGCCAATG	180	qPCR
ATGL	GQ918145	60	A:TGCGGGCAGATGTCACTCT	180	qPCR
UXT	NM_001037471	60	S:CAGCTGGCCAAATACCTTCAA	125	qPCR
UXT	NM_001037471	60	A:GTGTCTGGGACCACTGTGTCAA	125	qPCR

基因 Gene	登录号 GenBank	退火温度 Annealing temperature/℃	序列 Primer sequence（5'-3'）	产物长度 Product length/bp	用途 Purpose
PDK4	NM_001101883.1	60	S:GAGTCTCAAGCCACCTACGG	1 806	PCR
PDK4	NM_001101883.1	60	A:TGACATCAATCAGTGGCCTAA	1 806	PCR
PDK4	MF564045.1	53	S:ACCTCTTTTGCATTTTTGCG	111	qPCR
PDK4	MF564045.1	53	A:TTTGGTCCTCTGGGCTTTTC	111	qPCR
TBP	XM_018053502.1	60	S:AACAGCCTCCCACCTTATGC	155	qPCR
TBP	XM_018053502.1	60	A:TGCTGCTCCTCCAAAATAGAC	155	qPCR
CD36	X91503	60	S:GTACAGATGCAGCCTCATTTCC	81	qPCR
CD36	X91503	60	A:TGGACCTGCAAATATCAGAGGA	81	qPCR
FABP3	NM_174313.2	58	S:GATGAGACCACGGCAGATG	120	qPCR
FABP3	NM_174313.2	58	A:GTCAACTATTTCCCGCACAAG	120	qPCR
AGPAT6	JI861797.1 XM_018041984.1	60	S:AAGCAAGTTGCCCATCCTCA	101	qPCR
AGPAT6	JI861797.1 XM_018041984.1	60	A:AAACTGTGGCTCCAATTTCGA	101	qPCR
ADRP	HQ846827 NM_001285596.1	60	S:TACGATGATACAGATGAATCCCAC	202	qPCR
ADRP	HQ846827 NM_001285596.1	60	A:CAGCATTGCGAAGCACAGAGT	202	qPCR
ATGL	GQ918145	60	S:GGAGCTTATCCAGGCCAATG	180	qPCR
ATGL	GQ918145	60	A:TGCGGGCAGATGTCACTCT	180	qPCR
UXT	NM_001037471	60	S:CAGCTGGCCAAATACCTTCAA	125	qPCR
UXT	NM_001037471	60	A:GTGTCTGGGACCACTGTGTCAA	125	qPCR