Study on the Regulation of Leptin-mediated JAK/STAT Signal Pathway on Lipid Metabolism in Porcine Subcutaneous Preadipocyte

doi:10.13560/j.cnki.biotech.bull.1985.2020-0749

Abstract

Abstract:

Using in vitro cultured porcine subcutaneous preadipocytes as the study material,the expression levels of related genes were measured for investigating the molecular regulation mechanism of Leptin-mediate JAK/STAT signal pathway on lipid metabolism. The subcutaneous adipocytes were treated with 0 and 100 ng/mL Leptin for 48 h,and the dipocytes were identified by oil red O dyeing,and the contents of triglyceride and free fatty acids in the cells were measured by kit,and the mRNA expression levels of relate genes in the JAK/STAT signal pathway of subcutaneous preadipocytes were detected using real-time PCR technology. The results showed the content of the triglyceide of the subcutaneous preadipocyte that in Leptin group was significantly lower（P<0.05）than that in the control group,and the content of the free fatty acids was lower than that of the control group. The expression levels of gene lepR,JAK2,STAT3,CPT-1,ACOX1,and PGC-1 in JAK/STAT signal pathway in Leptin group were significantly higher than that in control group（P<0.05）.The expression levels of gene lepR,JAK2,STAT3,CPT-1,ACOX1,and PGC-1 were significantly negatively correlated with the content of triglyceride and free fatty acid（P<0.05）. In conclusion,the results suggested that Leptin activated JAK/STAT signal pathway in subcutaneous preadipocyte,increased the mRNA expressions of relevant genes in the JAK/STAT signaling pathway,promoted the oxidation of fatty acid and the decomposition of triglyceride,and thus decreased the content of triglycerides and free fatty acids in subcutaneous preadipocyte.

Key words: subcutaneous preadipocyte, Leptin, JAK/STAT, signal pathway, gene expression

ZHA Xing-qin, YANG Ming-hua, LI Yong-neng, ZHAO Su-mei, HUANG Ying. Study on the Regulation of Leptin-mediated JAK/STAT Signal Pathway on Lipid Metabolism in Porcine Subcutaneous Preadipocyte[J]. Biotechnology Bulletin, 2021, 37(2): 88-95.

Figures/Tables 6

References 16

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目的基因	引物序列（5'-3'）	退火温度/℃	产物大小/bp	GenBank登录号
18S rRNA	F：TAGAGTCACCAAAGCCGC	55	194	NR_002170.3
18S rRNA	R：TCAGTTATGGTTCCTTTG	55	194	NR_002170.3
LepR	F：GTGATAACTGCATTTGACTTGGC	62	271	NM001024587
LepR	R：CTGCAATGTTGTCTGCATGTACAG	62	271	NM001024587
JAK2	F：ATTTGAAGACCGAGACCC	60	272	NM_214113.1
JAK2	R：TTAGATTACGCCGACCAG	60	272	NM_214113.1
STAT3	F：CAGTAAGGAGAGGGAGCG	56	254	NM_001044580
STAT3	R：AGAGGTAGACCAGCGGAG	56	254	NM_001044580
CPT-1	F：ATGGTGGGCGACTAACT	60	321	AY181062
CPT-1	R：TGCCTGCTGTCTGTGAG	60	321	AY181062
ACOX1	F：CGGGCACGGCTATTCTCAC	56	193	NM_001101028
ACOX1	R：CTGGCTGGGCAGGTCATTC	56	193	NM_001101028
PGC-1α	F：CCCAGTCTGCGGCTATTTG	56	282	AB106108.1
PGC-1α	R：GGGATGACCGAAGTGCTTG	56	282	AB106108.1

目的基因	引物序列（5'-3'）	退火温度/℃	产物大小/bp	GenBank登录号
18S rRNA	F：TAGAGTCACCAAAGCCGC	55	194	NR_002170.3
18S rRNA	R：TCAGTTATGGTTCCTTTG	55	194	NR_002170.3
LepR	F：GTGATAACTGCATTTGACTTGGC	62	271	NM001024587
LepR	R：CTGCAATGTTGTCTGCATGTACAG	62	271	NM001024587
JAK2	F：ATTTGAAGACCGAGACCC	60	272	NM_214113.1
JAK2	R：TTAGATTACGCCGACCAG	60	272	NM_214113.1
STAT3	F：CAGTAAGGAGAGGGAGCG	56	254	NM_001044580
STAT3	R：AGAGGTAGACCAGCGGAG	56	254	NM_001044580
CPT-1	F：ATGGTGGGCGACTAACT	60	321	AY181062
CPT-1	R：TGCCTGCTGTCTGTGAG	60	321	AY181062
ACOX1	F：CGGGCACGGCTATTCTCAC	56	193	NM_001101028
ACOX1	R：CTGGCTGGGCAGGTCATTC	56	193	NM_001101028
PGC-1α	F：CCCAGTCTGCGGCTATTTG	56	282	AB106108.1
PGC-1α	R：GGGATGACCGAAGTGCTTG	56	282	AB106108.1

指标	对照组	Leptin组
甘油三酯（mmol/L）	0.15±0.02^a	0.07±0.01^b
游离脂肪酸（μmol/gprot）	1.16±0.13^a	0.94±0.09^a

指标	对照组	Leptin组
甘油三酯（mmol/L）	0.15±0.02^a	0.07±0.01^b
游离脂肪酸（μmol/gprot）	1.16±0.13^a	0.94±0.09^a