NR1H3基因调控猪前体脂肪细胞分化的研究

doi:10.11843/j.issn.0366-6964.2022.07.007

Abstract

Abstract: The purpose of this study was to investigate the developmental expression pattern of NR1H3 gene in subcutaneous adipose tissues of pigs at different ages and the effects of NR1H3 gene on adipogenic differentiation of porcine preadipocytes, and determine its primary function in fat deposition. qRT-PCR was used to detect the expression of NR1H3 in subcutaneous adipose tissues of Mashen pigs aged 30, 90 and 240 days. Porcine preadipocytes were isolated from dorsal subcutaneous adipose tissues of 5-day-old DLY (Duroc×(Landrace×Yorksire)) piglets. Cellular immunofluorescence technique was used to detect the Adiponectin protein content in the cells to identify the purity of preadipocytes. The over-expression vector of porcine NR1H3 gene was constructed and the NR1H3 siRNA sequences were designed, and then both of them were transfected into porcine preadipocytes. The overexpression and interference efficiency and their effects on expression of key genes related to adipogenic differentiation were investigated by qRT-PCR, Western blot and Oil red O staining methods. The results showed that the expression of NR1H3 in subcutaneous adipose tissues increased as the age increased, the expression level was the highest at 240 days of age and the lowest at 30 days of age, with extremely significant difference between them (P<0.01). Compared with negative control (NC), over-expression (OE) of NR1H3 significantly increased the number of lipid droplets in adipocytes, the expression levels of downstream targets SREBP-1c and ChREBP extremely significantly increased (P<0.01), and the mRNA expression levels of key adipogenic genes FAS, C/EBPβ, PPARγ and FABP4 extremely significantly increased (P<0.01). On the contrary, interfering with NR1H3 significantly reduced the number of lipid droplets. The mRNA expression of downstream target genes and key adipogenic genes extremely significantly reduced (P<0.01), which inhibited the adipogenesis process. The results showed that NR1H3 gene was a positive regulator of the adipogenic differen-tiation of porcine preadipocytes, and affected the differentiation of porcine preadipocytes by regulating the expression of its downstream targets SREBP-1c and ChREBP. These results are of great significance to elucidate the molecular mechanism of fat deposition and improve meat quality.

Key words: pig, NR1H3, porcine preadipocyte, adipogenic differentiation

CLC Number:

S828.2

SHI Mingyue, ZHANG Xuelian, YANG Xiaofen, NIU Jin, XING Jiandong, LU Chang, GAO Pengfei, GUO Xiaohong, LI Bugao, CAO Guoqing. Study on NR1H3 Gene Regulating Differentiation of Porcine Preadipocyte[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2094-2103.

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Study on NR1H3 Gene Regulating Differentiation of Porcine Preadipocyte

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