小尾寒羊前体脂肪细胞分化过程相关基因表达规律的研究

doi:10.16431/j.cnki.1671-7236.2019.07.018

Abstract

Abstract:

In order to explore the changes of related genes during adipocyte differentiation of Small-tail Han sheep,white inguinal adipose tissuees of two-month-old Small-tail Han sheep were collected and preadipocytes were isolated by enzymatic digestion in vitro.After the cultured preadipocytes were covered with cell plates,the cells were induced to differentiate into mature adipocytes by induction solution Ⅰ and induction solution Ⅱ,respectively.Oil red O staining was used to verify adipocytes and detect lipid droplets.Total RNA was extracted from 70% and 90% of the proliferative cells,48 h of induction Ⅰ,48 h of induction Ⅱ and 48 h of complete culture medium (2,4,6,8,10 d,respectively),and was retranscribed into DNA.Real-time quantitative PCR (RT-qPCR) was used to detect genes PPARγ,C/EBPα,LPL,SREBP1,KLF5,KLF6,FABP4,STAT5,ACSS2,IGF1,ADD1,FOXO1,ACACA,DGAT1 and CPT1A that had been shown to be involved in the differentiation of human and mouse preadipocytes to explore their expression in the differentiation of Small-tail Han sheep.The results showed that the preadipocytes were successfully separated and induced to become mature adipocytes with obvious lipid droplets inside the cells;RT-qPCR assay showed that the expressions of above genes had significant fluctuations in the cell differentiation stage,and the peak time was different;The peak expression of C/EBPα and FOXO1 appeared on the 6th day,which may play an important role in the early stage of cell differentiation.PPARγ,LPL,SREBP1,KLF5,KLF6,FABP4,STAT5,ADD1 and ACSS2 gene peaks appeared on the 8th day,but the expression multiples and trends were different;The expression of ACACA gene fluctuated up and down.The peaks of IGF1 and DGAT1 appeared on the 10th day.The expression of CPT1A was declining;The expression multiple of FABP4 gene was significantly higher than that of other genes.This study completely detected the expression regulation of key genes in the differentiation process of Small-tail Han sheep preadipocytes.The results could provide theoretical reference for exploring the molecular mechanism of Small-tail Han sheep adipose differentiation,discovering new key genes involved in adipose differentiation and improving the intermuscular fat content of Small-tail Han sheep.

Key words: Small-tail Han sheep; preadipocytes; isolation and culture; differentiation; adipocytes; gene expression

CLC Number:

S811.3

XIAO Cheng, JIN Haiguo, WEI Tian, CAO Yang. Study on the Expression of Genes Related to the Differentiation Process of Preadipocytes in Small-tail Han Sheep[J]. , 2019, 46(7): 2030-2037.

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Study on the Expression of Genes Related to the Differentiation Process of Preadipocytes in Small-tail Han Sheep

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