ADCY2基因表达对延边黄牛脂肪前体细胞成脂分化的影响

doi:10.16431/j.cnki.1671-7236.2020.11.021

Abstract

Abstract: The aim of the experiment was to study the effect of 3',5'-cyclic adenosine monophosphote (cAMP) adenylyl cyclase 2 (ADCY2) gene on adipogenic differentiation of preadipocytes in Yanbian Yellow cattle.The 3-day-old inguinal adipose tissue of Yanbian Yellow cattle was selected for the isolation,culture and induction differentiation of preadipocytes.The interference fragment of ADCY2 gene was designed and the pEX4 overexpression vector was constructed.Adipose cells of normal (Control),interference,and overexpression groups at 0,5,and 10 d were collected,respectively.The mRNA transcriptional levels of peroxisome proliferation activator receptor γ(PPARγ),CCAAT/enhancer binding protein (C/EBP) and ADCY2 in the process of cell differentiation were detected by Real-time quantitative PCR and the protein expression was detected by Western blotting.Oil red O staining was used to detect the change of lipid droplets,and triglyceride content was detected with triglyceride kit.The results showed that during adipogenic differentiation process,compared with the undifferentiated group,the expression of ADCY2 gene increased significantly at the 5th and 10th day (P<0.01),but decreased significantly at the 10th day compared with that of the 5th day (P<0.01).Compared with control group,the mRNA level of ADCY2 gene was increased about 4 000 000 folds in the overexpression group,the number of lipid droplets and the content of triglyceride in adipocytes was extremely increased (P<0.01),the expression of key lipid-forming genes C/EBPα and PPARγ were significantly up-regulated (P<0.01),but the mRNA levels of ADCY2 gene was significantly decreased in the RNA interference group,and the number of lipid droplets and the content of triglyceride in adipocytes decreased significantly(P<0.01),the expression of key lipid-forming genes C/EBPα and PPARγ were significantly down-regulated (P<0.01).Therefore,overexpression of ADCY2 gene could significantly increase the content of lipid droplets and triglyceride in mature adipocytes,and promote the expression of key lipid-forming genes C/EBPα and PPARγ,indicated that ADCY2 gene had a positive regulatory effect on adipocyte differentiation.

Key words: ADCY2 gene; Yanbian Yellow cattle; adipocyte differentiation

CLC Number:

Q952.6

LUO Xiaotong, JIN Xing, YAN Changguo, PIAO Zhengyu. Effect of ADCY2 Gene Expression on Preadipocyte Differentiation in Yanbian Yellow Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(11): 3595-3601.

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Effect of ADCY2 Gene Expression on Preadipocyte Differentiation in Yanbian Yellow Cattle

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