LPIN1基因对水牛乳腺上皮细胞甘油三酯和脂肪酸合成的影响

doi:10.16431/j.cnki.1671-7236.2020.10.012

Abstract

Abstract: The purpose of this study was to construct the adenovirus pDC316-LPIN1-eGFP vector and package the Ad-LPIN1 adenovirus particles,in order to explore the effect of LPIN1 gene on triglyceride and fatty acid synthesis in buffalo mammary epithelial cells.The expression of LPIN1 gene and lipin1 protein in buffalo mammary epithelial cell were detected by Real-time quantitative PCR and immunofluorescence (IF) after Ad-LPIN1 infected.The triglyceride was detected by oil red O staining and triglyceride measurement and the genes expression of triglyceride synthesis and fatty acid synthesis after overexpressing LPIN1 were detected by Real-time quantitative PCR.The results showed that high-quality Ad-LPIN1 adenovirus particles were packaged with the pDC316-LPIN1-eGFP vector.Ad-LPIN1 adenovirus particles infected the buffalo mammary epithelial cell (MOI=50),LPIN1 mRNA was relatively up-regulated more than 5 000-fold,which was extremely significant difference (P<0.01).lipin1 protein expression was up-regulated and mainly expressed in the nucleus.Oil red O staining results showed that lipid droplets in cells were significantly reduced after overexpressing LPIN1 gene.The results of triglyceride measurement showed that the triglyceride content in cells decreased after overexpressing LPIN1 gene,and the difference was extremely significant (P<0.01).The mRNA of peroxisome proliferator activated receptor gama (PPARγ),peroxisome proliferator activated receptor alpha (PPARα) and sterol regulatory element binding transcription factor 1 (SREBP1) after overexpressing LPIN1 gene were significantly or extremely significant increased (P<0.05;P<0.01).And the mRNA of stearoyl-CoA desaturase (SCD),fatty acid synthase (FASN),acyl-CoA synthetase long chain family member 1 (ACSL1) and acetyl-CoA carboxylase alpha (ACACA) were decreased to 83.17%,51.30%,44.24%,41.18% (P<0.01),respectively.The above-mentioned results showed that adenovirus efficiently mediated the expression of LPIN1 gene in buffalo mammary epithelial cell,and increased the expression of lipin1 protein in the nucleus.lipin1 protein,as a common transcription activation factor,inhibited the synthesis of triglycerides by down-regulating mRNA of ACACA,FASN,ACSL1 and SCD genes.

Key words: buffalo; mammary epithelial cell; LPIN1 gene; triglyceride; fatty acid

CLC Number:

S823.8⁺3

DUAN Anqin, LU Xingrong, MA Xiaoya, LIANG Shasha, DENG Tingxian. Effect of LPIN1 Gene on Triglyceride and Fatty Acid Synthesis in Buffalo Mammary Epithelial Cell[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(10): 3140-3148.

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Effect of LPIN1 Gene on Triglyceride and Fatty Acid Synthesis in Buffalo Mammary Epithelial Cell

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