Effects of CIDEA Gene on Lipid Metabolism Related Gene Expression and Triglyceride Synthesis in Dairy Cow (Bos taurus) Mammary Epithelial Cells
ZHAO Xin1,2,3,4*, YAO Da-Wei2,3,4*, ZHAO Shu-Ying5, YANG Chun-Lei2,3,4, ZHANG Man2,3,4, CHEN Cheng-Bin1**, MA Yi2,3,4**
1 College of Life Science, Nankai University, Tianjin 300071, China; 2 Institute of Animal Science and Veterinary, Tianjin Academy of Agricultural Sciences, Tianjin 300381, China; 3 Tianjin Key Laboratory of Animal Molecular Breeding and Biotechnology, Tianjin 300381, China; 4 Tianjin Engineering Research Center of Animal Healthy Farming, Tianjin 300381, China; 5 College of Animal Science and Animal Medicine, Tianjin Agricultural University, Tianjin 300384, China
Abstract：CIDEA is a lipid droplet (LD) -associated protein that is highly expressed in the lactating mammary gland, mediates the transformation of small lipid droplets into large lipid droplets and promotes triglyceride (TAG) accumulation. In order to detect the effect of CIDEA gene on lipid metabolism-related gene expression and TAG synthesis in mammary epithelial cells of dairy cows (Bos taurus), the expression of CIDEA gene in mammary tissues of dairy cows during dry milk and lactation was detected by qPCR technique; the synthesis of siRNA targeting CIDEA gene was designed to detect the expression of genes related to lipid metabolism; and the intracellular TAG content was detected by TAG kit. Results showed that the sequence of the CDS region of the full-length 660 bp cow CIDEA gene (GenBank No. MW960011) was obtained by cloning. Bioinformatics analysis revealed that the nucleotide sequences of B. indicus (GenBank No. XM_019986952.1), B. mutus (GenBank No. XM_005892367.1) and Bubalus bubalis (GenBank No. XM_006043025.3) were 99.73%, 99.46% and 98.52%, respectively, and the amino acid sequence homology was more than 95%. The qPCR technique revealed that the expression of CIDEA gene was 1.5 times higher in dairy cows during peak lactation than dry period; siRNA was transfected in dairy cows mammary epithelial cells and the interference efficiency reached over 95%. Interfering with the CIDEA gene significantly downregulated the expression of sterol regulatory element binding protein 1 (SREBF1) and acetyl coenzyme A carboxylase alpha (ACACA), and inhibited the expression of diacylglycerol acyltransferase 1 (DGAT1), DGAT2, lipin 1 (LPIN1), and heart fatty acid binding protein 3 (FABP3), cluster of differentiation 36 (CD36)(P<0.05), and promoted TAG hydrolase (ATGL) expression (P<0.05). After CIDEA interference, a significant decrease in intracellular TAG content was detected (P<0.05). In conclusion, CIDEA plays an important role in regulating genes related to lipid metabolism and TAG synthesis in BMEC. This study further clarifies the regulation mechanism of lipid metabolism in dairy cows, which has important theoretical and practical implications for the healthy development of the dairy industry.
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