干扰CREB基因对奶山羊乳腺上皮细胞脂质合成相关基因表达及甘油三酯合成的影响

doi:10.11843/j.issn.0366-6964.2019.12.005

Abstract

Abstract: The study aimed to screen out the siRNA of cAMP response element binding protein (CREB) gene in dairy goats, and reveal the effect of interfering with CREB on the expression of milk fat synthesis related genes and triglyceride synthesis in mammary epithelial cells. The complete CDS region of CREB gene was amplified from the mammary gland tissue of Xinong Saanen dairy goat by qRT-PCR. Sequence and CREB expression analysis in different lactation periods were performed. The siRNA targeting the CREB gene was synthesized. The effective siRNA was screened by fluorescent quantitative PCR, at the same time, the expression of genes related to lipid synthesis was detected. The intracellular triglyceride content was detected using a kit. The results showed that:1) The CDS region of the CREB gene with 984 bp in dairy goat was cloned (GenBank accession number:MK158073) and analyzed by bioinformatics. 2) The expression level of CREB gene in the mammary gland of goats in lactating period was 1.93 times higher than that of goat in the dry period(P<0.05). 3) The effective siRNA targeting CREB gene was successfully screened, the interference efficiency was 72% (P<0.01),meanwhile transfecting it into the mammary gland epithelial cells of dairy goats. The expression of lipid synthesis-related genes were detected by qRT-PCR. Compared with the control group, the expression of FASN, ACACA, SCD1, FABP3,LPL, CPT1B,GPAM and DGAT2 genes was significantly inhibited after interfering with CREB gene (P<0.05), and the expression level of HSL was significantly increased(P<0.05),as well as the content of intracellular triglyceride was significantly inhibited(P<0.05). In summary, CREB gene maybe play an important role in goat milk fat synthesis by regulating the expression of lipid metabolism related genes and the content of triglyceride in primary mammary epithelial cells of dairy goats.

Key words: milk goat, mammary epithelial cell, CREB, RNA interference, lipid synthesis, triglyceride

CLC Number:

S287
S813.3

MA Jing, YAO Dawei, YANG Chunlei, LI Qiuling, WANG Tianzhen, CHEN Chengbin, SONG Wenqin, MA Yi. Effects of Interfering with CREB Gene on Lipid Synthesis Related Gene Expression and Triacylglycerol Synthesis in Mammary Epithelial Cells of Dairy Goats[J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50(12): 2413-2421.

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Effects of Interfering with CREB Gene on Lipid Synthesis Related Gene Expression and Triacylglycerol Synthesis in Mammary Epithelial Cells of Dairy Goats

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