Regulation of bta-miRNA29d-3p on Lipid Accumulation via GPAM in Bovine Mammary Epithelial Cells

MicroRNAs (miRNAs) are small RNA molecules consisting of approximately 22 nucleotides that are engaged in the regulation of various bio-processes. There is growing evidence that miR-29 is a key regulator of hepatic lipid metabolism. Mimics and inhibitors of bta-miRNA29d-3p were transiently transfected in bovine mammary epithelial cells (BMECs) to reveal the regulation of bta-miRNA29d-3p on lipid accumulation in BMECs. Results showed that overexpression of bta-miRNA29d-3p significantly inhibited the expression of genes related to triglyceride (TAG) synthesis, namely DGAT1 and mitochondrial glycerol-3-phosphate acyltransferase (GPAM, p < 0.01) and down-regulated TAG levels in cells (p < 0.05). The expression of fatty acid synthesis and desaturation-related genes FASN, SCD1, and ACACA, and transcription factor SREBF1 also decreased. Interference of bta-miRNA29d-3p significantly increased the expression of GPAM, DGAT1, FASN, SCD1, ACACA, and SREBF1 (p < 0.01), and significantly upregulated the concentration of TAG in cells. Furthermore, a luciferase reporter assay confirmed that GPAM is a direct target of bta-miRNA29d-3p. In summary, bta-miRNA29d-3p modulates fatty acid metabolism and TAG synthesis by regulating genes related to lipid metabolism in BMECs and targeting GPAM. Thus, bta-miRNA29d-3p plays an important role in controlling mammary lipid synthesis in cows.

[1]  Zhihui Zhao,et al.  Transcriptome analysis of CRISPR/Cas9-mediated GPAM-/- in bovine mammary epithelial cell-line unravelled the effects of GPAM gene on lipid metabolism. , 2022, Gene.

[2]  Zhangping Yang,et al.  circ01592 regulates unsaturated fatty acid metabolism through adsorbing miR-218 in bovine mammary epithelial cells. , 2021, Food & function.

[3]  C. Farquharson,et al.  The role of miR‐29 family in disease , 2021, Journal of cellular biochemistry.

[4]  Runjun Yang,et al.  Effects of polymorphism of the GPAM gene on milk quality traits and its relation to triglyceride metabolism in bovine mammary epithelial cells of dairy cattle , 2021, Archives animal breeding.

[5]  V. Boussiotis,et al.  The role of Peroxisome Proliferator-Activated Receptors (PPAR) in immune responses. , 2020, Metabolism: clinical and experimental.

[6]  Fangting Zhou,et al.  Isolation, identification, expression and subcellular localization of PPARG gene in buffalo mammary gland. , 2020, Gene.

[7]  X. An,et al.  MiR-29 regulates the function of goat granulosa cell by targeting PTX3 via the PI3K/AKT/mTOR and Erk1/2 signaling pathways , 2020, The Journal of Steroid Biochemistry and Molecular Biology.

[8]  L. Hasoňová,et al.  Role of Fatty Acids in Milk Fat and the Influence of Selected Factors on Their Variability—A Review , 2018, Molecules.

[9]  A. Brunner,et al.  Cardiac myocyte miR-29 promotes pathological remodeling of the heart by activating Wnt signaling , 2017, Nature Communications.

[10]  Junya Li,et al.  Bovine lipid metabolism related gene GPAM: Molecular characterization, function identification, and association analysis with fat deposition traits. , 2017, Gene.

[11]  D. O'Gorman,et al.  Altered miR-29 Expression in Type 2 Diabetes Influences Glucose and Lipid Metabolism in Skeletal Muscle , 2017, Diabetes.

[12]  A. Chakravarti,et al.  Feedback Loop Regulation of SCAP/SREBP-1 by miR-29 Modulates EGFR Signaling-Driven Glioblastoma Growth. , 2016, Cell reports.

[13]  Ming Li,et al.  Bioinformatics analysis of hepatitis C virus genotype 2a-induced human hepatocellular carcinoma in Huh7 cells , 2016, OncoTargets and therapy.

[14]  Lixin Liu,et al.  Epigenetic Regulation of miR‐29s Affects the Lactation Activity of Dairy Cow Mammary Epithelial Cells , 2015, Journal of cellular physiology.

[15]  P. Sethupathy,et al.  Inhibition of miR-29 has a significant lipid-lowering benefit through suppression of lipogenic programs in liver , 2015, Scientific Reports.

[16]  P. Sethupathy,et al.  MicroRNA-29 Fine-tunes the Expression of Key FOXA2-Activated Lipid Metabolism Genes and Is Dysregulated in Animal Models of Insulin Resistance and Diabetes , 2014, Diabetes.

[17]  D. Bauman,et al.  Liver x receptors stimulate lipogenesis in bovine mammary epithelial cell culture but do not appear to be involved in diet‐induced milk fat depression in cows , 2014, Physiological reports.

[18]  S. Bensinger,et al.  Liver X receptor and peroxisome proliferator‐activated receptor as integrators of lipid homeostasis and immunity , 2012, Immunological reviews.

[19]  Timothy F. Osborne,et al.  SREBPs: metabolic integrators in physiology and metabolism , 2012, Trends in Endocrinology & Metabolism.

[20]  R. E. Everts,et al.  Gene network and pathway analysis of bovine mammary tissue challenged with Streptococcus uberis reveals induction of cell proliferation and inhibition of PPARγ signaling as potential mechanism for the negative relationships between immune response and lipid metabolism , 2009, BMC Genomics.

[21]  J. Loor,et al.  Peroxisome proliferator-activated receptor-gamma activation and long-chain fatty acids alter lipogenic gene networks in bovine mammary epithelial cells to various extents. , 2009, Journal of dairy science.

[22]  J. Loor,et al.  Adipose tissue lipogenic gene networks due to lipid feeding and milk fat depression in lactating cows. , 2009, Journal of dairy science.

[23]  J. Loor,et al.  Gene networks driving bovine milk fat synthesis during the lactation cycle , 2008, BMC Genomics.

[24]  Aibin He,et al.  Overexpression of micro ribonucleic acid 29, highly up-regulated in diabetic rats, leads to insulin resistance in 3T3-L1 adipocytes. , 2007, Molecular endocrinology.

[25]  J. Loor,et al.  Identification of reference genes for quantitative real-time PCR in the bovine mammary gland during the lactation cycle. , 2007, Physiological genomics.

[26]  S. O’Rahilly,et al.  PPARγ and human metabolic disease , 2006 .

[27]  R. E. Everts,et al.  Temporal gene expression profiling of liver from periparturient dairy cows reveals complex adaptive mechanisms in hepatic function. , 2005, Physiological genomics.

[28]  Lothar Hennighausen,et al.  Information networks in the mammary gland , 2005, Nature Reviews Molecular Cell Biology.

[29]  Robert V Farese,et al.  DGAT1 Is Not Essential for Intestinal Triacylglycerol Absorption or Chylomicron Synthesis* , 2002, The Journal of Biological Chemistry.

[30]  E. Saggerson,et al.  Glycerolipid metabolizing enzymes in rat ventricle and in cardiac myocytes. , 1997, Biochimica et biophysica acta.

[31]  Z. Sh.  Research Progress on Regulation Effect of microRNA on Mammary Gland Development and Lactation , 2015 .

[32]  R. Leotta,et al.  MILK FATTY ACIDS FROM DIFFERENT HORSE BREEDS COMPARED WITH COW, GOAT AND HUMAN MILK , 2012, Macedonian Journal of Animal Science.