干扰HSD17B4基因对水牛乳腺上皮细胞乳脂和酪蛋白的影响

doi:10.16431/j.cnki.1671-7236.2021.09.003

Abstract

Abstract: The expression changes of casein content, triglyceride content and fatty acid-related genes were detected by transfecting small interfering-hydroxysteroid 17-beta dehydrogenase Ⅳ(si-HSD17B4) in buffalo mammary epithelial cell, which were revealing the effects of interfering with the HSD17B4 gene on milk fats and caseins in buffalo mammary epithelial cell. The relative expressions of HSD17B4 mRNA in various tissues of buffalo were detected by Real-time quantitative PCR. 3 HSD17B4 siRNA fragments of buffalo and one control si-HSD17B4-nc (si-nc) fragment were synthesized and then the optimal fragments and time of the interference were selected. Caseins were detected by casein kits after si-HSD17B4 transfected buffalo mammary epithelial cell. Then triglycerides were detected by triglyceride kit and oil red O staining, and the effects of HSD17B4 gene interference on the gene expression of the triglycerides synthesis and fatty acid synthesis and oxidation. The results showed that HSD17B4 gene was relatively highly expressed in buffalo heart and ovary, and was significantly higher than other tissue(P<0.05). 3 HSD17B4 siRNA fragments could effectively decrease the expression of the HSD17B4 gene, and the si-HSD17B4-1 was the best interference fragment, the relative expression of HSD17B4 mRNA was extremely significantly lower than that in the control group (P<0.01). The optimum interference time was 24 h. Casein test showed no effect of interfering HSD17B4 on casein synthesis in buffalo mammary epithelial cell. Triglyceride detections showed an increase in triglyceride content in cells after interfering with the HSD17B4 gene with significant differences (P<0.05). The oil red O tests showed a significant increase in fat droplets in cells after interfering with HSD17B4 gene. The results of Real-time quantitative PCR showed that interference with the HSD17B4 gene, the relative expressions of FABP3, ACSL1, DGAT1, DGAT2, PLIN2 and BTN1A1 gene were increased by 9.28 (P<0.01), 1.36 (P<0.05), 1.17 (P<0.05), 1.83 (P<0.01), 1.60 (P<0.01) and 2.17 (P<0.01) times, respectively. The relative expressions of PPARA, SREBP1C, SCD, ACSS2, ACACA, AGPAT6, LPIN1, XDH, ABCD1, ACOX2, EHHADH and SCP2 genes decreased to 50.55% (P>0.05), 61.15% (P<0.01), 84.91% (P<0.01), 89.34% (P<0.01), 21.88% (P<0.01), 86.48% (P<0.01), 25.35% (P<0.01), 27.24% (P<0.01), 22.17% (P<0.01), 62.70% (P<0.01) and 70.33% (P<0.01), respectively. The results showed that the HSD17B4 gene was widely expressed in buffalo tissue. si-HSD17B4 effectively reduced the expression of the HSD17B4 gene in buffalo mammary epithelial cell, and it had no effect of interfering with the HSD17B4 gene on the caseins in mammary epithelial cell, but it would increase the expressions of FABP3, ACSL1, DGAT1, DGAT2 genes to promote the synthesis of triglycerides while decrease the expression of ABCD1, ACOX2, EHADH, SCP2 genes to reduce the β-oxidation of fatty acids.

Key words: buffalo; mammary epithelial cell; HSD17B4; fatty acid; casein

CLC Number:

S823.9

DUAN Anqin, LU Xingrong, MA Xiaoya, LIANG Shasha, DENG Tingxian. Effect of HSD17B4 Gene Interference on Milk Fat and Casein in Buffalo Mammary Epithelial Cell[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(9): 3128-3137.

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Effect of HSD17B4 Gene Interference on Milk Fat and Casein in Buffalo Mammary Epithelial Cell

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