猪FUT2基因沉默对其所在通路基因表达及小肠上皮细胞E.coli F18黏附能力的影响

doi:10.11843/j.issn.0366-6964.2017.11.004

Abstract

Abstract:

The purpose of this study was to further explore the function of FUT2 and its regulation mechanisms in swine epithelial cells infected by E. coli F18 at the cellular level, which provide theoretical basis for molecular breeding of improving the resistance of piglets to E. coli F18. The expression differences of FUT2 gene in intestinal epithelial cells before and after E. coli F18ab, F18ac infection and lipopolysaccharide(LPS) induction were detected by Real-time PCR. Then, the Western blotting was performed to identify the protein expression differences of FUT2 in intestinal epithelial cells before and after E. coli F18ab and F18ac infection. Meanwhile the lentiviral interference vectors of porcine FUT2 were designed and constructed, and the intestinal epithelial cells line with FUT2 gene silencing was obtained. The effects of FUT2 gene silencing on the mRNA expression of key genes (FUT1, ST3GAL1, HEXA, HEXB, B3GALNT1, NAGA) involved in glycosphingolipid biosynthesis-globo series pathway and on E. coli F18 adhesion to intestinal epithelial cells were further analyzed. The results showed that the FUT2 expression levels in intestinal epithelial cells were significantly increased after E. coli infection and LPS induction (P<0.01), and the FUT2 expression level in intestinal epithelial cells was also increased after E. coli infection. Moreover, the lentiviral interference vectors of porcine FUT2 were successfully constructed, and the intestinal epithelial cells line with FUT2 gene silencing were obtained. After FUT2 gene silencing, the expression level of key genes in glycosphingolipid biosynthesis-globo series pathway were decreased at different degrees. The expression level of FUT1 after FUT2 silencing was significantly reduced (P<0.05), and the expression levels of ST3GAL1, HEXA, HEXB were extremely significantly reduced (P<0.01), while the expression levels of B3GALNT1 and NAGA genes showed no significant changes. In addition, the adhesion ability of E. coli F18 to swine epithelial cells was decreased significantly (P<0.05). These results show that the lower expression level of FUT2 may hinder the formation of E.coli F18 receptor, and thus elevate the resistance capability of porcine small intestine epithelial cells to E.coli F18 infection. Our findings can also provide theoretical foundations and evidence for further investigating on the mechanisms of glycosphingolipid biosynthesis-globo series pathway in resistance of pigs to E. coli infection.

CLC Number:

S828.2

SUN Li, ZONG Qiu-fang, WU Sen, WU Jia-yun, WU Sheng-long, BAO Wen-bin. The Effects of FUT2 Gene Silencing on the Expression of Genes Involved in Glycosphingolipid Biosynthesis-globo Series Pathway and the Adhesion of E. coli F18 to Small Intestinal Epithelial Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(11): 2034-2045.

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The Effects of FUT2 Gene Silencing on the Expression of Genes Involved in Glycosphingolipid Biosynthesis-globo Series Pathway and the Adhesion of E. coli F18 to Small Intestinal Epithelial Cells

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