脾酪氨酸激酶参与酿酒酵母甘露聚糖诱导绵羊瘤胃上皮细胞β-防御素-1(SBD-1)的表达

doi:10.11843/j.issn.0366-6964.2019.05.019

Abstract

Abstract:

The aim of this study was to explore whether spleen tyrosine kinase (Syk) is involved in the expression of β-defensin-1 (SBD-1) in ovine ruminal epithelial cells (ORECs) induced by Saccharomyces cerevisiae Mannan (S.c M). Firstly, the expression of Syk in rumen tissues and ORECs was detected by immunohistochemistry, RT-PCR and immunofluorescence. Then, qPCR and Western blot were used to detect the expression of Syk after S.c M stimulation of ORECs, and the phosphorylation level of Syk was detected by Western blot. The ORECs were transfected with three Syk-specific siRNAs (#1, #2 and #3) for 24 h, qPCR was used to detect the expression of Syk mRNA, and the Syk siRNA with the best interference effect was screened. Finally, after treatment of ORECs with the best siRNA and Syk specific inhibitor R406, the expression of SBD-1 was detected by qPCR and ELISA to determine the role of Syk in the expression of SBD-1 induced by S.c M. The results showed that Syk was expressed in ORECs, and the expression level of Syk mRNA and protein were significantly higher in mannan-stimulated ORECs than in unstimulated group (P<0.05). After S.c M stimulation of ORECs for different time (5, 15, 30, 45 and 60 min), Syk was phosphorylated, and the phosphorylation level reached the maximum after 15 min stimulation (P<0.01). In addition, the expression of Syk was decreased after Syk's three specific siRNAs transfected with ORECs, and the inhibitory effect of Syk siRNA#2 was the most significant (P<0.01). Simultaneously, Syk siRNA#2 and R406 significantly reduced S.c M-induced ORECs. Expression of SBD-1 (P<0.01). The above results indicate that Syk is involved in the expression of SBD-1 induced by S.c M.

CLC Number:

S852.4

JIN Xin, ZHANG Man, YANG Yinfeng. Syk is Involved in the Expression of SBD-1 in Ovine Ruminal Epithelial Cells Induced by Saccharomyces cerevisiae Mannan[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(5): 1082-1090.

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Syk is Involved in the Expression of SBD-1 in Ovine Ruminal Epithelial Cells Induced by Saccharomyces cerevisiae Mannan

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