Laboratory of Animal Physiology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
Research Faculty of Agriculture, Hokkaido University, Sapporo, Hokkaido, Japan.
Anim Sci J. 2020 Jan-Dec;91(1):e13422. doi: 10.1111/asj.13422.
The aim of this study was to identify factors that regulate ruminal epithelial insulin-like growth factor-binding protein (IGFBP) expression and determine its role in rumen epithelial cell proliferation. Primary bovine rumen epithelial cells (BREC) were incubated with short-chain fatty acids (SCFAs) at pH 7.4 or 5.6, lactate, lipopolysaccharide (LPS), insulin-like growth factor-I (IGF-I), -II (IGF-II), or recombinant bovine IGFBP2 (rbIGFBP2). The mRNA expression levels of IGFBP in BREC were analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation rate of BREC was analyzed using a WST-1 assay. IGFBP2 gene expression tended to be lower with SCFA treatment (p < .1), and IGFBP6 gene expression was significantly lower with SCFA treatment (p < .05). IGFBP3 and IGFBP6 gene expression tended to be higher with d-Lactate treatment (p < .1). IGFBP3 gene expression was significantly higher (p < .05) with LPS treatment. BREC treated with IGF-I grew more rapidly than vehicle control-treated cells (p < .01); however, recombinant bovine rbIGFBP2 inhibited IGF-I-induced proliferation. IGF-II and/or rbIGFBP2 did not affect BREC proliferation. Taken together, SCFA treatment decreased IGFBP2 and IGFBP6 expression in rumen epithelial cells, and lower expression of these IGFBP might promote rumen epithelial cell proliferation by facilitating IGF-I.
本研究旨在确定调节瘤胃上皮胰岛素样生长因子结合蛋白 (IGFBP) 表达的因素,并确定其在瘤胃上皮细胞增殖中的作用。将原代牛瘤胃上皮细胞 (BREC) 在 pH 7.4 或 5.6、乳酸盐、脂多糖 (LPS)、胰岛素样生长因子-I (IGF-I)、-II (IGF-II) 或重组牛 IGFBP2 (rbIGFBP2) 下孵育。使用实时定量聚合酶链反应 (qRT-PCR) 分析 BREC 中 IGFBP 的 mRNA 表达水平。使用 WST-1 测定法分析 BREC 的增殖率。SCFA 处理后 IGFBP2 基因表达趋于降低(p<.1),SCFA 处理后 IGFBP6 基因表达显著降低(p<.05)。d-乳酸盐处理后 IGFBP3 和 IGFBP6 基因表达趋于升高(p<.1)。LPS 处理后 IGFBP3 基因表达显著升高(p<.05)。与载体对照处理的细胞相比,用 IGF-I 处理的 BREC 生长更快(p<.01);然而,重组牛 rbIGFBP2 抑制 IGF-I 诱导的增殖。IGF-II 和/或 rbIGFBP2 不影响 BREC 的增殖。总之,SCFA 处理降低了瘤胃上皮细胞中 IGFBP2 和 IGFBP6 的表达,这些 IGFBP 的低表达可能通过促进 IGF-I 来促进瘤胃上皮细胞增殖。