低pH环境对体外原代培养犊牛瘤胃上皮细胞炎性通路的影响

Effects of low environmental p H on expression and activity of the inflammatory cytokines in REC cultured in vitro

本试验旨在研究低p H环境对瘤胃上皮细胞(REC)炎性因子表达的影响。建立犊牛原代REC体外培养模型,在不同p H(5.5和7.2)的培养环境下,采用VFA和NF-κB p65抑制剂PDTC处理。运用Western blot和RT-PCR技术,检测NF-κB p65和IκB蛋白的表达与细胞炎性因子IL-1β、IL-6、TNF-α和NF-κB p65 m RNA表达。结果显示,在低p H环境(p H 5.5)处理组,NF-κB p65和IκB蛋白的表达显著升高;IL-1β、IL-6、TNF-α和NF-κB p65的m RNA表达水平均显著或极显著高于对照组(P<0.05/P<0.01)。组内差异不显著(P>0.05)。

Objective ： to investigate the effects of low environmental pH on expression and activity of the inflammatory cyto- kines in REC cultured in vitro. After 96 h, the cells were treated with different pH media （pH 5.5 or 7.2）for 10 h, and each group with three treatments：0, VFA, and PDTC, respectively. The expression of NF-KB p65 and IKB protein was assessed by Western blot, and the result showed that it increased significantly in pH 5.5 group. The mRNA levels of were detected with RT-PCR, in pHS.5 group, the mRNA levels of TNF-α, IL-6, NF-KB p65 and IL-1β were significantly higher than those of the control group （ P〈0.05/P〈0.01 ）.