组胺对体外培养犊牛原代瘤胃上皮细胞炎性因子的影响  

Effects of Histamine on Inflammatory Cytokines in Calf Ruminal Epithelium Cell Cultured in vitro

作　　者：王婷婷[1] 郭立辉[1] 赵晨旭[1] 王亚洲[1] 刘国文[1] 王哲[1] 

机构地区：[1]吉林大学动物医学学院,长春130062

出　　处：《吉林农业大学学报》2015年第3期363-367,374,共6页Journal of Jilin Agricultural University

基　　金：国家自然科学基金项目(31360630)

摘　　要：为探讨组胺对体外原代培养犊牛瘤胃上皮细胞炎性因子表达及含量的影响。体外培养犊牛瘤胃上皮细胞,在不同p H的培养液(p H 5.5和p H 7.2)中添加不同浓度的组胺(0,0.5,2.5,12.5,62.5μmol/L)。运用实时荧光定量PCR和ELISA方法,检测组胺对瘤胃上皮细胞炎性因子白介素1β(IL-1β)、白介素6(IL-6)和肿瘤坏死因子α(TNF-α)m RNA表达及其含量的影响。结果表明:p H 5.5的培养液组,TNF-α、IL-1β和IL-6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);TNF-α、IL-6和IL-1β的含量在2.5,12.5,62.5μmol/L组胺处理组显著高于对照组(P<0.05,P<0.01)。p H 7.2的培养液组,TNF-α和IL-6 m RNA表达水平在2.5,12.5μmol/L组胺处理组显著高于对照组(P<0.05,P<0.01),IL-1βm RNA表达水平在2.5,12.5,62.5μmol/L组显著高于对照组(P<0.05,P<0.01);TNF-α含量在62.5μmol/L组极显著高于对照组(P<0.01),IL-6在2.5,12.5,62.5μmol/L组显著高于对照组(P<0.05,P<0.01),IL-1β在12.5,62.5μmol/L组极显著高于对照组(P<0.01)。在组胺各浓度组,TNF-α、IL-6和IL-1βm RNA表达水平在p H5.5条件下均显著高于p H 7.2(P<0.05,P<0.01),但含量差异不显著。低p H条件下,高浓度组胺能增加犊牛瘤胃上皮细胞炎性反应。This study aimed to investigate the effects of histamine on expression and content of in- flammatory cytokines in calf ruminal epithelium cells cultured in vitro. After 96 h culture, ruminal epithelium cells were treated with different concentrations of histamine （0, 0. 5, 2.5, 12.5 and 62. 5 μmol/L） and pH. The mRNA levels were detected with real-time fluorescent quantitative PCR. The content of TNF-a, IL-6 and IL-1β were detected with enzyme-linked immunosorbent assay. In the pH 5.5 medium, the mRNA levels of TNF-a, IL-6 and IL- 1β were significantly higher than in the control group, and TNF-a, IL-6 and IL-1β content was significantly increased in 2. 5 μmol/L,12. 5 μmol/L and 62. 5 p^mol/L histamine treatment groups （ P〈0. 05, P〈0. 01 ）. In the pH 7.2 medium, TNF-a and IL-6 mRNA levels were significantly higher in the 2.5 μmol/L, 12.5 μmol/L and 62.5 μmol/L histamine treatment groups （P〈0. 05 ,P〈0. 01）. TNF-ot content was significantly increased in 62. 5 μmol/L group （P〈0. 05 ,P〈0.01 ）. IL-6 content was significantly increased from 2. 5 μmol/L to 62.5 μmol/L groups （P〈0. 05 ,P〈0. 01 ）. IL-1β content was markedly increased in the 12. 5 μmol/L and 62.5 μmol/L groups （P〈0.05, P〈0. 01）. In histamine treatment groups, TNF-a, IL-6 and IL-1β mRNA levels were significantly higher in the pH 5.5 treatment medium than in the pH 7.2 medium （P〈0. 05,P〈0. 01）. However, there was no significant change in the content of TNF-a, IL-6 and IL-1β. These results indicate that high levels of histamine increase in low pH conditions inflammatory response in ruminal epithelium cells.

关 键 词：犊牛 组胺 瘤胃上皮细胞 炎性因子 pH 

分 类 号：S856.5[农业科学—临床兽医学]