黑酵母β-1,3/1,6-葡聚糖对感染肠炎沙门氏菌Caco-2细胞促炎症和抗炎症细胞因子基因mRNA表达水平的影响

Sophy Beta-1,3 /1,6-Glucan Affects mRNA Expression Levels of Pro-Inflammatory and Anti-Inflammatory Cytokine Genes in Caco-2 Cells Challenged with Salmonella enteritidis

本试验采用实时荧光定量PCR分析黑酵母β-1,3/1,6-葡聚糖对肠炎沙门氏菌(Salmonella enteritidis)诱导Caco-2细胞表达细胞因子基因mRNA表达水平的影响。试验选用Caco-2细胞,分成4个组:对照组(A组)、肠炎沙门氏菌感染组(B组)、β-1,3/1,6-葡聚糖组(C组)和β-1,3/1,6-葡聚糖+肠炎沙门氏菌共培养组(D组)。C、D组用黑酵母β-1,3/1,6-葡聚糖(50μg/mL)预处理24 h,然后B、D组各孔接种1×108CFU/mL肠炎沙门氏菌处理3 h。实时荧光定量PCR方法分析细胞肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-8(IL-8)、白细胞介素-10(IL-10)和转化生长因子-β(TGF-β)基因mRNA表达水平。结果表明:肠炎沙门氏菌和β-1,3/1,6-葡聚糖均极显著地上调了IL-1β、TNF-α、IL-8和IL-10基因mRNA表达水平(P<0.01),但对TG F-β基因mRNA表达水平无显著影响(P>0.05)。与肠炎沙门氏菌感染组相比,添加β-1,3/1,6-葡聚糖可显著地下调肠炎沙门氏菌诱导Caco-2细胞的细胞因子IL-1β、IL-8和TNF-α基因mRNA表达水平(P<0.05),并且显著上调IL-10、TGF-β基因mRNA表达水平(P<0.05)。由此可知,黑酵母β-1,3/1,6-葡聚糖能抑制肠炎沙门氏菌感染引起的肠道上皮细胞免疫炎症反应。

The aim of this study was to investigate the effects of Sophy β-1,3/1,6-glucan on the mRNA expression levels of cytokine genes in Caco-2 cells challenged with Salmonella enteritidis by using real-time fluorescence quantitative PCR method.The Caco-2 cells were treated by minimum essential medium （MEM） without antibiotics （group A）,Salmonella enteritidis （group B）,β-1,3/1,6-glucan （50 μg/mL,group C）,Salmonella enteritidis and β-1,3/1,6-glucan （50 μg/mL,group D）,respectively.The Caco-2 cells in groups C and D were pre-incubated with β-1,3/1,6-glucan （50 μg/mL） for 24 hours,and then Caco-2 cells in groups B and D were treated with Salmonella enteritidis （1 × 108 CFU/mL） for 3 hours.The mRNA expression levels of tumor necrosis factor-α （TNF-α）,interleukin-1β （IL-1β）,interleukin-8 （1L-8）,interleukin-10 （IL-10） and transforming growth factor-β （TGF-β） genes were detected with real-time fluorescence quantitative PCR.The results showed that the mRNA expression levels of IL-1β,TNF-α,IL-8 and IL-10 genes were significantly up-regulated by Salmonella enteritidis challenge or Sophy β-1,3/1,6-glucan treatment alone （P 〈 0.01）,but the mRNA expression level of TGF-β gene was not affected （P 〉 0.05）.Compared with group B,the supplementation of Sophy β-1,3/1,6-glucan （group D） significantly down-regulated the mRNA expression levels of IL-1β,IL-8 and TNF-α genes in Caco-2 cells challenged by Salmonella enteritidis （P 〈0.05）,and significantly up-regulated mRNA expression levels of IL-10 and TGF-β genes （P 〈 0.05）.This study indicates that β-1,3/1,6-glucan can suppress the inflammatory response of intestinal epithelial cells challenged by Salmonella enteritidis.