大黄上清液联合母乳对Caco-2炎症细胞核转录因子表达的影响

The Impact of Supernatant Fluid of Rhubarb Combined with Human Breast Milk on the Expression of Nuclear Transcription Factor in Caco-2 Inflammatory Cell

目的探讨大黄上清液及母乳对坏死性小肠结肠炎(NEC)炎症细胞的分子保护机制。方法通过运用炎症因子TNF-α诱导NEC肠道炎症细胞模型,分别加入常规培养液(A组)、1%的大黄上清液(B组)、1%的乳清(C组)、1%大黄上清液加1%乳清(D组),采用免疫细胞化学测定各组NF-κB P65蛋白表达量及RTPCR检测法测定各组NF-κB P65蛋白下游产物IL-1βmRNA的相对表达量。结果与A组比较,B、C、D组均可下调NF-κB P65蛋白表达,下调程度D组最大,C组次之,B组最小。与A组比较,B、C、D组均可抑制IL-1βmRNA表达(均P<0.01);D组较B组效果明显(P<0.01);D组较C组作用强,差异有显著统计学意义(P<0.05),C较B组作用强,但差异无统计学意义(P>0.05)。结论大黄上清液及乳清可抑制Caco-2炎症细胞核转录因子NF-κB P65蛋白及其下游产物IL-1βmRNA的表达,此机制可能为大黄及母乳防治NEC的作用机制之一。

Objective：To study the effectiveness and the mechanisms of rhubarb and breast milk preventing Necrotizing enterocolitis （NEC） through the level of cellular and molecular biology to provide the evidence for clinical application.Methods：We induced Caco-2 inflammation cell model of NEC with inflammatory factor TNF-α.The model group was divided into four groups：A-conventional culture added,B-1％ rhubarb supernatant fluid,C-l％ breast milk supernatant fluid,D-the combination of 1％ rhubarb supernatant fluid and 1％ breast milk supernatant fluid.We detected the expression of NF-κB P65 with immunocytochemical method and the relative expression of IL-1βmRNA which is downstream product of NF-κB P65 with RT-PCR method.Results：We discovered B,C,D group could obviously inhibit the expression of NF-κB P65,compared with A group.The inhibition effect of D group was best,C group second and B group last.We discovered B,C,D group with P＜ 0.01 could obviously inhibit the expression of IL-1β mRNA,compared with A group.Compared with B group,D group with P＜ 0.01 had a better effect;compared with C group,D group had better effect with significant statistical difference （P＜ 0.05）;compared with B group,C group had better effect,but the difference had no statistical significance （P＞ 0.05）.Conclusion：Rhubarb supernatant fluid and breast milk supernatant fluid can inhibit the expression of NF-κB P65 and its downstream product IL-1β mRNA of inflammatory Caco-2 cells,which may be one of the mechanisms of rhubarb and breast milk preventing NEC.