母乳对Caco-2细胞缺氧复氧损伤模型NF-κB p65表达的影响  

Impact of Breast Milk on the Expression of NF-κB p65 in Caco-2 Cells Model with Hypoxia/Reoxygenation Injury

作　　者：陆玲[1] 王宇军[2] 冯伟伟[2] 毕明远[2] 阮为勇[2] 

机构地区：[1]南京中医药大学,210023 [2]江苏省中西医结合医院儿科,南京210028

出　　处：《医学研究杂志》2015年第7期135-138,共4页Journal of Medical Research

基　　金：江苏省中西医结合医院基金资助项目(RC1102)

摘　　要：目的通过研究母乳对Caco-2细胞缺氧复氧损伤模型NF-κB p65表达的影响,以期探讨母乳对坏死性小肠结肠炎(necrotizing enterocolitis,NEC)的作用机制。方法实验分为对照组和缺氧复氧损伤模型组,两组各有常规培养液组(MEM)、未加热乳清组(5%BM)和加热乳清组(5%BMb)。造模后向两组分别加入常规培养液、未加热乳清、加热乳清,继续培养6h后分别用RT-PCR检测法和免疫荧光法检测各组细胞中NF-κB p65表达情况。结果 RT-PCR检测结果显示对照组和模型组中,5%乳清和5%加热乳清均能抑制NF-κB p65 m RNA的表达(P<0.01),模型组中5%乳清抑制NF-κB p65 m RNA表达的作用较5%加热乳清更明显(P<0.05);免疫荧光结果显示,缺氧复氧损伤后Caco-2细胞质内的NF-κB p65被激活进入细胞核表达,5%乳清可以抑制NF-κB p65入核表达,并且较加热乳清作用更明显。结论新鲜母乳可通过抑制NF-κB p65的表达防治小肠上皮细胞缺血再灌注损伤,其可能是母乳防治新生儿坏死性小肠结肠炎机制之一。Objective To study the mechanisms of breast milk in preventing necrotizing enterocolitis （ NEC） through determining the impact of breast milk on the expression of NF-κB p65 in Caco-2 cells model with hypoxia/reoxygenation injury.Methods The study had control groups and model groups.They were divided into three groups（ MEM,5%BM and 5%BMb） .We added conventional cul-ture fluid,5%breast milk supernatant fluid and 5%breast milk supernatant fluid boiled into the three groups respectively after the model was builded.All cells continually cultured six hours, then the expression of NF-κB p65 in Caco-2 cells were detected with RT-PCR and immunofluorescence method.Results RT-PCR method showed that in control groups and model groups 5%breast milk supernatant fluid boiled or unboiled could inhibit the expression of NF-κB p65 mRNA（P〈0.01）.Compared with 5%breast milk supernatant fluid boiled,the inhibitory effect of 5% breast milk supernatant fluid was better in model groups（P〈0.05）.Immunofluorescence method showed that the expression of NF-κB p65 in the cytoplasm of Caco-2 cells with hypoxia/reoxygenation injury was activated into the nu-cleus,which could be inhibited by 5%breast milk supernatant fluid boiled or unboiled,and 5%breast milk supernatant fluid was better. Conclusion Fresh breast milk can prevent intestinal epithelial cells from ischemia reperfusion injury by inhibiting the expression of NF-κB p65,which may be one of the mechanisms of breast milk preventing NEC.

关 键 词：母乳 坏死性小肠结肠炎 CACO-2细胞 NF-ΚB P65 

分 类 号：R322.74[医药卫生—人体解剖和组织胚胎学]