新生儿坏死性小肠结肠炎外周血和组织RIPK1、RIPK3的表达及其临床意义

Expression and clinical significance of RIPK1and RIPK3in peripheral blood and tissues of neonatal necrotizing enterocolitis

目的分析新生儿坏死性小肠结肠炎(NEC)外周血和组织受体相互作用蛋白激酶1(RIPK1)和RIPK3表达及其临床意义。方法选取149例NEC患儿(NEC组,Ⅰ期107例,Ⅱ+Ⅲ期42例)和同期诊治80例非NEC患儿(对照组),应用实时荧光定量PCR检测患儿外周血单个核细胞中RIPK1和RIPK3mRNA的表达。选取5例接受肠造瘘术治疗的NEC患儿的NEC组织标本,同一患儿二期关瘘时部分正常回肠组织标本为对照,Western blot检测两组组织RIPK1和RIPK3蛋白表达。比较NEC患儿外周血RIPK1和RIPK3mRNA表达与疾病严重程度的关系。多因素logistic回归分析重度NEC发生的影响因素。ROC曲线分析单独或联合检测外周血RIPK1和RIPK3mRNA表达对重度NEC的诊断效能。结果NEC组外周血RIPK1和RIPK3mRNA表达高于对照组(P<0.05)。NEC组织RIPK1和RIPK3蛋白表达高于正常回肠组织(P<0.05)。重度NEC组气腹征、多器官功能障碍综合征(MODS)发生率和外周血RIPK1、RIPK3mRNA表达高于轻度NEC组(P<0.05)。多因素logistic回归分析显示,合并MODS和气腹征、外周血RIPK1和RIPK3mRNA表达升高是重度NEC发生的独立危险因素(P<0.05)。外周血RIPK1和RIPK3 mRNA联合检测诊断重度NEC的AUC大于单独检测(P<0.05)。结论NEC患儿外周血RIPK1和RIPK3mRNA表达升高,是重度NEC发生的独立影响因素,两者联合检测对重度NEC具有较高的诊断价值。

Objective To analyze the expression and clinical significance of receptor-interacting protein kinase 1(RIPK1)and RIPK3in peripheral blood and the tissues of neonatal necrotizing enterocolitis(NEC).Methods A total of 149children with NEC(group NEC)were selected,of whom 107cases were in stageⅠ(mild NEC group)and 42cases were in stageⅡ+Ⅲ(severe NEC group).Eighty non-NEC children were taken as the controls(group C).The mRNA expressions of RIPK1and RIPK3in peripheral blood mononuclear cells were detected by real-time fluorescence quantitative PCR.Western blot was used to detect the protein expressions of RIPK1and RIPK3in NEC tissue samples from 5children undergoing enterostomy and some normal ileal tissue samples from the same child at the time of secondary closure of the fistula.The relationship between the mRNA expressions of RIPK1and RIPK3in peripheral blood of children with NEC and the severity of the disease was compared.Multivariate logistic regression analysis was used to analyze the influencing factors for severe NEC.The diagnostic efficacy of detecting RIPK1and RIPK3mRNA expressions in peripheral blood alone or in combination for severe NEC was analyzed by ROC curve analysis.Results The mRNA expressions of RIPK1and RIPK3in peripheral blood were higher in group NEC than those in group C(P<0.05).The protein expressions of RIPK1and RIPK3were higher in NEC tissue samples than those in normal ileal tissue samples(P<0.05).The incidences of pneumoperitoneum and MODS,and the expressions of RIPK1and RIPK3mRNA in peripheral blood were higher in the severe NEC group than those in the mild NEC group(P<0.05).Multivariate logistic regression analysis showed that the combination of MODS and pneumoperitoneum,and increased mRNA expressions of RIPK1and RIPK3in peripheral blood were the independent risk factors for the occurrence of severe NEC(P<0.05).The diagnostic efficacy of combined detection of RIPK1and RIPK3mRNA in peripheral blood for the diagnosis of severe NEC was higher than the single detection(P<0.05).Conclusion The increased mRNA expressions of RIPK1and RIPK3in peripheral blood of the children with NEC are independent influencing factors for the occurrence of severe NEC,and the combined detection of both has high diagnostic value for severe NEC.