摘要
目的分析新生儿坏死性小肠结肠炎(NEC)患儿外周血受体相互作用蛋白激酶3(RIPK3)、混合系列蛋白激酶样结构域(MLKL)的表达情况及其与病情严重程度的关系。方法选取92例NEC患儿纳入NEC组,并根据病情严重程度进一步分为轻度NEC组(Ⅰ级)60例和重度NEC组(Ⅱ~Ⅲ级)32例,另选取同期诊治的60例腹股沟斜疝患儿纳入对照组。采用实时荧光定量聚合酶链反应检测外周血RIPK3 mRNA、MLKL mRNA表达;采用Pearson相关分析法明确NEC组外周血RIPK3 mRNA与MLKL mRNA表达的相关性;采用免疫印迹法检测NEC回肠组织和正常回肠组织中RIPK3、MLKL蛋白表达;采用多因素Logistic回归分析明确重度NEC发生的独立危险因素。绘制受试者工作特征曲线,分析外周血RIPK3 mRNA、MLKL mRNA单独及联合预测重度NEC的价值。结果NEC组外周血RIPK3 mRNA、MLKL mRNA相对表达量分别为(2.41±0.52)、(3.03±0.64),高于对照组的(1.02±0.21)、(0.93±0.20),差异有统计学意义(P<0.001)。NEC回肠组织中RIPK3、MLKL蛋白相对灰度值分别为(1.20±0.21)、(1.13±0.24),高于正常回肠组织的(0.34±0.12)、(0.32±0.11),差异有统计学意义(P<0.05)。NEC组患儿外周血RIPK3 mRNA与MLKL mRNA相对表达量呈正相关(r=0.623,P<0.001)。重度NEC组合并气腹征、多器官功能障碍综合征、败血症者占比和RIPK3 mRNA、MLKL mRNA相对表达量均高于轻度NEC组,差异有统计学意义(P<0.05);RIPK3 mRNA、MLKL mRNA相对表达量升高是重度NEC发生的独立危险因素(P<0.05)。外周血RIPK3 mRNA、MLKL mRNA联合预测重度NEC的曲线下面积大于RIPK3 mRNA、MLKL mRNA单独预测(Z=4.127、4.261,P<0.05)。结论RIPK3 mRNA、MLKL mRNA在NEC患儿外周血中表达升高,两者均与NEC病情严重程度有关,且两者联合检测对重度NEC具有较高的预测价值。
Objective To analyze the expressions of receptor-interacting protein kinase 3(RIPK3)and mixed lineage kinase domain-like protein(MLKL)in peripheral blood of neonates with necrotizing enterocolitis(NEC)and their relationships with the severity of the disease.Methods Ninety-two children with NEC were selected and divided into NEC group,and further divided into mild group(grade Ⅰ,n=60)and severe group(grade Ⅱ to Ⅲ,n=32)according to the severity of the disease.Another 60 children with inguinal hernia were selected as control group.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expressions of RIPK3 mRNA and MLKL mRNA in peripheral blood.Pearson correlation analysis was used to determine the correlation between RIPK3 mRNA and MLKL mRNA expression in peripheral blood of the NEC group.Western blot was used to detect the expression of RIPK3 and MLKL proteins in ileal tissues of NEC and normal ileal tissues.Multivariate Logistic regression analysis was used to identify independent risk factors for severe NEC.The receiver operating characteristic curve was plotted to analyze the value of peripheral blood RIPK3 mRNA and MLKL mRNA alone and their combination for predicting severe NEC.Results The relative expression levels of RIPK3 mRNA and MLKL mRNA in peripheral blood of the NEC group were significantly higher than those in the control group[(2.41±0.52)versus(1.02±0.21),(3.03±0.64)versus(0.93±0.20),P<0.001].The relative gray values of RIPK3 and MLKL proteins in ileal tissues of NEC were significantly higher than those in normal ileal tissues[(1.20±0.21)versus(0.34±0.12),(1.13±0.24)versus(0.32±0.11),P<0.05].There was a positive correlation between the relative expression level of RIPK3 mRNA and MLKL mRNA in peripheral blood of children with NEC(r=0.623,P<0.001).The proportion of children with severe NEC complicated by pneumoperitoneum,multiple organ dysfunction syndrome,and sepsis,as well as the relative expression levels of RIPK3 mRNA and MLKL mRNA were significantly higher in the severe NEC group than in the mild NEC group(P<0.05).The relative expression levels of RIPK3 mRNA and MLKL mRNA were independent risk factors for severe NEC(P<0.05).The area under the curve for combined prediction of severe NEC by RIPK3 mRNA and MLKL mRNA in peripheral blood was larger than that for individual prediction(Z=4.127,4.261,P<0.05).Conclusion The expression levels of RIPK3 mRNA and MLKL mRNA in peripheral blood are elevated in neonates with NEC,and both are associated with the severity of NEC.The combined detection of RIPK3 mRNA and MLKL mRNA has a higher predictive value for severe NEC.
作者
黄艳
梁玉美
冯燕妮
杨松媚
HUANG Yan;LIANG Yumei;FENG Yanni;YANG Songmei(Department of Neonatology,the Affiliated Hospital of Youjiang Medical University for Nationalities,Baise,Guangxi,533000)
出处
《实用临床医药杂志》
CAS
2024年第1期62-67,共6页
Journal of Clinical Medicine in Practice
基金
广西壮族自治区卫生健康委员会科研课题(Z-M20200506)。
