儿童肺炎支原体肺炎肺泡灌洗液中受体相互作用蛋白激酶3的表达及其与细胞因子的关系研究

Expression and clinical significance of receptor-interacting protein kinase-3 levels in children with mycoplasma pneumoniae pneumonia and their relationship with inflammatory factors

目的探讨肺炎支原体肺炎(mycoplasma pneumoniae pneumonia,MPP)患儿支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中受体相互作用蛋白激酶3(receptor-interacting protein kinase-3,RIPK3)的表达水平及其与细胞因子的关系及临床意义。方法采用病例对照研究方法,选取2019年2月至2021年2月在徐州医科大学附属徐州儿童医院住院的30例难治性肺炎支原体肺炎(refractory mycoplasma pneumoniae pneumonia,RMPP)患儿作为研究1组(RMPP组),35例肺炎支原体肺炎患儿作为研究2组(MPP组),选取同期行支气管异物取出术的性别、年龄匹配的患儿20例作为对照组。采用免疫印迹法测定所有研究对象BALF中的RIPK3、混合系激酶区域样蛋白(mixed lineage kinase domain-like protein,MLKL),采用酶联免疫吸附法检测BALF中的白细胞介素(interleukin,IL)6、IL-1β及肿瘤坏死因子α(tumor necrosis factor,TNF-α)。并对组间各指标进行比较分析,Spearman秩相关分析MPP患儿BALF中RIPK3水平与MLKL、IL-6、IL-1β及TNF-α的相关性,并对RIPK3水平与MLKL水平做线性回归分析。结果 RMPP组、MPP组和对照组患儿BALF中RIPK3分别为4.85(4.33,5.68)、2.80(2.30,3.60)、1.00(1.00,1.10),MLKL分别为3.00(2.30,3.80)、1.60(1.10,2.10)、1.00(1.00,1.10),IL-6分别为(50.06±11.96)、(35.79±6.77)、(10.93±5.26) ng/L,IL-1β分别为(41.76±8.59)、(31.10±7.03)、(20.04±5.81)ng/L及TNF-α分别为(130.59±41.19)、(97.96±19.14)、(60.35±19.07) ng/L,组间比较差异均有统计学意义(Z值分别为59.609、51.164,t值分别为121.218、52.753、35.665,P均<0.001),两两比较差异均有统计学意义(P均<0.001),且均以RMPP组最高,其次为MPP组,对照组最低。MPP患儿BALF中RIPK3水平与MLKL、IL-6、IL-1β及TNF-α呈正相关(r值分别为0.711、0.676、0.725、0.651,P均<0.001),自变量RIPK3每增加1个单位,因变量MLKL增加0.432个单位(MLKL=0.432×RIPK3)。结论 RIPK3可能参与了儿童MPP的发生及发展,且与IL-6、IL-1β及TNF-α等细胞因子密切相关。

Objective To explore the level of expression,clinical significance of receptor-interacting protein kinase-3(RIPK3)in the bronchoalveolar alveolar lavage fluid(BALF)of mycoplasma pneumoniae pneumonia(MPP)in children and the relationship between RIPK3 and various cytokines.Methods Using a case-control study,30 refractory mycoplasma pneumoniae pneumonia children treated in Children′s Hospital of Xuzhou Medical University from February 2019 to February 2021 were selected as the MRPP group,35 mycoplasma pneumoniae pneumonia children as the RPP group.Meanwhile,20 sex-and age-matched hospitalized children with bronchial foreign body were selected as the case-control group.In all subjects,protein levels of RIPK3 and mixed lineage kinase domain-like protein(MLKL)were determined by Western Blot.Meanwhile,levels of IL-6,IL-1βand TNF-αwere determined by enzyme linked immuno sorbent assay(ELISA).Compare levels of different parameters between the three groups and analyze the correlation between levels of RIPK3 and MLKL,L-6,IL-1β,TNF-αin the BALF of MPP children using Spearman rank correlation analysis.Results There were statistically significant differences in the levels of RIPK3,MLKL,IL-6,IL-1βand TNF-αin BALF between RMPP group,MPP group and control group(all P<0.001).Pairwise comparisons also showed statistical differences,and the RMPP group was the highest,followed by MPP group,and the control group was the lowest.The level of RIPK3 in BALF of MPP children was positively correlated with MLKL,IL-6,IL-1βand TNF-α(r=0.711,0.676,0.725,0.651,P<0.001).Linear regression analysis shows:MLKL=0.432×RIPK3.Conclusion RIPK3 may be involved in the occurrence and development of MPP in children,and is closely related to cytokines such as IL-6,IL-1βand TNF-α.