脓毒症急性肺损伤患者RhoA/ROCK1和NF-κB/NLRP3信号通路及其下游信号分子的变化

Changes of RhoA/ROCK1 and NF-κB/NLRP3 signaling pathways and their downstream signaling molecules in patients with sepsis and acute lung injury

目的 分析Ras同源基因家族成员A(RhoA)/Rho激酶1(ROCK1)、核因子κB(NF-κB)/核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)信号通路及其下游信号分子在脓毒症患者急性肺损伤(ALI)中的变化。方法 回顾性分析海口市第四人民医院2018年1月-2020年12月122例脓毒症患者临床资料,其中41例并发ALI(ALI组),81例无ALI(非ALI组),并纳入32名健康体检者作为对照组。脓毒症患者均于入院24 h内收集外周静脉血标本,对照组则收集体检当日外周静脉血。比较3组外周血单个核细胞(PBMCs)NF-κB、NLRP3信使核糖核酸(mRNA)表达水平,并记录NF-κB/NLRP3信号通路上游基因(RhoA、ROCK1)、下游基因[半胱氨酸天冬氨酸特异性蛋白酶-1(caspase-1)]mRNA表达水平及下游炎症因子[白细胞介素-1β(IL-1β)、IL-18];根据肺损伤预测评分(LIPS)将41例并发ALI的脓毒症患者分为LIPS≥7分组,LIPS<7分组,比较两组上述基因mRNA、炎症因子水平,并评估脓毒症并发ALI患者LIPS评分与上述基因mRNA、炎症因子表达水平的相关性。结果 ALI组LIPS评分高于非ALI组(P<0.05)。ALI组与非ALI组PBMCs NF-κB、NLRP3、RhoA、ROCK1、caspase-1 mRNA相对表达水平及血清IL-1β、IL-18高于对照组(P<0.05),ALI组高于非ALI组(P<0.05)。脓毒症并发ALI患者中,LIPS≥7分组PBMCs NF-κB、NLRP3、RhoA、ROCK1、caspase-1 mRNA相对表达水平及血清IL-1β、IL-18高于LIPS<7分组(P<0.05);Pearson相关性分析显示,LIPS评分与上述各基因mRNA及炎症因子均呈正相关(r=0.389、0.472、0.511、0.539、0.518、0.469、0.457,P均<0.05)。结论 Rho/ROCK、NF-κB/NLRP3信号通路及其下游信号分子可能共同参与了脓毒症ALI的发生发展,临床可针对该发生机制探寻治疗新方向。

OBJECTIVE To analyze the changes of Ras homologous gene family member A(RhoA)/Rho kinase 1(ROCK1) and nuclear factor-κB(NF-κB)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) signaling pathways and their downstream signaling molecules in patients with sepsis complicated with acute lung injury(ALI). METHODS The clinical data of 122 patients with sepsis in the Fourth People’s Hospital of Haikou Jan. 2018 to Dec. 2020 were retrospectively analyzed. Among them, 41 cases were complicated with ALI(ALI group) and 81 cases were without ALI(non-ALI group), and 32 healthy subjects were included as control group. Peripheral venous blood specimens were collected among patients with sepsis within 24 h of admission, and peripheral venous blood were collected in control group on the day of physical examination. The expression levels of NF-κB and NLRP3 messenger ribonucleic acid(mRNA) in peripheral blood mononuclear cells(PBMCs) were compared among the three groups, and the mRNA expression levels of NF-κB/NLRP3 signaling pathway upstream genes(RhoA, ROCK1) and downstream genes [cysteine aspartate-specific protease-1(caspase-1)] and levels of downstream inflammatory factors [interleukin-1β(IL-1β), IL-18] were recorded. According to the lung injury prediction score(LIPS), 41 patients with sepsis complicated with ALI were divided into LIPS ≥ 7 points group and LIPS <7 points group, and the mRNA expression levels of the above-mentioned genes and levels of inflammatory factors were compared between the two groups, and the correlation between LIPS score and expression levels of the above-mentioned gene mRNA and inflammatory factors were evaluated among patients with sepsis and ALI. RESULTS The LIPS score of ALI group was higher than that of non-ALI group(P<0.05). The mRNA relative expression levels of NF-κB, NLRP3, RhoA, ROCK1 and caspase-1 in PBMCs and levels of serum IL-1β and IL-18 in ALI group and non-ALI group were higher than those in control group(P<0.05), and the above levels in ALI group were higher than those in non-ALI group(P<0.05). Among patients with sepsis complicated with ALI, the mRNA relative expression levels of NF-κB, NLRP3, RhoA, ROCK1 and caspase-1 in PBMCs and levels of serum IL-1β and IL-18 of LIPS≥7 points group were higher than those of LIPS<7 points group(P<0.05). Pearson correlation analysis showed that the LIPS score was positively correlated with the mRNA expression levels of the above-mentioned genes and levels of inflammatory factors(r=0.389, 0.472, 0.511, 0.539, 0.518, 0.469, 0.457, all P<0.05). CONCLUSION Rho/ROCK and NF-κB/NLRP3 signaling pathways and their downstream signaling molecules may jointly participate in the occurrence and development of sepsis with ALI, and new treatment directions can be explored for this mechanism.