钙敏感受体介导的NLRP3炎症小体信号通路在脓毒症急性肺损伤患者中的作用机制

Mechanism of Calcium-sensing Receptor-mediated NLRP3 Inflammasome Signaling Pathway in Patients with Sepsis and Acute Lung Injury

脓毒症(sepsis)是一种由细菌、病毒、真菌等微生物感染引起的全身性炎症反应综合征,其发病机制十分复杂.钙敏感受体位于细胞膜上与免疫细胞的活化和炎症反应密切相关.为了探讨钙敏感受体介导的NLRP3炎症小体信号通路在脓毒症急性肺损伤患者中的表达变化机制.选取2022年3月至2023年3月我院收治的脓毒性急性肺损伤患者110例为研究对象,同期普通脓毒症患者55例作为对照组.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.q RT-PCR法和Western blot比较了不同组别中外周血单核细胞(peripheral blood mononuclear cells,PBMCs)中NLRP3、Caspase-1的m RNA和蛋白表达水平.酶联免疫吸附法检测外周血中钙敏感受体以及IL-1β、IL-18和TNF-α水平在不同组间的表达水平.结果表明:高危组PBMCs钙敏感受体、NLRP3和Caspase-1的m RNA和蛋白表达水平明显高于中危组、低危组及普通脓毒症患者组(P<0.05);而中危组上述指标显著高于低危组及普通脓毒症患者组(P<0.05).高危组血清IL-1β、IL-18、TNF-α表达水平明显高于中危组、低危组及普通脓毒症患者组;而中危组明显高于低危组及普通脓毒症患者组(P<0.05).Pearson相关性分析发现钙敏感受体的水平与肺损伤的严重程度和病死率均呈正相关(P<0.05).通过ROC曲线对脓毒症急性肺损伤患者病情严重程度的诊断价值,钙敏感受体对脓毒症患者发生急性肺损伤诊断曲线下面积为0.9336(95%CI:0.8988-0.9685),灵敏度、特异度分别为78.50%、68.30%;显著优于LIPS和APACHEⅡ评分.钙敏感受体介导的NLRP3炎症小体信号通路参与了脓毒症急性肺损伤患者的机体炎症反应,对于患者病情严重程度及预后评估具有应用价值.

Sepsis is a systemic inflammatory response syndrome caused by microbial infections such as bacteria,viruses,and fungi,The pathogenesis of sepsis is very complex.Calcium sensitive receptors are located on the cell membrane and closely related to immune cell activation and inflammatory response.In order to explore the expression mechanism of NLRP3 inflammasome signaling pathway mediated by calcium sensitive receptors in patients with sepsis acute lung injury,110 patients with sepsis acute lung injury admitted to our hospital from March 2022 to March 2023 were selected as the research subjects,55 patients with common sepsis during the same period served as the control group.Peripheral venous blood samples were collected from all patients within 24 hours of admission,while the control group collected peripheral venous blood on the day of physical examination.QRT-PCR and Western blot were used to compare the mRNA and protein expression levels of NLRP3 and Caspase-1 in PBMCs from different groups.Enzyme linked immunosorbent assay was used to detect calcium sensitive receptors and IL-1β,IL-18 and TNF-αin peripheral blood.The results showed that the mRNA and protein expression levels of calcium sensitive receptors,NLRP3,and Caspase-1 in PBMCs in the high-risk group were significantly higher than those in the medium risk group,low risk group,and ordinary sepsis patient group(P<0.05).The above indicators in the medium risk group were significantly higher than those in the low risk group and the ordinary sepsis patient group(P<0.05).The expression levels of IL-1β、IL-18,TNF-αin the serum in the high risk group was significantly higher than that of the medium risk group,low risk group,and ordinary sepsis patient group;The mdium risk group was significantly higher than the low risk group and the group of ordinary sepsis patients(P<0.05).Pearson correlation analysis found that the level of calcium sensitive receptors was positively correlated with the severity and mortality of lung injury(P<0.05).The diagnostic value of the ROC curve for the severity of acute lung injury in patients with sepsis,the area under the curve of calcium-sensitive receptors for the diagnosis of acute lung injury in sepsis patients is 0.9336(95%CI:0.8988-0.9685)and the sensitivity and specificity were 78.50%and 68.30%,respectively,significantly superior to LIPS and APACHE II scores.The NLRP3 inflammasome signaling pathway mediated by calciumsensitive receptors is involved in the inflammatory response of patients with sepsis and acute lung injury and has practical value in evaluating the severity and prognosis of the patient's condition.