RIP1-RIP3-MLKL信号通路在急性冠状动脉综合征病人外周血单核细胞中的表达

Expression of RIP1-RIP3-MLKL signaling pathway in peripheral blood monocytes of patients with acute coronary syndrome

目的:研究急性冠状动脉综合征(ACS)病人外周血单核细胞(PBMCs)上刺激受体相互作用蛋白1(RIP1)-RIP3-混合谱系激酶结构域样假激酶(MLKL)信号通路的表达变化,并分析其可能机制。方法:选取初诊为ACS病人60例,其中不稳定型心绞痛(UAP)病人31例,急性心肌梗死(AMI)病人20例,对照组选取同期健康体检者19名。采集所有受试者外周静脉血,分别提取血浆和外周血单核细胞(PBMCs),酶联免疫吸附实验(ELISA)和比浊法分别检测血浆中白细胞介素(IL)-1和IL-18、MLKL的水平,运用Real Time-PCR检测MLKL的mRNA表达,免疫印迹法检测PBMCs中RIP1、RIP3和MLKL的蛋白表达。结果:3组受试者血浆IL-1、IL-18含量和MLKL含量差异均有统计学意义(P<0.01),PBMCs中RIP1和RIP3蛋白表达增加(P<0.05),MLKL的mRNA和蛋白表达亦增加(P<0.05)。与UAP组比较,AMI组病人外周血浆中IL-1、IL-18和MLKL进一步升高,RIP1和RIP3的蛋白表达的进一步增加(P<0.05),MLKL的mRNA和蛋白表达增高。结论:UAP和AMI病人体内存在不同程度的炎症活化,RIP1-RIP3-MLKL信号通路在PBMCs中的表达量随病情的严重程度逐渐增加,提示RIP1-RIP3-MLKL信号通路可能在ACS的不同类型发病中起重要作用。

Objective:To investigate the expression changes of RIP1-RIP3-MLKL signaling pathway in peripheral blood mononuclear cells(PBMCs)of patients with acute coronary syndrome(ACS)and analyze the mechanism.Methods:Sixty patients with ACS were enrolled,including 31 patients with unstable angina(UAP)and 20 patients with acute myocardial infarction(AMI).The control group received 19 healthy cases.Peripheral venous blood was collected from all subjects,plasma and monocytes were extracted,and interleukin(IL)-1 and IL-18 were detected by enzyme-linked immunosorbent assay(ELISA)and turbidimetry.The level of MLKL was detected by real-time PCR.The expression of RIP1,RIP3 and MLKL in PBMCs were detected by Western blotting.Results:The levels of IL-1,IL-18 and MLKL in plasma of 3 groups were significantly different(P<0.01),RIP1 and RIP3 protein expression increased in PBMCs(P<0.05),mRNA and protein expression of MLKL were also increased(P<0.05).Compared with the UAP group,IL-1,IL-18 and MLKL were increased in peripheral blood of patients with AMI,and the protein expression of RIP1 and RIP3 were increased(P<0.05).The expressions of MLKL mRNA and protein expression were also increased.Conclusions:There are different degrees of inflammatory activation in UAP and AMI patients.The expression of RIP1-RIP3-MLKL signaling pathway in PBMCs gradually increases with the severity of the disease,suggesting that RIP1-RIP3-MLKL signaling pathway may play an important role in ACS.