外周血单个核细胞中AIM2炎症小体激活释放IL-18介导带状疱疹发病的机制研究*

CHEN Zehao1,2, WANG Jing2, GAO Xiangyu2, JIANG Changyu2, LI Chenguang2△, XIAO Lizu1,2△△Corresponding author: Xiao Lizu, Email: [e-mail]; Li Chenguang, Email:[e-mail]

摘要:

目的 探索在带状疱疹患者不同时期的外周血单个核细胞(peripheral blood mononuclear cell,PBMC)中AIM2炎症小体的活化和释放IL-18的情况并进行初步验证。方法 收集并分离67名AHN、58名SHN、38名PHN患者及30名健康对照者的外周血单个核细胞和血清。通过实时荧光定量PCR技术检测不同时期带状疱疹患者外周血单个核细胞中AIM2炎症小体相关基因的表达情况；蛋白质免疫印迹法检测外周血单核细胞中AIM2炎症小体蛋白的表达以及GSDMD蛋白的活化切割情况；采用蛋白质芯片技术检测血清中IL-18的分泌水平。结果实时荧光定量PCR技术检测结果发现,在基因表达水平上与健康人相比较,AIM2和IL-18的基因表达在带状疱疹AHN、SHN和PHN不同时期都有显著的上升,存在统计学差异(p < 0.05),且在PHN期差异尤为明显(p < 0.001)。蛋白质免疫印迹法检测结果发现,在蛋白质表达水平上与健康人相比较,AIM2和IL-18的基因表达在带状疱疹AHN、SHN和PHN不同时期都有显著的上升；在带状疱疹不同时期中,pro-caspase-1自我切割活化,产生有活性的caspase-1(p20),进而切割GSDMD,产生具有细胞膜成孔活性的GSDMD-NT片段,诱导细胞焦亡。蛋白质芯片技术检测结果显示不同的带状疱疹时期患者血清中IL-18的分泌水平明显的升高,尤其在PHN时期,上升最为显著。结论 外周血单个核细胞中AIM2炎症小体激活,释放的IL-18参与带状疱疹疾病的发生发展过程。

Abstract:

Objective To explore the release of IL-18 from AIM2 inflammasome activation? in peripheral blood mononuclear cells (PBMCs) of herpes zoster patients at different stages, and to conduct preliminary validation. Methods Peripheral blood mononuclear cells and serum were collected and isolated from 67 patients AHN, 58 patients with SHN, 38 patients with PHN, and 30 healthy controls. Real-time quantitative PCR was used to detect the expression of AIM2 inflammasome-related genes in peripheral blood mononuclear cells of herpes zoster patients at different stages. The expression of AIM2 inflammasome protein and the activation cleavage of GSDMD protein in peripheral blood mononuclear cells were detected using western blotting. The level of IL-18 secretion in serum was detected using protein chip technology. Results The real-time quantitative PCR results showed that the gene expression of AIM2 and IL-18 increased significantly in the different stages of herpes zoster AHN, SHN and PHN (p < 0.05), and the difference was particularly obvious in the PHN stage (p < 0.001). Western blotting results showed that the gene expression of AIM2 and IL-18 increased significantly in different stages of herpes zoster AHN, SHN and PHN compared with healthy people. During various stages of herpes zoster, pro-caspase-1 undergoes self-cleavage to produce active caspase-1 (p20). This active caspase-1 then cleaves GSDMD, resulting in the production of GSDMD-NT fragments that possess cell membrane pore-forming activity and ultimately lead to pyroptosis. The results of protein chip technology showed that the secretion level of IL-18 in the serum of patients with herpes zoster was significantly increased, particularly in the PHN stage. Conclusion The activation of the AIM2 inflammasome in peripheral blood mononuclear cells and the subsequent release of IL-18 are implicated in the onset and progression of herpes zoster disease.