白介素-18及NOD样受体蛋白3炎性小体在痤疮发病中的作用研究  

IL-18 and NLRP3 in the Process of Acnevulgaris

作　　者：李晓娟 林新瑜[1] 沈柱 邓秋 刘颖[1] 程石[1] 刘伟[1] LI Xiao-juan;LIN Xin-yu;SHEN Zhu;DENG Qiu;LIU Ying;CHENG Shi;LIU Wei(Department of Dermatology and Venereology,Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital,Chengdu 610072,China)

机构地区：[1]四川省医学科学院.四川省人民医院皮肤病性病研究所,成都610072

出　　处：《四川大学学报（医学版）》2019年第1期66-70,共5页Journal of Sichuan University(Medical Sciences)

基　　金：2015年四川省医学科学院.四川省人民医院博士基金(No.2015BS03)资助

摘　　要：目的探讨白介素-18(IL-18)及NOD样受体蛋白3(NLRP3)炎性小体在痤疮炎性发病机制中的作用。方法以痤疮丙酸杆菌悬液[感染复数(MOI)=0、10、20、30)]刺激人角质形成细胞(NHEK)12h、24h、36h,ELISA法检测NHEK中IL-18蛋白的表达量,real-time PCR检测NHEK中IL-18mRNA的表达。将NHEK细胞分为3组:特异性NLRP3小干扰RNA(siRNA)转染NHEK 36h后,以MOI=30的痤疮丙酸杆菌悬液刺激NHEK 36h(siRNA组);不转染、不以菌液刺激的NHEK为空白对照组;不转染、只以MOI=30的痤疮丙酸杆菌悬液刺激36h的NHEK为阳性对照组。ELISA法和real time-PCR法检测3组NHEK中IL-18蛋白和基因的表达量,Western blot法检测NLRP3炎性小体的表达情况。结果 NHEK经痤疮丙酸杆菌悬液刺激后IL-18蛋白和基因的表达量增加,具有时间相关性和剂量相关性(r均>0.75,P<0.05),以菌悬液MOI=30、刺激36h最为明显。siRNA组IL-18蛋白和基因表达量、NLRP3炎性小体的表达量较阳性对照组降低,但高于空白对照组,差异有统计学意义(P<0.05)。结论痤疮丙酸杆菌刺激NHEK分泌IL-18可能需要NLRP3炎性小体的参与。Objective To determine the role of inlerleukin-18(IL-18)and nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3(NLRP3)in acne vulgaris.Methods We used propionibacterium acnes(P.acnes)suspensions[multiplicity of infection(MOI)=0,10,20,30]to stimulate normal human epidermal kerationocytes(NHEKs)for 12,24,and 36 h,respectively.Enzyme-linked immunosorbent assay(ELISA)was performed to detect the protein level of IL-18.Real-time quantitative PCR(real-time PCR)was adopted to detect the mRNA of IL-18.The NHEKs were divided into three groups:(1) siRNA group:NHEKs were pretreated with siRNA for 36 h,followed by 36 hexposure to MOI=30of P.ances suspensions;(2) blank control group:NHEKs free from siRNA transfection and P.ances suspensions;(3) positive control group:NHEKs free from siRNA transfection were exposed to P.ances suspensions(MOI=30).The expression of NLRP3 was detected by Western blot.Results The expressions of protein and mRNA of IL-18 increased with exposure to P.ances suspensions in a dose responsive way(r >0.75,P<0.05),with the peak effects showing for MOI=30at 36 h.The expression of IL-18 decreased in the siRNA group compared with the positive control,but was still higher than that of the blank group(P<0.05).Conclusion P.ances stimulates NHEK cells to secrete IL-18.The process possibly requires the involvement of NLRP3.

关 键 词：痤疮丙酸杆菌 IL-18 NLRP3炎性小体 

分 类 号：R758.733[医药卫生—皮肤病学与性病学]