β-catenin通过ASK1抑制铜绿假单胞菌引起的炎性反应  

β-catenin inhibited Pseudomonas aeruginosa-induced inflammation via suppressing ASK1 expression

作　　者：傅强[1] 王伟佳[1] 黄福达[1] 缪丽韶[1] 杨山虹[1] 陈康[1] FU Qiang;WANG Weijia;HUANG Fuda;MIAO Lishao;YANG Shanhong;CHEN Kang(Center of Laboratory Medicine,Peoples′Hospital of Zhongshan City,Zhongshan,Guangdong 528403,China)

机构地区：[1]中山市人民医院检验医学中心,广东中山528403

出　　处：《国际检验医学杂志》2021年第14期1679-1682,共4页International Journal of Laboratory Medicine

基　　金：国家自然科学基金项目(81401645);广东省中山市社会公益科技研究项目(2018B1032)。

摘　　要：目的探讨β-catenin抑制铜绿假单胞菌感染引起的炎性反应的机制。方法采用普通聚合酶联反应(PCR)的方法和Western-blot的方法在过表达β-catenin的RAW264.7细胞和对照细胞中验证差异表达细胞凋亡信号调节激酶1(ASK1)转录水平和蛋白的表达;应用小干扰RNA(siRNA)沉默ASK1,在铜绿假单胞菌感染的RAW264.7细胞中,检测炎症因子白细胞介素-6(IL-6)和IL-1β的表达;构建铜绿假单胞菌角膜炎的动物模型,应用siRNA沉默ASK1,观察ASK1在体内对炎性反应的调控;应用ASK1质粒,在过表达β-catenin的RAW264.7细胞中,采用实时荧光定量PCR(Real-time PCR)检测细胞因子IL-6和IL-1β的基因水平,观察ASK1对β-catenin介导的炎性反应的回复作用。结果在过表达β-catenin的RAW264.7细胞中ASK1的转录水平和蛋白水平明显降低。在感染铜绿假单胞菌后,沉默ASK1可抑制炎症因子IL-6和IL-1β的表达;构建铜绿假单胞菌角膜炎的动物模型,实验结果发现沉默ASK1后临床评分降低,角膜炎症程度和范围降低,同时可抑制角膜中IL-6和IL-1β的表达。进一步机制探讨发现ASK1可减弱β-catenin对炎症的抑制作用。结论β-catenin通过调控ASK1抑制铜绿假单胞菌感染引起的炎性反应。Objective To explore the underlying mechanism ofβ-catenin in Pseudomonas aeruginosa-induced inflammation.Methods The mRNA levels and protein levels of apoptosis signal regulated kinase 1(ASK1)were detected by PCR and Western-blot inβ-catenin overexpressing RAW264.7 cells and control cells respectively.The expression of IL-6 and IL-1βwere detected by Real-time PCR in Pseudomonas aeruginosa infected RAW264.7 cells after silencing ASK1 by using siRNA.Pseudomonas aeruginosa keratitis were constructed,and ASK1 was silenced by siRNA to observe the regulation of ASK1 on inflammatory response in vivo.The expression of IL-6 and IL-1βwere detected by Real-time PCR in overexpressing RAW264.7 cells,followed by ASK1 vector application.Results The mRNA levels and protein levels of ASK1 were down-regulated inβ-catenin overexpressing RAW264.7 cells.The expression of IL-6 and IL-1βwere down-regulated in Pseudomonas aeruginosa infected RAW264.7 cells after silencing ASK1.Pseudomonas aeruginosa keratitis were constructed,and the results showed that ASK1 silencing could reduce the clinical score,the degree and scope of keratitis,and inhibit the expression of IL-6 and IL-1βin cornea.Further data showed that ASK1 could reduce the suppression of inflammatory cytokine regulated byβ-catenin overexpression.Conclusionβ-catenin inhibited Pseudomonas aeruginosa-induced inflammation via suppressing ASK1 expression.

关 键 词：Β-CATENIN 铜绿假单胞菌 细胞凋亡信号调节激酶1 炎性反应 

分 类 号：R440[医药卫生—诊断学]