Notch-芳香烃受体-白细胞介素-22通路在急性特发性血小板减少性紫癜中的作用  

The role of Notch-aryl hydrocarbon receptor-interleukin-22 pathway in idiopathic thrombocytopenic purpura

作　　者：王文焕[1] 李小燕 WANG Wenhuan;LI Xiaoyan(Department of Hematology,Bayannur Hospital,Bayannur 015000,China)

机构地区：[1]巴彦淖尔市医院血液内科

出　　处：《免疫学杂志》2020年第3期241-247,共7页Immunological Journal

摘　　要：目的观察Notch信号通路对特发性血小板减少性紫癜(idiopathic thrombocytopenic purpura,ITP)患者CD4+T淋巴细胞分泌白细胞介素(interleukin-22,IL-22)的影响,初步阐释Notch-IL-22通路在ITP发病中的作用。方法收集2015年11月至2018年1月在我科就诊的急性ITP患者31例和慢性ITP患者36例,另收集14例健康志愿者作为健康对照者。分离外周血单个核细胞(peripheral blood mononuclear cells,PBMC)和血浆,分选急性ITP患者CD4+T细胞,加入Notch信号通路抑制剂GSI刺激培养,应用实时定量PCR法检测Notch受体和Notch信号通路相关分子mRNA的相对表达量,应用流式细胞术检测CD3+CD4+IL-22+的Th22细胞比例,应用酶联免疫吸附试验检测血浆和培养上清中的IL-22水平,应用CCK-8法检测细胞增殖,应用Western blot检测Notch信号通路相关分子蛋白和磷酸化的水平。结果急性ITP患者Notch1、Notch2、Notch3 mRNA相对表达量显著高于健康对照者和慢性ITP患者(P<0.0001),但Notch受体mRNA在健康对照者和慢性ITP患者中的差异无统计学意义(P>0.05)。Th22细胞在健康对照者、急性ITP和慢性ITP患者中的比例分别为(2.00±0.15)%、(6.77±1.12)%、(2.27±0.24)%,急性和慢性ITP患者Th22细胞比例显著高于健康对照者(P<0.001),健康对照者、急性ITP和慢性ITP患者血浆IL-22的水平分别为(100.3±19.53)pg·ml^-1、(291.4±95.59)pg·ml^-1、(109.6±16.32)pg·ml^-1,急性ITP患者血浆IL-22水平显著高于健康对照者和慢性ITP患者。抑制Notch信号通路不影响CD4+T细胞增殖,但可降低急性ITP患者CD4+T细胞中Th22细胞的比例和IL-22的表达,这一过程伴随芳香烃受体(aryl hydrocarbon receptor,AhR)m RNA表达抑制和核因子-κB的磷酸化水平降低。结论Notch信号通路可通过AhR调控急性ITP患者CD4^+T细胞分泌IL-22,Notch-AhR-IL-22通路可能参与了急性ITP的发病。Idiopathic thrombocytopenic purpura(ITP),caused by the immune system mistakenly attacking and destroying platelets,is a disorder that can lead to easy or excessive bruising and bleeding.Notch signaling regulates interleukin(IL)-22 production via aryl hydrocarbon receptor(AhR) in hepatitis mouse model.Thus,the aim of current study was to investigate the effects of Notch signaling pathway on IL-22 secretion by CD4+T cells in ITP patients,and to elaborate on the role of Notch-IL-22 axis in the pathogenesis of ITP.A total of 31 acute ITP patients and 36 chronic ITP patients who were hospitalized in our department from November 2015 to January 2018 were enrolled in this study.Fourteen healthy individuals matched with the patients in age and sex ratio were enrolled as healthy control(HC).Peripheral blood mononuclear cells(PBMC) and plasma were isolated from all enrolled subjects.CD4+T cells,which were purified from acute ITP patients,were stimulated with Notch signaling inhibitor GSI.Notch receptors and Notch signaling related molecules mRNA levels were measured by real-time PCR;the percentage of CD3+CD4+IL-22+Th22 cells was investigated by flow cytometry;IL-22 concentration in plasma and cultured supernatants were assessed by enzyme linked immunosorbent assay.Cellular proliferation was analyzed by cell counting kit-8.Notch signaling related molecules expression and phosphorylation were investigated by Western blot.Data showed that Notch1,Notch2,Notch3 mRNA relative levels were significantly elevated in acute ITP patients when compared with HC(P<0.000 1).However,there were no remarkable differences of Notch receptors between HC and chronic ITP patients(P>0.05).The percentage of Th22 in HC,acute ITP,and chronic ITP were(2.00±0.15)%,(6.77±1.12)%,and(2.27±0.24)%,respectively.Th22 frequency in ITP was robustly higher than that in HC.Plasma IL-22 concentration in HC,acute ITP,and chronic ITP were(100.3±19.53) pg·ml^-1,(291.4±95.59) pg·ml^-1,and(109.6±16.32) pg·ml^-1,respectively.IL-22 expression in acute ITP was si

关 键 词：特发性血小板减少性紫癜 NOTCH信号通路 白细胞介素-22 免疫调控 

分 类 号：R512.6[医药卫生—内科学]