摘要
目的探讨微小RNA(miRNA)一21对过敏性紫癜患儿外周血B细胞白细胞介素(IL)10表达的影响。方法前瞻性队列研究。选取2016年3月至2017年1月深圳市儿童医院风湿免疫科住院过敏性紫癜患儿24例(男女各12例)为研究对象,分为紫癜肾炎组(14例)和非。肾炎组(10例);选取同年龄健康儿童34名为对照组。流式细胞术检测过敏性紫癜患儿及对照组儿童外周血B细胞(CDl9^+),过渡性B细胞(CDl9^+CD24^hiCD38^hi)和初始B细胞(CDl9^+CD24^intCD38^int)胞内IL一10表达水平。实时PCR检测B细胞与IL-10表达相关的miRNA:miR-21-5p、miR-106a-5p、miR一98—3p、miR-142-3p、miR-142—5p、miR一98—5p、miR-155—5p、miR-1et7b-5p。转染阴性对照(agomir NC—FAM)和agomir一21—5p—FAM至过敏性紫癜患儿外周血B细胞,激光共聚焦显微镜及流式观察miRNA在B细胞的摄取,并比较转染后B细胞IL.10的表达。组间比较采用t检验或单因素方差分析,相关性分析采用Spearman检验。结果(1)CDl9^+B细胞及其两种不同分化阶段的B细胞群体均可表达IL一10,过敏性紫癜患儿三种B细胞群体(CDl9^+、CDl9^+CD24^hiCD38^hi、CDl9^+CD24^intCD38^int)IL-10表达均明显低于健康对照组(1.4+0.2比2.4±0.3,t=3.501,P<0.01;1.2±0.2比2.2±0.3,t=2.688,P<0.05;1.6±0.3比2.7±O.4,t=2.498,P<0.05);紫癜肾炎组与健康对照及非肾炎组比较,B细胞IL一10表达最低,两两相比差异均有统计学意义(1.1±0.2比2.4±0.3,1.8±0.3,t=4.006,2.362,P<0.001,P<O.05)。(2)过敏性紫癜患儿B细胞miR一21—5p表达低于健康对照组,紫癜肾炎组更显著,差异有统计学意义(1.2±0.9比3.5±2.8,t=2.962,P<0.01);其余miRNA表达水平无明显改变。(3)相关性分析显示过敏性紫癜B细胞IL一10与miR.21.5p表达呈正相关(r=0.778,P<0.001)。(4)与阴性对照组相比,转染agomir一21—5p可明显增强过敏性紫癜患儿B细胞IL一10表达,差异有统计学意义(2.7±0.2比1.6±0.3,t=3.091,P<0.05)。结论过敏性紫癜患儿外周血B细胞miR一21—5p表达不足,是引起B细胞IL一10表达降低的原因之一。
Objective To investigate the effect of microRNAs (miR)-21on the expression of interleukin (IL)-10in B cell of patients with Henoch-Sehonlein purpura (HSP).Methods From March 2016to January 2017,twenty-four children with HSP hospitalized in rheumatology and immunology department of Shenzhen Children's Hospital were enrolled into the study,including 12males and 12females. Patients were divided into purpura nephritis group (HSPN,14cases)and non-nephritis group (NHSPN, 10cases).The age-matched 34healthy children were included as the control group for prospective cohort study.The expression levels of IL-10in peripheral blood B cells (CD19+),transitional B cells (CD19^+CD24^hiCD38^hi)and naive B cells (CD19+CD24m^intCD38^int)from patients with HSP and healthy children were detected by flow eytometry (FCM).Expression of microRNAs related to IL-10in B cells were quantitated by real-time PCR,including miR-21-5p,miR-106a-5p,miR-98-3p,miR-142-3p,miR-142-5p,miR-98-5p, miR-155-5p and miR-let7b-5p.Agomir negative eontrol-FAM and agomir-21-5p-FAM were transfeeted into B cells from patients with HSP.The uptake of miRNA by B cells was observed by laser scanning confocal microscope and FCM,and the expression of IL-10was detected by FCM after transfection.For quantitative data of normal distribution,t test was used for two samples comparison and multiple comparisons among three groups were conducted by ANOVA.Spearman test was used for correlation analysis.Results (1)The CD19^+B cells and its two populations at different differentiation stages all could express IL-10. The expression levels of IL-10in three B cell populations in patients were significantly lower than those in healthy controls (1.4±0.2vs.2.4±0.3,t=3.501,P<0.01;1.2±0.2vs.2.2±0.3,t=2.688,P<0.05;1.6±0.3vs. 2.7±0.4,t=2.498,P<0.05).Compared with healthy control and NHSPN groups,the expression of IL-10in CD19^+B cells from patients within HSPN group was the lowest,and the differenec was statistically significant (1.1±0.2vs.2.4±0.3,1.8±0.3,t=4.006,2.362,P<0.001,P<0.05).(2)The expression of miR-21-5p in B cell in patients with HSPN was lower than that in healthy control group (1.2±0.9vs.3.5±2.8,t=2.962, P<0.01).There was no significant change in the other microRNAs.(3)The expression of IL-10was positively correlated with the expression of miR-21-5p in the B cells of patients with HSP (r=0.778,P< 0.001).(4)The expression of IL-10in B cells of miR-21-5p group was significantly higher than that in negative control group (2.7±0.2vs.1.6±0.3,t=3.091,P<0.05)'.Conclusion The insufficient expression of miR-21-5p in peripheral blood B cells of patients with HSP is one of the reasons for the reduction of IL-10 expression in B cells.
作者
罗颖
黄艳艳
金言
李成荣
杨军
Luo Ying;Huang Yanyan;Jin }Tan;Li Chengrong;Yang Jun(Rheumatology &Immunology Department of Shenzhen Children's Hospital,Shenzhen 518038,China)
出处
《中华儿科杂志》
CAS
CSCD
北大核心
2018年第12期939-944,共6页
Chinese Journal of Pediatrics
