Luo Y, Huang Y Y, Jin Y, Li C R, Yang J
Rheumatology & Immunology Department of Shenzhen Children's Hospital, Shenzhen 518038, China.
Zhonghua Er Ke Za Zhi. 2018 Dec 2;56(12):939-944. doi: 10.3760/cma.j.issn.0578-1310.2018.12.010.
To investigate the effect of microRNAs (miR)-21 on the expression of interleukin (IL)-10 in B cell of patients with Henoch-Schonlein purpura (HSP). From March 2016 to January 2017, twenty-four children with HSP hospitalized in rheumatology and immunology department of Shenzhen Children's Hospital were enrolled into the study, including 12 males and 12 females. Patients were divided into purpura nephritis group (HSPN, 14 cases) and non-nephritis group (NHSPN, 10 cases). The age-matched 34 healthy children were included as the control group for prospective cohort study. The expression levels of IL-10 in peripheral blood B cells (CD19(+)), transitional B cells (CD19(+) CD24(hi)CD38(hi)) and naïve B cells (CD19(+)CD24(int)CD38(int)) from patients with HSP and healthy children were detected by flow cytometry (FCM). Expression of microRNAs related to IL-10 in 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 control-FAM and agomir-21-5p-FAM were transfected 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-10 was detected by FCM after transfection. For quantitative data of normal distribution, test was used for two samples comparison and multiple comparisons among three groups were conducted by ANOVA. Spearman test was used for correlation analysis. (1) The CD19(+) B cells and its two populations at different differentiation stages all could express IL-10. The expression levels of IL-10 in three B cell populations in patients were significantly lower than those in healthy controls (1.4±0.2 2.4±0.3, 3.501, 0.01; 1.2±0.2 2.2±0.3, 2.688, 0.05; 1.6±0.3 2.7±0.4, 2.498, 0.05). Compared with healthy control and NHSPN groups, the expression of IL-10 in CD19(+) B cells from patients within HSPN group was the lowest, and the difference was statistically significant (1.1±0.2 2.4±0.3, 1.8±0.3, 4.006, 2.362, 0.001, 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±0.9 . 3.5±2.8, 2.962, 0.01). There was no significant change in the other microRNAs. (3) The expression of IL-10 was positively correlated with the expression of miR-21-5p in the B cells of patients with HSP (0.778, 0.001). (4) The expression of IL-10 in B cells of miR-21-5p group was significantly higher than that in negative control group (2.7±0.2 1.6±0.3, 3.091, 0.05). 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.
探讨微小RNA（miR）-21对过敏性紫癜（HSP）患者B细胞中白细胞介素（IL）-10表达的影响。2016年3月至2017年1月，选取深圳市儿童医院风湿免疫科收治的24例HSP患儿纳入研究，其中男12例，女12例。将患者分为紫癜性肾炎组（HSPN，14例）和非肾炎组（NHSPN，10例）。选取34例年龄匹配的健康儿童作为前瞻性队列研究的对照组。采用流式细胞术（FCM）检测HSP患者和健康儿童外周血B细胞（CD19(+)）、过渡性B细胞（CD19(+)CD24(hi)CD38(hi)）和初始B细胞（CD19(+)CD24(int)CD38(int)）中IL-10的表达水平。采用实时荧光定量PCR法检测B细胞中与IL-10相关的微小RNA表达，包括miR-21-5p、miR-106a-5p、miR-98-3p、miR-142-3p、miR-142-5p、miR-98-5p、miR-155-5p和miR-let7b-5p。将agomir阴性对照-FAM和agomir-21-5p-FAM转染至HSP患者的B细胞中。通过激光扫描共聚焦显微镜和FCM观察B细胞对miRNA的摄取情况，并在转染后采用FCM检测IL-10的表达。对于正态分布的定量数据，两组比较采用t检验，三组间多重比较采用方差分析。采用Spearman检验进行相关性分析。（1）CD19(+)B细胞及其不同分化阶段的两个亚群均能表达IL-10。患者三个B细胞亚群中IL-10的表达水平均显著低于健康对照组（1.4±0.2对2.4±0.3，3.501，0.01；1.2±0.2对2.2±0.3，2.688，0.05；1.6±0.3对2.7±0.4，2.498，0.05）。与健康对照组和NHSPN组相比，HSPN组患者CD19(+)B细胞中IL-10的表达最低，差异有统计学意义（1.1±0.2对2.4±0.3，1.8±0.3，4.006，2.362，0.001，0.05）。（2）HSPN患者B细胞中miR-21-5p的表达低于健康对照组（1.2±0.9对3.5±2.8，2.962，0.01）。其他微小RNA无明显变化。（3）HSP患者B细胞中IL-10的表达与miR-21-5p的表达呈正相关（0.778，0.001）。（4）miR-21-5p组B细胞中IL-10的表达显著高于阴性对照组（2.7±0.2对1.6±0.3，3.091，0.05）。HSP患者外周血B细胞中miR-21-5p表达不足是B细胞中IL-10表达降低的原因之一。
