病毒性脑炎病儿脑脊液中微小RNA-21表达水平与辅助性T细胞17/调节性T细胞平衡的相关性分析

Expression of miR-21 in cerebrospinal fluid of children with viral encephalitis and its correla‑tion with Th17/Treg balance

目的探讨病毒性脑炎病儿脑脊液中微小RNA-21(miR-21)表达水平与辅助性T细胞17(Th17)/调节性T细胞(Treg)平衡的相关性。方法选取2017年9月至2019年7月在抚顺市中心医院就诊的病毒性脑炎病儿(观察组)108例作为研究对象,其中,轻症病儿(轻症组)57例,重症病儿(重症组)51例,选取同期抚顺市中心医院患病毒性脑炎后已康复的儿童(对照组)110例作为对照组。采集受试者脑脊液,实时荧光定量聚合酶链式反应(qRT-PCR)法检测miR-21相对表达量;酶联免疫(ELI‐SA)法检测白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-17(IL-17)、转化生长因子-β(TGF-β)水平;流式细胞仪分析Treg、Th17细胞水平;Pearson法分析病毒性脑炎病儿脑脊液中miR-21、Th17/Treg水平与IL-6、IL-10、IL-17、TGF-β水平,miR21水平与Th17/Treg水平的相关性。结果与对照组比较,观察组脑脊液中miR-21[(3.51±0.74)比(2.27±0.59)]、Th17[(3.14±0.62)%比(2.38±0.37)%]、Th17/Treg[(3.16±0.97)比(1.89±0.55)]、IL-6[(29.63±5.32)ng/L比(22.84±4.36)ng/L]、IL-17[(25.69±4.79)ng/L比(19.44±3.36)ng/L]水平明显升高(P<0.05),Treg[(1.04±0.23)%比(1.37±0.36)%]、IL-10[(29.68±5.22)ng/L比(36.87±6.63)ng/L]、TGF-β[(634.14±63.58)ng/L比(771.62±95.74)ng/L]水平明显降低(P<0.05);与轻症组相比,重症组病毒性脑炎病儿脑脊液中miR-21、Th17、Th17/Treg、IL-6、IL-17水平明显升高(P<0.05),Treg、IL-10、TGF-β水平明显降低(P<0.05);病毒性脑炎病儿脑脊液中miR-21水平、Th17/Treg水平与IL-6、IL-17水平,miR-21水平与Th17/Treg均明显正相关(P<0.05),miR21水平、Th17/Treg水平与IL-10、TGF-β水平均明显负相关(P<0.05)。结论miR-21表达水平增加与Th17/Treg细胞平衡失衡密切相关,这一相关过程可能有IL-6、IL-10、IL-17、TGF-β的参与,可能均参与了病毒性脑炎的发病及病情加重过程。

Objective To study the expression level of microRNA-21(miR-21)and the balance of helper T cell 17(Th17)/regulatory T cell(Treg)in cerebrospinal fluid of children with viral encephalitis,and to explore the correlation between the expression level of miR-21 and the balance of Th17/Treg.Methods One hundred and eight children with viral encephalitis(observation group)in Depart‐ment of Pediatrics,Fushun Central Hospital from September 2017 to July 2019 were selected as the study objects,including 57 chil‐dren with mild illness(light group)and 51 children with severe illness(severe group).110 children(control group)who had recovered from viral encephalitis in the same period were selected as the control.The cerebrospinal fluid was collected and the relative expression of miR-21 was detected by real-time fluorescence quantitative PCR(qRT-PCR)method;the levels of interleukin(IL)-6,IL-10,IL-17 and transforming growth factor-β(TGF-β)were detected by enzyme-linked immunoassay(ELISA);flow cytometry was used to analyze Treg and Th17 cell levels;and Pearson's method was used to analyze the correlations between the levels of miR-21,Th17/Treg and the levels of IL-6,IL-10,IL-17,TGF-β,and the correlation between levels of miR-21 and Th17/Treg.Results The levels of miR-21[(3.51±0.74)vs.(2.27±0.59)],Th17[(3.14±0.62)%vs.(2.38±0.37)%],Th17/Treg[(3.16±0.97)vs.(1.89±0.55)],IL-6[(29.63±5.32)ng/L vs.(22.84±4.36)ng/L]and IL-17[(25.69±4.79)ng/L vs.(19.44±3.36)ng/L]in cerebrospinal fluid of the observation group were signifi‐cantly higher than those of the control group(P<0.05),while the levels of Treg[(1.04±0.23)%vs.(1.37±0.36)%],IL-10[(29.68±5.22)ng/L vs.(36.87±6.63)ng/L]and TGF-β[(634.14±63.58)ng/L vs.(771.62±95.74)ng/L]were significantly lower than those of the control group(P<0.05);the levels of miR-21,Th17,Th17/Treg,IL-6 and IL-17 in cerebrospinal fluid of children with viral encephalitis in se‐vere group were significantly higher than those in mild group(P<0.05),while the levels of Treg,IL-10 and TGF-βwere significantly lower(P<0.05);the levels of miR-21 and Th17/Treg were positively correlated with the levels of IL-6 and IL-17,the level of miR-21 was positively correlated with Th17/Treg(P<0.05),and the levels of miR-21 and Th17/Treg were negatively correlated with the levels of IL-10 and TGF-β(P<0.05).Conclusion The increased expression of miR-21 is closely related to the imbalance of Th17/Treg cell bal‐ance,this process may involve the participation of IL-6,IL-10,IL-17 and TGF-β,and it may be involved in the pathogenesis and aggra‐vation of viral encephalitis.