早幼粒细胞白血病锌指在哮喘患者外周血中的表达及意义

The expression and clinical signifi cance of promyelocytic leukemia zinc fi nger in human peripheral blood with asthma

目的检测早幼粒细胞白血病锌指结构(promyelocytic leukemia zinc fi nger,PLZF)在哮喘(asthma)患者外周血中的表达水平及临床意义。方法选取上海市第五人民医院2021年5月至2021年10月诊治的稳定期Asthma患者外周血标本为研究对象(n=40),以及健康体检者为对照组(Control组,n=40),采用酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)检测血清中细胞因子的含量,实时荧光定量PCR(quantitative real-time PCR,qPCR)检测血浆中PLZF的mRNA表达。分离外周血的单个核淋巴细胞(peripheral blood mononuclear cell,PBMC),并使用流式细胞术检测PBMC中PLZF+细胞的水平和分布。采用SPSS 26.0及Graphpad Prism 7.0软件进行独立样本t检验、Mann-Whitney U检验、χ2检验、ROC曲线及Logistic回归等统计学分析。以P<0.05为差异有统计学意义。结果与Control组相比,在Asthma组中细胞因子干扰素-γ(interferonγ,IFN-γ)、白细胞介素-2(interleukin-2,IL-2)、IL-4、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和IL-17表达水平增加,差异有统计学意义(P<0.05),而IL-10两组相比差异无统计学意义。Asthma组PBMC中PLZF的mRNA表达明显增高(P<0.05),PLZF+细胞明显增加(3.40±2.52%,Control组:1.23±0.78%,P<0.05),其中以CD8+PLZF+及Vβ11+PLZF+T细胞为主,提示PLZF主要在CD8+、恒定自然杀伤T细胞(invariant natural killer T cell,iNKT)细胞亚群中表达增加。Logistic回归分析及ROC曲线结果显示,PBMC中PLZF+细胞增高显著增加了哮喘发生的风险(OR=5.76>1,AUC=0.87,P<0.05)。结论哮喘患者外周血中PLZF增加,可能在哮喘疾病发展中起到重要作用,结果尚需更大样本实验数据进一步证实。

Objective To investigate the expression of promyelocytic leukemia zinc finger(PLZF)in human peripheral blood with asthma and its clinical signifi cance.Methods Forty patients with stable asthma from May 2021 to October 2021 in the Department of Respiratory Medicine of the Shanghai Fifth People's Hospital were enrolled,and forty healthy controls were recruited in the study.The levels of cytokines in serum were measured by enzyme-linked immunosorbent assay(ELISA).Quantitative real-time PCR(qPCR)was used to detect the expression of PLZF mRNA in plasma.The level and distribution of PLZF+cells in PBMCs were detected by fl ow cytometry after isolating peripheral blood mononuclear cells(PBMCs).Independent sample t test,Mann-Whitney U test,χ2 test,ROC curve and Logistic regression were used to analyze the results with SPSS 26.0 and Graphpad Prism 7.0.A P<0.05 was considered statistically significant.Results The levels of cytokines IFN-γ,IL-2,IL-4,TNF-αand IL-17 in human peripheral blood from the asthma group were obviously higher than those in the control group(P<0.05),whereas there was no signifi cant difference in the level of cytokine IL-10 between the two groups.The level of PLZF mRNA in PBMCs from the asthma group was signifi cantly upregulated compared to that in the control group[(3.40±2.52%)vs.(1.23±0.78%),P<0.05].CD8+PLZF+and Vβ11+PLZF+T cells in the asthma group were signifi cantly outnumbered than those in the control group(P<0.05).Logistic regression and ROC curve analysis showed that PLZF expression in PBMC was a risk factor for the development of asthma(OR>1,AUC=0.87,P<0.05).Conclusions The high expression of PLZF in peripheral blood may play an important role in the development of asthma,which needs to be further confi rmed by large sample studies.