摘要
目的探究线粒体DNA(mtDNA)的D-loop区单核苷酸多态性(SNPs)和mtDNA拷贝数与皮肌炎(DM)发病风险的关系及其影响因素。方法收录74例DM患者和92例健康受试者,从外周血液中提取基因组DNA,利用PCR技术将mtDNA的D-loop区目的片段进行扩增,对产物进行测序;应用高灵敏度活性氧(ROS)检测套件,测定血清ROS水平;采用流式免疫荧光微球技术测定细胞因子白细胞介素(IL)-5、IL-13、干扰素(IFN)-γ、IL-2、IL-6、IL-10、肿瘤坏死因子-α(TNF-α)、IL-4表达水平,使用Wilcoxon秩和检验评估细胞因子与DM风险相关SNPs的潜在相关性;进行实时荧光定量聚合酶链反应(qPCR)分析,测量mtDNA的相对拷贝数。结果DM患者组存在2个与其发病风险有关的多态性变异位点(16304T/C、16519T/C),线粒体D-loop区的等位基因16304C(χ^(2)=4.937,P=0.026)和16519C(χ^(2)=4.405,P=0.036)与DM患者发病风险有关;DM风险相关等位基因16304C与IL-4低表达相关(P=0.016)。DM患者中的mtDNA拷贝数高于对照组(P<0.001)。结论线粒体D-loop区SNPs可能是DM风险的潜在生物标志物,SNPs可能通过影响细胞因子参与DM的发生。DM的mtDNA拷贝数呈现高表达,mtDNA拷贝数的增加可能会导致线粒体功能障碍,从而引发DM的发病。
Objective To explore the relationship between single nucleotide polymorphisms(SNPs)in D-loop region of mitochondrial DNA(mtDNA)and mtDNA copy number and the risk of dermatomyositis(DM),and its influencing factors.Methods 74 patients with DM and 92 healthy controls were included in the study.Genomic DNA was extracted from peripheral blood and the target fragment of mtDNA D-loop region was amplified by PCR technique,and the products were subsequently sequenced.Serum levels of ROS were assessed using a high-sensitivity reactive oxygen species detection kit.The expression levels of cytokines,interleukin(IL)-5,IL-13,interferon-γ(IFN-γ),IL-2,IL-6,IL-10,tumor necrosis factor-α(TNF-α)and IL-4 were measured using Flow Fluorescence Immunmicrobeads Assay.Wilcoxon rank-sum test was used to assess the potential correlation between cytokines and SNPs associated with DM risk.The relative copy number of mtDNA was measured using quantitative real-time polymerase chain reaction(qPCR)analysis.Results Two SNPs(16304T/C,16519T/C)were found to be associated with the risk of developing DM,and alleles 16304C(χ^(2)=4.937,P=0.026)and 16519C(χ^(2)=4.405,P=0.036)in the mitochondrial D-loop region were confirmed to be associated with DM development risk.The DM risk-associated allele 16304C was significantly associated with lower IL-4 expression(P=0.016).The mtDNA copy number was significantly higher in DM patients than in controls(P<0.001).Conclusion Mitochondrial D-loop SNPs can be potential biomarkers for DM risk,and SNPs may be involved in DM by influencing cytokines.DM shows high expression of mtDNA copy number,and the increase in mtDNA copy number may lead to mitochondrial dysfunction,which triggers the pathogenesis of DM.
作者
檀紫瑞
张晶晶
贾园园
彭晨星
赵宇飞
Tan Zirui;Zhang Jingjing;Jia Yuanyuan;Peng Chenxing;Zhao Yufei(Dept of Thoracic Surgery,The Fourth Hospital of Hebei Medical University,Shijiazhuang 050011;Dept of Immunology and Rheumatology,The Fourth Hospital of Hebei Medical University,Shijiazhuang 050011;Dept of Immunology and Rheumatology,The Second Hospital of Hebei Medical University,Shijiazhuang 050000)
出处
《安徽医科大学学报》
北大核心
2025年第1期130-135,共6页
Acta Universitatis Medicinalis Anhui
基金
河北省医学科学研究课题计划(编号:20221248)。
作者简介
檀紫瑞,男,主治医师;通信作者:赵宇飞,男,主治医师,E-mail:zhaoyufei724@aliyun.com。
