UVB对系统性红斑狼疮患者外周血单个核细胞自噬相关转录组学的影响

Effects of UVB on Autophagy-related Transcriptomics in Peripheral Blood Mononuclear Cells of Patients with Systemic Lupus Erythematosus

目的检测中波紫外线(UVB)照射对系统性红斑狼疮(SLE)患者的外周血单个核细胞(PBMC)自噬相关mRNA表达的影响并验证。方法分离15例SLE患者和15例正常对照的PBMC,各分为UVB照射组和未照射组,照射UVB后培养24 h,提取细胞总RNA,使用RNA-Seq测序,筛选出各组间自噬相关差异性表达mRNA,并通过qPCR及Western blot对部分基因(ATG7、p62/SQSTM1)的表达情况进行验证。结果RNA-Seq显示SLE患者未照射UVB组(SLE-C组)和正常人未照射UVB组(NC-C组)相比,共有266个基因上调,85个基因下调;SLE患者照射UVB组(SLE-UVB组)和正常人照射UVB组(NC-UVB组)相比,共有318个基因上调,146个基因下调;GO功能富集分析显示差异基因与自噬小体组装、巨自噬的正性调节、巨自噬的调节等有关。对ATG7和p62进行RT-qPCR和Western blot验证,结果显示ATG7和p62在SLE-UVB组的表达水平较SLE-C组和NC-UVB组高,与RNA-Seq的变化趋势一致。结论获得SLE患者PBMC照射UVB前后的自噬相关mRNA表达数据,为进一步阐明UVB对SLE发病的影响提供了理论基础。

Objective To explorer the effect of ultraviolet B(UVB) irradiation on the expression of autophagy-related mRNAs in peripheral blood mononuclear cells(PBMC) of patients with systemic lupus erythematosus(SLE).Methods PBMCs from 15 patients with SLE and 15 normal controls were isolated and divided into UVB-irradiated and non-irradiated groups.After UVB irradiation for 24 hours,total RNA was extracted and sequenced by RNA-Seq.Differentially expressed of mRNAs were screened and part of which(ATG7,p62/SQSTM1) were further detected by qPCR and Western blot.Results RNA-Seq showed that 266 genes were up-regulated and 85 genes were down-regulated in PBMCs from SLE patients unexposed to UVB group(SLE-C group) compared with normal control non-UVB group(NC-C group),318 genes were up-regulated and 146 genes were down-regulated in PBMCs from SLE patients exposed to UVB group(SLE-UVB group) compared with normal control UVB group(NC-UVB group).GO function enrichment analysis showed that differential genes were involved in autophagosome assembly,positive regulation of macroautophagy,and regulation of macroautophagy.RT-qPCR and Western blot of ATG7 and p62 showed that the differentially expression levels of ATG7 and p62 in SLE-UVB group were higher than those in SLE-C group and NC-UVB group.Conclusion The differentially expressed autophagy-related mRNAs in PBMCs of patients with SLE may be a basis for further elucidation of the effect of UVB on the pathogenesis of SLE.