免疫相关基因在绝经后骨质疏松症患者外周血白细胞中的表达

文题释义： RNA-seq技术：利用高通量测序技术进行转录组测序分析，检测mRNA、miRNA、circRNA及lncRNA等所有转录本的表达水平，从而探究特定组织或细胞中基因的结构和功能，探寻特定疾病发生、发展的分子机制和信号通路。 骨免疫学：2000年首次提出，认为骨骼是一种免疫器官，在骨髓中形成的多种免疫细胞可与骨骼系统的细胞相互作用，通过经典的RANKL/RANL/OPG系统等信号通路共同调控骨代谢过程。最近提出的“免疫疏松”，进一步揭示骨免疫学在骨质疏松症中的调节作用。   摘要 背景：目前认为骨骼是一种免疫器官，在骨髓中形成的多种免疫细胞可与骨骼系统的细胞相互作用，共同调控骨代谢。研究绝经后骨质疏松症的发病机制，寻找其治疗的分子靶标和信号通路，有助于其预防和治疗。 目的：利用RNA-Seq技术，探讨绝经后骨质疏松症患者外周血白细胞中与免疫相关的基因表达谱的变化。 方法：选取因发生骨折住院治疗的绝经后0-20年的女性40人，首先采用Lunar Prodigy双能X射线骨密度仪扫描后，分为骨量正常组(T>-1)和骨质疏松组(T<-2.5)。全转录组测序(RNA-seq)分别抽取2组各5人的外周血样本，分离白细胞后提取总RNA，筛选2组的表达差异基因并进行GO富集分析，筛选免疫相关表达基因并进行KEGG信号通路分析，收集2组各20人的外周血样本采用Real-time PCR验证KEGG PATHWAY生物信息学分析的结果。研究方案的实施符合上海市浦东新区公利医院的相关伦理要求(20170301)。 结果与结论：①骨质疏松组与骨量正常组相比，表达差异在2倍以上且具有统计学意义的基因为187个(其中表达上调131个，表达下调56个)，其中与免疫相关的表达差异基因为29个(其中表达上调25个，表达下调4个)；②KIR3DL1、KIR3DL2、KIR2DL4、KLRD1及HSPA6在2组中的表达差异有显著性意义(P < 0.05)；③结果说明，KEGG信号通路分析显示上述5个基因在自然杀伤细胞介导的细胞毒作用中发挥重要作用。KIR3DL1、KIR3DL2、KIR2DL4、KLRD1及HSPA6参与的自然杀伤细胞介导的细胞毒作用可能与绝经后骨质疏松症的发生密切相关。ORCID: 0000-0003-4879-0905(陈天宁) 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Expression of immune-related genes in peripheral blood leukocytes of postmenopausal osteoporosis patients

BACKGROUND:Bones are currently considered as an immune organ.A variety of immune cells that originate from the bone marrow can interact with the cells of the skeletal system to jointly regulate bone metabolism.Explorations on the pathogenesis of postmenopausal osteoporosis as well as treatment-related molecular targets and signal pathways can help prevention and treatment of the disease.OBJECTIVE:To investigate the expression profiles of immune-related genes in peripheral blood leukocytes of postmenopausal osteoporosis patients using RNA-Seq technology.METHODS:Forty female patients who had experienced menopause for 0 to 20 years and were hospitalized due to fractures were enrolled.They were divided into normal bone mass group(T>-1)and osteoporosis group(T<-2.5)by scanning with Lunar Prodigy dual-energy X-ray bone densitometer.Then,the total RNA of leukocytes in the peripheral blood samples from five patients of each group was extracted.Differentially expressed genes between two groups were detected by RNA-Seq technology followed by GO enrichment analysis.The immune-related genes were screened and analyzed by KEGG signal pathway.Peripheral blood samples from 20 patients of each group were collected and the results of KEGG PATHWAY bioinformatics analysis were verified by real-time PCR.The implementation of the study plan complied with the relevant ethical requirements of the Pudong New Area Gongli Hospital,Shanghai(approval No.20170301).RESULTS AND CONCLUSION:Compared with the normal bone mass group,187 genes were significantly changed in the osteoporosis group(fold change>2),and 131 genes were up-regulated and 56 genes were down-regulated.We identified in total 29 differentially expressed immune-related genes including 25 up-regulated and 4 down-regulated ones.There was significant difference in expression between the osteoporosis and normal bone mass groups for genes,including KIR3DL1,KIR3DL2,KIR2DL4,KLRD1 and HSPA6(P<0.05).These differentially expressed genes are potentially important for the natural killer cell-mediated cytotoxicity by the KEGG pathway analysis.KIR3DL1,KIR3DL2,KIR2DL4,KLRD1 and HSPA6 may be closely related to the natural killer cell-mediated cytotoxicity during the occurrence of postmenopausal osteoporosis.