激素敏感型及依赖型肾病综合征患儿基因甲基化研究及其生物信息学分析  

Differentially methylated genes in children with steroid-dependent or steroid-sensitive nephrotic syndrome:A bioinformatic analysis

作　　者：张佳仪[1] 芮淑敏 高春林[1] 夏正坤[1] 

机构地区：[1]南方医科大学金陵医院(南京军区南京总医院)儿科,南京医学硕士研究生210002

出　　处：《医学研究生学报》2017年第3期266-270,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81270800)

摘　　要：目的目前关于激素敏感型肾病综合征(SSNS)及激素依赖型肾病综合征(SDNS)患儿的基因甲基化研究较少,文中旨在筛选SSNS和SDNS甲基化差异表达的基因,并进行生物信息学分析,探讨可能的发病机制及寻找潜在的治疗靶标。方法选择2015年1月至12月在南京军区南京总医院儿科住院患者7例,SSNS者(n=3):泼尼松足量[2 mg/(kg·d)或60mg/(m2·d)]治疗,4周内尿蛋白转阴者;SDNS者(n=4):对激素治疗敏感,但是连续2次减药或停药2周内复发者。分别提取2组患儿的外周血DNA,利用基因甲基化芯片技术筛选差异表达的甲基化基因,并对其进行生物信息学对检测结果进行分析。结果甲基化芯片结果发现,与SSNS组比较,SDNS组存在318个差异甲基化基因,其中有193个基因表达为高甲基化,125个为低甲基化;这些异常基因主要位于DNA的开放阅读框和Cp G岛区域。甲基化差异基因主要涉及Rho鸟嘌呤核苷酸交换因子活性、核苷三磷酸酶调节活性、GTP酶调节活性等分子功能。生物学过程方面主要涉及调节前体代谢产物和能量、抗原加工提呈、Rho及Ras蛋白信号传导通路、伪足集合、再生等生物过程。在细胞组成方面,主要涉及MHC蛋白复合物、染色质周纤维及MHC 1类分子的蛋白复合物。通过KEGG信号通路分析发现,差异基因主要参与9个信号通路,其中主要涉及I型糖尿病、淀粉与蔗糖的代谢、同种异体移植物的排斥和自身免疫性甲状腺病等信号通路。结论广泛存在的SDNS患儿基因异常甲基化现象,为激素依赖的原因之一,这为寻找潜在的治疗靶标提供了依据。Objective Few researches have been reported on the gene methylation in children with steroid-sensitive nephrotic syndrome （SSNS） or steroid-dependent nephrotic syndrome （SDNS）. This study aimed to investigate the possible pathogenesis and therapeutic target of SSNS and SDNS by screening differentially methylated genes （DMGs） and bioinformatic analysis using DNA methylation microarray. Methods This study included 3 hospitalized children with SSNS and another 4 with SDNS, all treated with full dose of prednisone （ 2 mg per kilogram of the body weight per day or 60 mg per m^2 per day ） . Negative urine protein was achieved within 4 weeks in the former group, while the latter, though sensitive to hormonal therapy, relapsed within 2 weeks after drug withdrawal or dose reduction. DNA was extracted from the peripheral blood of the patients in both groups for screening DMGs and bioinformatic analysis using DNA methylation microarray. Results Compared with the patients with SSNS, 318 DMGs were found in the SDNS group, among which 193 were hypermethylated and the other 125 hypomethylated. These abnormal genes were mainly located in the open reading frame of DNA and the CpG island region. DMGs were mainly involved in Rho guanyl-nucleotide exchange factor activity, nucleoside- triphosphatase regulator activity, GTPase activator activity, and other molecular functions. The biological processes were chiefly associated with the regulation of the generation of precursor metabolites and energy, antigen processing and presentation, regulation of Rho and Ras protein signal transduction, lamellipodium assembly, regeneration, and other biological processes. The cell composition was mainly related to MHC protein complexes, perichromatin fibrils, and the MHC class Ⅰ protein complex. Analysis of the KEGG signaling pathway showed that DMGs participated in 9 signaling pathways, involving type Ⅰ diabetes, starch and sucrose metabolism, al

关 键 词：原发性肾病综合征 甲基化芯片 激素依赖 

分 类 号：R692[医药卫生—泌尿科学]