上海交通大学学报（医学版）

上海交通大学学报（医学版） >

2021 , Vol. 41 >Issue 12: 1603 - 1611

DOI: https://doi.org/10.3969/j.issn.1674-8115.2021.12.009

论著 · 基础研究

顺铂诱导的急性肾损伤中肾脏组织m6A甲基化水平的变化

  • 沈剑箫 ,
  • 王万鹏 ,
  • 邵兴华 ,
  • 吴晶魁 ,
  • 李舒 ,
  • 车霞静 ,
  • 倪兆慧
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  • 1.上海交通大学医学院附属仁济医院肾脏科，上海 200127
    2.江苏省涟水县人民医院肾脏科，涟水 223400
沈剑箫（1985—），男，主治医师，博士；电子信箱：shenjianxiao@aliyun.com。
倪兆慧，电子信箱：profnizh@126.com。

收稿日期: 2021-08-12

  网络出版日期: 2021-01-28

基金资助

国家自然科学基金(81700586);上海市核酸化学与纳米医学重点实验室资助计划(2020ZYB009);上海申康医院发展中心临床三年行动计划(SHDC2020CR3029B);上海市卫生健康委员会科研基金(ZHYY-ZXYJHZX-202014);上海市科学技术委员会基金(13401906100)

收起

Changes of m6A methylation in renal tissue during cisplatin-induced acute injury

  • Jian-xiao SHEN ,
  • Wan-peng WANG ,
  • Xing-hua SHAO ,
  • Jing-kui WU ,
  • Shu LI ,
  • Xia-jing CHE ,
  • Zhao-hui NI
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  • 1.Department of Nephrology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
    2.Department of Nephrology, Lianshui People's Hospital, Jiangsu Province, Lianshui 223400, China
NI Zhao-hui, E-mail: profnizh@126.com.

Received date: 2021-08-12

  Online published: 2021-01-28

Supported by

National Natural Science Foundation of China(81700586);Key Shanghai Laboratory of Nucleic Acid Chemistry and Nanomedicine(2020ZYB009);Clinical Research Plan of Shenkang Hospital Development Center(SHDC2020CR3029B);Science and Research Fund of Shanghai Municipal Health Commission(ZHYY-ZXYJHZX-202014);Fund from Science and Technology Commission of Shanghai Municipality(13401906100)

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摘要

目的·探讨N6-甲基腺嘌呤（N6-methyladenosine，m6A）甲基化修饰在顺铂诱导的小鼠急性肾损伤进程中的作用。方法·选择4只C57bL/6小鼠，尾静脉注射顺铂（20 mg/kg）诱导急性肾损伤（损伤组）；另取4只C57bL/6小鼠，尾静脉注射等量生理盐水（对照组）。检测2组小鼠血清肌酐及血尿素氮水平变化，观察小鼠肾脏组织切片中病理损伤情况，评估模型是否成功。进一步运用甲基化RNA免疫共沉淀技术（methylated RNA immunoprecipitation，MeRIP）与RNA测序技术分别检测2组小鼠肾脏组织中m6A甲基化水平与RNA表达变化。运用基因本体论及京都基因和基因组数据库进行结果可视化和综合研究，并将RNA测序技术所得转录组数据与MeRIP技术检测所得表观遗传数据联合分析，寻找参与顺铂诱导急性肾损伤病理变化过程的候选基因。结果·顺铂可诱导小鼠血清肌酐与血尿素氮水平显著升高。光学显微镜观察肾组织发现广泛的肾小管空泡变性，上皮细胞剥脱，肾小管坏死，提示造模成功。MeRIP检测发现损伤组与对照组小鼠肾脏中共有2 227个基因含有2 981个差异化表达的m6A甲基化位点（表达变化倍数≥2且P<0.05），这些基因主要富集于代谢及细胞死亡通路。表达差异化m6A甲基化位点的基因与RNA差异化表达基因的联合分析发现1 002个表达趋势相同的基因，如纤维蛋白原α链、溶质载体12家族成员1和甲肝病毒细胞受体1等。结论·顺铂可诱导肾脏组织中基因mRNA上m6A甲基化位点的甲基化水平变化，促进急性肾损伤进程。

关键词： N6-甲基腺嘌呤甲基化; 顺铂诱导急性肾损伤; 纤维蛋白原α链; 溶质载体12家族成员1; 甲肝病毒细胞受体1

本文引用格式

沈剑箫 , 王万鹏 , 邵兴华 , 吴晶魁 , 李舒 , 车霞静 , 倪兆慧 . 顺铂诱导的急性肾损伤中肾脏组织m6A甲基化水平的变化[J]. 上海交通大学学报（医学版）, 2021 , 41(12) : 1603 -1611 . DOI: 10.3969/j.issn.1674-8115.2021.12.009

Abstract

Objective

·To investigate the role of N6-methyladenosine (m6A) methylation modification in the process of cisplatin-induced acute injury in mice.

Methods

·Four C57BL/6 mice were injected with cisplatin (20 mg/kg) through tail vein (the injury group); Another 4 C57BL/6 mice were injected with the same amount of saline (the control group). The changes of serum creatinine and urea nitrogen levels in the mice and the pathological injury in the renal tissue sections of the mice were evaluated to judge the success of the model. Methylated RNA immunoprecipitation (MeRIP) and RNA sequencing were used to detect the changes of m6A methylation and RNA expression in the kidney tissue of the two groups of mice. Gene ontology and Kyoto Encyclopedia of genes and genomes were used for visualization and comprehensive research. Transcriptome data and epigenetic data were combined to find candidate genes for pathological changes of cisplatin-induced acute injury.

Results

·Cisplatin could induce significant increase in the levels of serum creatinine and urea nitrogen compared with those in the baseline. Light microscope showed extensive tubular vacuolar degeneration, epithelial cell exfoliation and tubular necrosis, suggesting the success of modeling. MeRIP detection showed that a total of 2 277 genes contained 2 981 differentially expressed m6A methylation sites (expression multiple ≥2 and P<0.05) in the kidneys of mice in the injury group and the control group. These genes were mainly concentrated in the metabolic and cell death pathways. The joint analysis of genes expressing differential m6A methylation sites and RNA differential expression genes found 1 002 genes with the same expression trend, such as fibrinogen α chain, solute carrier family 12 member 1 and hepatitis A virus cellular receptor 1.

Conclusion

·Cisplatin can induce the change of methylation level of m6A methylation site on mRNA in renal tissue, and promote the process of acute renal injury.

Key words： N6-methyladenosine (m6A) methylome; cisplatin-induced acute kidney injury (CI-AKI); fibrinogen α chain (FGA); solute carrier family 12 member 1 (Slc12a1); hepatitis A virus cellular receptor 1 (Harvc1)

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