中国组织工程研究

中国组织工程研究 ›› 2021, Vol. 25 ›› Issue (5): 741-745.doi: 10.3969/j.issn.2095-4344.3009

• 组织构建临床实践 clinical practice in tissue construction • 上一篇    下一篇

肾移植患者血清中长链非编码RNA PGM5-AS1表达及调控人肾小球内皮细胞的作用

蒋  欣，乔良伟，孙  东，李  明，房  军，曲青山   

  1. 郑州人民医院器官移植科，河南省郑州市   450003
  • 收稿日期:2020-02-10 修回日期:2020-02-17 接受日期:2020-04-11 出版日期:2021-02-18 发布日期:2020-11-28
  • 通讯作者: 曲青山，主任医师，教授，郑州人民医院器官移植科，河南省郑州市 450003
  • 作者简介:蒋欣，男，1980年生，汉族，河南省永城市人，2007年郑州大学毕业，硕士，副主任医师，主要从事肾脏移植的基础与临床研究。
  • 基金资助:
    河南省医学科技攻关计划联合共建项目(2018020823)

Expression of long chain non-coding RNA PGM5-AS1 in serum of renal transplant patients and its regulation of human glomerular endothelial cells

Jiang Xin, Qiao Liangwei, Sun Dong, Li Ming, Fang Jun, Qu Qingshan   

  1. Department of Organ Transplantation, Zhengzhou People’s Hospital, Zhengzhou 450003, Henan Province, China
  • Received:2020-02-10 Revised:2020-02-17 Accepted:2020-04-11 Online:2021-02-18 Published:2020-11-28
  • Contact: Qu Qingshan, Chief physician, Professor, Department of Organ Transplantation, Zhengzhou People’s Hospital, Zhengzhou 450003, Henan Province, China
  • About author:Jiang Xin, Master, Associate chief physician, Department of Organ Transplantation, Zhengzhou People’s Hospital, Zhengzhou 450003, Henan Province, China
  • Supported by:
    the Medical Science Tackle Key Plan of Henan Province, No. 2018020823

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摘要：

文题释义：
巨噬细胞趋化性：巨噬细胞是一种来源于骨髓中的单核细胞，由前体细胞分化而来。巨噬细胞参与调解血细胞发育、胚胎期器官发育、血管生成、细胞凋亡和免疫排斥等；巨噬细胞沿着定向趋化因子浓度增加的信号向趋化因子源处的迁徙。
急性排斥反应：是由于受体识别供体抗原引起，主要是T淋巴细胞介导的免疫反应，效应细胞多为细胞毒性T细胞和记忆性T细胞，它们可能通过直接提呈方式被激活。急性排斥反应属于迟发性超敏反应，分为细胞性排斥反应和血管性排斥反应；应用抗排斥治疗多可逆转。

背景：lnc RNA PGM5-AS1在肾移植急性免疫排斥患者外周血清中表达降低，其表达对肾小球内皮细胞存活和巨噬细胞趋化性的影响还有待研究。
目的：探究lnc RNA PGM5-AS1在肾移植急性排斥反应患者和非急性排斥反应患者血清中的表达及其对肾小球内皮细胞(HRGEC)增殖、细胞周期、细胞凋亡及巨噬细胞趋化性的影响。
方法：①肾移植患者46例，根据术后1个月内发生急性排斥反应与否分为急性排斥反应组(17例)和非急性排斥反应组(29例)，术前1 d及术后第1，2，3，4周各抽取外周血，qRT-PCR检测患者血清中PGM5-AS1表达；②将si-control和沉默PGM5-AS1(si-PGM5-AS1)转染至人肾小球内皮细胞(HRGEC细胞)，以空白组为对照；qPCR检测转染后细胞中PGM5-AS1表达；MTT检测细胞增殖，流式细胞术检测细胞凋亡和细胞周期，ELISA检测细胞炎症因子分泌，Transwell检测巨噬细胞趋化性。研究方案经郑州人民医院伦理学委员会批准，所有患者签署知情同意书。
结果与结论：①与非急性排斥反应患者相比，肾移植急性排斥反应患者移植后1，2，3，4周血清中PGM5-AS1表达降低(P < 0.05)；②沉默PGM5-AS1后HRGEC细胞中PGM5-AS1表达低于抑制si-control组和空白组(F=379.658，P < 0.05)；③MTT结果显示，沉默PGM5-AS1可显著抑制HRGEC细胞增殖(P < 0.05)；④流式细胞术检测显示，沉默PGM5-AS1诱导HRGEC细胞阻滞在G0/G1期和增加细胞凋亡(P < 0.05)；⑤ELISA结果显示，沉默PGM5-AS1抑制白细胞介素13表达，增加白细胞介素6、γ干扰素和肿瘤坏死因子α表达(P < 0.05)；⑥Transwell检测显示，沉默PGM5-AS1的HRGEC细胞增加巨噬细胞的趋化性(P < 0.05)；⑦结果说明，PGM5-AS1在肾移植急性排斥反应患者血清中低表达，抑制PGM5-AS1能够促进HRGEC细胞损伤，可作为早期急性排斥反应的外周血诊断标志物，并可能成为治疗急性排斥反应的分子靶点。
https://orcid.org/0000-0002-7455-3690 (蒋欣) 

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 肾, 肾移植, 急性排斥反应, 长链非编码RNA, 肾小球内皮细胞, 细胞周期, 靶点, 标志物

Abstract: BACKGROUND: The expression of long chain non-coding RNA PGM5-AS1 (lnc RNA PGM5-AS1) is reduced in peripheral blood of patients with acute immune rejection after renal transplantation. The effect of its expression on human glomerular endothelial cell (HRGEC) survival and macrophage chemotaxis remains to be studied.
OBJECTIVE: To investigate the expression of lnc RNA PGM5-AS1 in serum of patients with acute rejection of renal transplantation and non-acute rejection patients and its effect on proliferation, cell cycle, apoptosis and macrophages chemotropism of HRGECs.
METHODS: Forty-six patients with renal transplantation were divided into acute rejection group (n=17) and non-acute rejection group (n=29) according to whether or not acute rejection occurred within 1 month after operation. Peripheral blood sample was drawn from each patient at 1 day before operation, 1, 2, 3, and 4 weeks after operation. qRT-PCR was used to detect the expression of PGM5-AS1 in serum of patients with or without acute rejection. si-control and si-PGM5-AS1 were transfected into glomerular endothelial cells, and the expression of PGM5-AS1 in the transfected cells was detected by qPCR. MTT was used to detect cell proliferation, flow cytometry was applied to detect apoptosis and cell cycle, ELISA was adopted to detect cellular inflammatory factor secretion, and Transwell was used to detect chemotaxis of macrophages. Approval for this study was obtained from the Ethics Committee of Zhengzhou People’s Hospital, and all patients signed informed consent prior to the participation in the trial.
RESULTS AND CONCLUSION: Compared with non-acute rejection patients, patients with acute rejection to renal transplantation had significantly lower PGM5-AS1 expression in serum at 1, 2, 3, and 4 weeks after transplantation (P < 0.05). After PGM5-AS1 silencing, the expression of GM5-AS1 in HRGEC cells was significantly lower than that in the si-control group and normal control group (F=379.658, P < 0.05). MTT results showed that PGM5-AS1 silencing significantly inhibited HRGEC cell proliferation (P < 0.05). Flow cytometry results showed that PGM5-AS1 silencing induced HRGEC cell arrest in G0/G1 phase and increased apoptosis (P < 0.05). ELISA results showed that PGM5-AS1 silencing inhibited interleukin-13 expression and increased interleukin-6, interferon-γ and tumor necrosis factor-α expression (P < 0.05). Transwell results indicated that HRGEC cells silenced by PGM5-AS1 significantly increased the chemotaxis of macrophages (P < 0.05). All these findings indicate that PGM5-AS1 is lowly expressed in serum of patients with acute rejection of renal transplantation, and inhibition of PGM5-AS1 can promote HRGEC cell damage, which can be used as a peripheral blood diagnostic marker for early acute rejection, and may be a molecular target for the treatment of acute rejection.

Key words: kidney, renal transplantation, acute rejection, long non-coding RNA, glomerular endothelial cells, cell cycle, target, marker

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