摘要
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。
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-γand tumor necrosis factor-αexpression(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.
作者
蒋欣
乔良伟
孙东
李明
房军
曲青山
Jiang Xin;Qiao Liangwei;Sun Dong;Li Ming;Fang Jun;Qu Qingshan(Department of Organ Transplantation,Zhengzhou People’s Hospital,Zhengzhou 450003,Henan Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2021年第5期741-745,共5页
Chinese Journal of Tissue Engineering Research
基金
河南省医学科技攻关计划联合共建项目(2018020823),项目负责人:曲青山。
