目的:探讨湿性年龄相关性黄斑变性(ARMD)出血患者血清长链非编码RNA母系表达基因3(lncRNA MEG3)和微小RNA-138(miR-138)表达水平及其与患者预后的关系。方法:前瞻性研究。选取2018-01/2021-06收治的湿性ARMD出血患者90例为观察组,选择...目的:探讨湿性年龄相关性黄斑变性(ARMD)出血患者血清长链非编码RNA母系表达基因3(lncRNA MEG3)和微小RNA-138(miR-138)表达水平及其与患者预后的关系。方法:前瞻性研究。选取2018-01/2021-06收治的湿性ARMD出血患者90例为观察组,选择同期在我院进行健康体检人员78名作为对照组。采用实时荧光定量PCR法(qRT-PCR)检测所有受试者血清lncRNA MEG3、miR-138表达水平。观察组患者注射雷珠单抗进行治疗,每月1次,共3次。治疗后随访3mo,根据预后情况分为预后良好组和预后不良组,比较两组患者lncRNA MEG3、miR-138水平。Pearson相关性分析lncRNA MEG3与miR-138的相关关系。采用受试者工作特征曲线(ROC)分析湿性ARMD出血患者预后不良的判断价值。多因素Logistic回归分析湿性ARMD出血患者预后不良的影响因素。结果:与对照组相比,观察组患者血清lncRNA MEG3水平下降(1.13±0.37 vs 0.71±0.21),miR-138水平上升(1.05±0.29 vs 2.23±0.54)(均P<0.05)。湿性ARMD出血患者预后良好组患者血清lncRNA MEG3水平明显高于预后不良组(0.81±0.24 vs 0.49±0.14),而miR-138水平明显低于预后不良组(1.92±0.49 vs 2.87±0.63)(均P<0.05)。Pearson相关性分析结果显示,lncRNA MEG3与miR-138呈负相关(r=-0.381,P<0.05)。ROC曲线结果显示,血清lncRNA MEG3、miR-138表达水平对湿性ARMD出血患者发生预后不良AUC分别为0.859、0.828,截断值分别为0.635、2.455,敏感度分别为89.70%、75.90%,特异性分别为72.10%、82.00%。多因素Logistic回归分析显示lncRNA MEG3是湿性ARMD出血患者预后不良的保护因素,miR-138是危险因素(均P<0.05)。结论:血清lncRNA MEG3、miR-138在湿性ARMD出血患者中表达异常,且对湿性ARMD出血患者的预后不良具有一定的评估价值。展开更多
Objective:This research was performed to explore the effect of macrophage migration inhibitory factor(MIF)on the apoptosis of bone marrow mesenchymal stem cells(BMSCs)in ischemia and hypoxia environments.Methods:The c...Objective:This research was performed to explore the effect of macrophage migration inhibitory factor(MIF)on the apoptosis of bone marrow mesenchymal stem cells(BMSCs)in ischemia and hypoxia environments.Methods:The cell viability of BMSCs incubated under hypoxia/ischemia(H/I)conditions with or without pretreatment with MIF or triglycidyl isocyanurate(TGIC)was detected using cell counting kit-8(CCK-8)analysis.Plasmids containing long noncoding RNA(lncRNA)maternally expressed gene 3(MEG3)orβ-catenin small interfering RNA(siRNA)were used to overexpress or downregulate the corresponding gene,and the p53 signaling pathway was activated by pretreatment with TGIC.The influences of MIF,overexpression of lncRNA MEG3,activation of the p53 signaling pathway,and silencing ofβ-catenin on H/I-induced apoptosis of BMSCs were revealed by western blotting,flow cytometry,and terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL)staining.Results:From the results of CCK-8 assay,western blotting,and flow cytometry,pretreatment with MIF significantly decreased the H/I-induced apoptosis of BMSCs.This effect was inhibited when lncRNA MEG3 was overexpressed by plasmids containing MEG3.The p53 signaling pathway was activated by TGIC,andβ-catenin was silenced by siRNA.From western blot results,the expression levels ofβ-catenin in the nucleus and phosphorylated p53(p-p53)were downregulated and upregulated,respectively,when the lncRNA MEG3 was overexpressed.Through flow cytometry,MIF was also shown to significantly alleviate the increased reactive oxygen species(ROS)level of BMSCs caused by H/I.Conclusions:In summary,we conclude that MIF protected BMSCs from H/I-induced apoptosis by downregulating the lncRNA MEG3/p53 signaling pathway,activating the Wnt/β-catenin signaling pathway,and decreasing ROS levels.展开更多
文摘目的:探讨湿性年龄相关性黄斑变性(ARMD)出血患者血清长链非编码RNA母系表达基因3(lncRNA MEG3)和微小RNA-138(miR-138)表达水平及其与患者预后的关系。方法:前瞻性研究。选取2018-01/2021-06收治的湿性ARMD出血患者90例为观察组,选择同期在我院进行健康体检人员78名作为对照组。采用实时荧光定量PCR法(qRT-PCR)检测所有受试者血清lncRNA MEG3、miR-138表达水平。观察组患者注射雷珠单抗进行治疗,每月1次,共3次。治疗后随访3mo,根据预后情况分为预后良好组和预后不良组,比较两组患者lncRNA MEG3、miR-138水平。Pearson相关性分析lncRNA MEG3与miR-138的相关关系。采用受试者工作特征曲线(ROC)分析湿性ARMD出血患者预后不良的判断价值。多因素Logistic回归分析湿性ARMD出血患者预后不良的影响因素。结果:与对照组相比,观察组患者血清lncRNA MEG3水平下降(1.13±0.37 vs 0.71±0.21),miR-138水平上升(1.05±0.29 vs 2.23±0.54)(均P<0.05)。湿性ARMD出血患者预后良好组患者血清lncRNA MEG3水平明显高于预后不良组(0.81±0.24 vs 0.49±0.14),而miR-138水平明显低于预后不良组(1.92±0.49 vs 2.87±0.63)(均P<0.05)。Pearson相关性分析结果显示,lncRNA MEG3与miR-138呈负相关(r=-0.381,P<0.05)。ROC曲线结果显示,血清lncRNA MEG3、miR-138表达水平对湿性ARMD出血患者发生预后不良AUC分别为0.859、0.828,截断值分别为0.635、2.455,敏感度分别为89.70%、75.90%,特异性分别为72.10%、82.00%。多因素Logistic回归分析显示lncRNA MEG3是湿性ARMD出血患者预后不良的保护因素,miR-138是危险因素(均P<0.05)。结论:血清lncRNA MEG3、miR-138在湿性ARMD出血患者中表达异常,且对湿性ARMD出血患者的预后不良具有一定的评估价值。
基金supported by the National Natural Science Foundation of China(No.81702132)the Zhejiang Provincial Natural Science Foundation of China(No.LY21H060007)+1 种基金the Projects of Medical and Health Technology Program in Zhejiang Province(No.2021KY206)the Wenzhou Public Welfare Scienceand Technology Research Project(Nos.Y20190267 and Y20210436),China.
文摘Objective:This research was performed to explore the effect of macrophage migration inhibitory factor(MIF)on the apoptosis of bone marrow mesenchymal stem cells(BMSCs)in ischemia and hypoxia environments.Methods:The cell viability of BMSCs incubated under hypoxia/ischemia(H/I)conditions with or without pretreatment with MIF or triglycidyl isocyanurate(TGIC)was detected using cell counting kit-8(CCK-8)analysis.Plasmids containing long noncoding RNA(lncRNA)maternally expressed gene 3(MEG3)orβ-catenin small interfering RNA(siRNA)were used to overexpress or downregulate the corresponding gene,and the p53 signaling pathway was activated by pretreatment with TGIC.The influences of MIF,overexpression of lncRNA MEG3,activation of the p53 signaling pathway,and silencing ofβ-catenin on H/I-induced apoptosis of BMSCs were revealed by western blotting,flow cytometry,and terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL)staining.Results:From the results of CCK-8 assay,western blotting,and flow cytometry,pretreatment with MIF significantly decreased the H/I-induced apoptosis of BMSCs.This effect was inhibited when lncRNA MEG3 was overexpressed by plasmids containing MEG3.The p53 signaling pathway was activated by TGIC,andβ-catenin was silenced by siRNA.From western blot results,the expression levels ofβ-catenin in the nucleus and phosphorylated p53(p-p53)were downregulated and upregulated,respectively,when the lncRNA MEG3 was overexpressed.Through flow cytometry,MIF was also shown to significantly alleviate the increased reactive oxygen species(ROS)level of BMSCs caused by H/I.Conclusions:In summary,we conclude that MIF protected BMSCs from H/I-induced apoptosis by downregulating the lncRNA MEG3/p53 signaling pathway,activating the Wnt/β-catenin signaling pathway,and decreasing ROS levels.
