PFKFB3在急性心肌梗死时多形核髓源性抑制细胞炎性活化中的作用  

Effect of PFKFB3 on inflammatory activation of polymorphonuclear myeloid-derived suppressor cell in acute myocardial infarction

作　　者：虞宗颖 吴艳君 刘大东 Yu Zongying;Wu Yanjun;Liu Dadong(Department of Electrocardiograph,Zhenjiang Fourth People's Hospital,Zhenjiang 212001,Jiangsu,China;Department of Cardiology,Zhenjiang Fourth People's Hospital,Zhenjiang 212001,Jiangsu,China;Department of Critical Care Medicine,General Hospital of the Eastern Theater Command of the People's Liberation Army of China,Nanjing 210002,Jiangsu,China)

机构地区：[1]镇江市第四人民医院心电图室,江苏镇江212001 [2]镇江市第四人民医院心内科,江苏镇江212001 [3]中国人民解放军东部战区总医院重症医学科,江苏南京210002

出　　处：《中华危重病急救医学》2024年第1期44-49,共6页Chinese Critical Care Medicine

基　　金：国家自然科学基金(82202389)。

摘　　要：目的探讨6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3(PFKFB3)与急性心肌梗死(AMI)时多形核髓源性抑制细胞(PMN-MDSC)炎性活化的相关性,评价靶向PFKFB3的干预措施在AMI时PMN-MDSC炎性活化中的作用。方法①临床试验部分:采用观察性研究方法,选择镇江市第四人民医院收治的急性冠脉综合征(ACS)患者作为研究对象,根据临床诊断结果分为AMI组和非AMI组。取两组患者外周静脉血,检测PMN-MDSC比例,采用实时定量聚合酶链反应(RT-qPCR)检测单个核细胞PFKFB3的基因表达量。②基础实验部分:将30只6~8周龄雄性C57小鼠按随机数字表法分为正常对照组(n=5)、假手术(Sham)组(n=5)、AMI模型组(n=10)和PFKFB3抑制剂PKF-15干预组(n=10)。采用结扎左冠状动脉前降支(LADCA)的方法建立小鼠AMI模型;Sham组小鼠开胸后不结扎动脉;PKF-15干预组于结扎LADCA的同时,腹腔注射PKF-15(20μg/g)进行干预;正常对照组小鼠不进行任何处理。制模后24 h取小鼠外周静脉血,随后处死小鼠取心肌组织,分别检测小鼠外周血PMN-MDSC比例和心肌组织PMN-MDSC浸润量;苏木素-伊红(HE)染色后,光镜下观察小鼠心肌组织炎症性损伤程度;采用流式细胞仪分选出小鼠PMN-MDSC,并采用RT-qPCR法检测PFKFB3及相关炎症因子基因表达量。结果①临床试验部分:AMI组(n=25)患者外周血PMN-MDSC比例较非AMI组(n=20)明显升高〔(8.53±0.96)%比(1.13±0.39)%,P<0.01〕,外周血单个核细胞PFKFB3基因表达量显著高于非AMI组(2^(-ΔΔCt):1.18±0.19比0.96±0.16,P<0.01);Pearson相关分析显示,AMI患者外周血PMN-MDSC比例与单个核细胞PFKFB3基因表达量呈显著正相关(r=0.608,P=0.001)。②基础实验部分:AMI小鼠外周血及心肌组织PMN-MDSC比例均较正常对照组和Sham组明显增高;PFK-15干预后能够明显降低AMI小鼠外周血及心肌组织PMN-MDSC比例〔外周血:(26.33±5.27)%比(75.12±5.02)%,心肌组织:(20.87±2.97)%比(35.28±4.36)%,均P<0.01〕。光镜Objective To investigate the correlation between 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3(PFKFB3)and the inflammatory activation of polymorphonuclear myeloid-derived suppressor cell(PMN-MDSC)in acute myocardial infarction(AMI),and to evaluate the effect of intervention targeting PFKFB3 on the inflammatory activation of PMN-MDSC during AMI.Methods①Clinical trial section:a observational study was conducted.The patients with acute coronary syndrome(ACS)admitted to Zhenjiang Fourth People's Hospital were enrolled,and they were divided into AMI group and non-AMI group according to clinical diagnosis.The peripheral venous blood of the two groups was collected to detect the proportion of PMN-MDSC,and the expression of PFKFB3 gene in mononuclear cells was detected by real-time quantitative polymerase chain reaction(RT-qPCR).②Basic experiment section:a total of 30 male C57 mice(aged 6-8 weeks)were divided into normal control group(n=5),Sham group(n=5),AMI model group(n=10)and PFKFB3 inhibitor PKF-15 intervention group(n=10)according to random number table method.The AMI model of mice was reproduced by left anterior descending coronary artery(LADCA)ligation,and the mice in the Sham group did not attach the artery after thoracotomy.The PKF-15 intervention group was intraperitoneally injected with PKF-15(20μg/g)at the same time of LADCA ligation.Normal control mice did not receive any treatment.Peripheral venous blood and myocardial tissue of mice were collected 24 hours after modeling.Both the circulating PMN-MDSC ratio and the infiltration of PMN-MDSC in myocardial tissue were detected.After staining with hematoxylin-eosin(HE),the degree of inflammatory damage in mouse myocardial tissue was observed under light microscopy.PMN-MDSC were isolated from mice with flow cytometry,and the gene expressions of PFKFB3 and inflammatory factors were measured by RT-qPCR.Results①Clinical trial section:the circulating PMN-MDSC ratio of patients in the AMI group(n=25)was significantly higher than that in the non-AMI gr

关 键 词：急性心肌梗死 多形核髓源性抑制细胞 炎性活化 PFKFB3 

分 类 号：R542.22[医药卫生—心血管疾病]