程序性死亡受体1及其配体信号通路对心肌缺血再灌注损伤的影响机制  

Effect of programmed cell death-1/programmed death ligand-1 signaling pathway on myocardial ischemia-reperfusion injury

作　　者：马小兰 赵济尧 郑艳 尤力吐孜·艾尼瓦尔 王俊杰 徐维昉[3] 李杰[3] Ma Xiaolan;Zhao Jiyao;Zheng Yan;Yulituzi·Einivar;Wang Junjie;Xu Weifang;Li Jie(Nursing College of Xinjiang Medical University,Xinjiang 830054,China;First Clinical Medical College of Xinjiang Medical University,Xinjiang 830054,China;Department of Anesthesiology,the First Affiliated Hospital of Xinjiang Medical University,Xinjiang 830054,China)

机构地区：[1]新疆医科大学护理学院,乌鲁木齐830054 [2]新疆医科大学第一临床医学院,乌鲁木齐830054 [3]新疆医科大学第一附属医院麻醉科,乌鲁木齐830054

出　　处：《中华实验外科杂志》2024年第8期1699-1702,共4页Chinese Journal of Experimental Surgery

基　　金：新疆维吾尔自治区自然科学基金(2021D01C308)。

摘　　要：目的探讨程序性死亡受体1及其配体信号通路对心肌缺血再灌注损伤影响以及机制。方法采用随机数字表法将20只8~10周龄C57BL小鼠,分为假手术组(Sham组)、缺血再灌注组(I/RI组),I/RI+CPG-ODN组和程序性死亡因子配体1(PD-L1)单抗处理组(I/RI+PD-L1组),每组5只,送单细胞测序,检测白细胞介素(IL)-2、IL-6、γ-干扰素(IFN-γ),蛋白印迹法检测组织中PD-L1、细胞凋亡蛋白B细胞淋巴瘤白血病-2(bcl-2)及bcl-2相关X蛋白(bax)的蛋白表达;将H9C2心肌细胞随机分为正常对照组(Control组)、缺氧/复氧模型组(H/R组)、H/R+CPG-ODN组、H/R+PD-L1组,细胞计数试剂盒(CCK-8)检测各组细胞活性,乳酸脱氢酶(LDH)检测各组LDH活性,实时荧光定量聚合酶链反应检测各组中PD-L1、bcl-2、bax mRNA表达。采用单因素方差分析进行组间比较。结果I/RI后心脏组织中T细胞比例明显上升;Pdcd1基因[程序性死亡因子-1(PD-1)的靶基因]是I/RI后变异系数最大基因;CCK-8结果显示H/R+CPG-ODN组细胞存活率高于H/R和H/R+PD-L1组(73.88±3.40比55.24±2.17、45.89±1.99,F=988.50,P<0.01);LDH检测结果显示H/R组、H/R+CPG-ODN组、H/R+PD-L1组高于Control组(171.972±2.551、140.276±8.204、231.931±12.932比113.806±13.526,F=73.20,P<0.05);H/R+CPG-ODN组低于H/R组(140.276±8.204比171.972±2.551,t=7.88,P<0.01);H/R+PD-L1组高于H/R组(231.931±12.932比171.972±2.551,t=6.39,P<0.05);I/RI组IL-2、IL-6、IFN-γ高于假手术组(95.51±5.29、156.27±11.89、43.75±0.87比38.13±2.67、59.38±5.55、26.66±0.23,t=37.11、12.80、41.15,P<0.001);蛋白印迹法结果显示,CPG-ODN组PD-L1高于H/R组(3.06±0.38比1.79±0.11,t=5.66,P<0.05);CPG-ODN组bcl-2高于H/R组(0.95±0.09比0.66±0.02,t=5.18,P<0.05),PD-L1组bax高于H/R组(1.54±0.02比1.18±0.01,t=6.88,P<0.05);H/R+CPG-ODN组PD-L1相对表达量高于H/R组(4.39±0.45比2.15±0.28,t=7.32,P<0.05);H/R+CPG-ODN组bcl-2相对表达量高于H/R+PD-L1组(0.91±0.08比0.45±0.08,t=7.40,P<0.05);H/R+PD-L1组bax相对�ObjectiveTo investigate the effect of programmed death receptor 1(PD-L1)and its ligand signaling pathway on myocardial ischemia-reperfusion injury(I/RI)and the underlying mechanism.MethodsTotally,20 C57 BL mice aged 8-10 weeks were randomly divided into sham operation group(Sham group),I/RI group,I/RI+CPG-ODN group and I/RI+PD-L1 group,with 5 mice in each group.After modeling,the single cell sequencing was done.Serum interleukin(IL)-2,IL-6 and interferon-γ(IFN-γ)were determined.The protein expression levels of PD-L1,B cell lymphoma/leukemia-2(bcl-2)and bcl-2 associated X protein(bax)in tissues were detected by Western blotting.H9C2 cardiomyocytes were randomly divided into normal control group(control group),hypoxia/reoxygenation model group(H/R group),H/R+CPG-ODN group,H/R+PD-L1 group.Except for the control group,the H9C2 cells in the rest groups were pretreated with corresponding pretreatment for 30 min,and then treated with hypoxia for 3 h and reoxygenation for 3 h.The activity of H9C2 cells in each group was detected by Cell Counting Kit-8(CCK-8)method.The activity of LDH in H9C2 cells in each group was detected by LDH kit.The expression of PD-L1,bcl-2 and bax mRNA in H9C2 cells was detected by RT-PCR.One-way analysis of variance was used for comparison between groups.ResultsThe proportion of T lymphocytes in the heart tissue of C57BL mice increased significantly after I/RI.The Pdcd1 gene,which is the target gene of programmed death factor-1(PD-1),has the highest coefficient of variation after I/RI;CCK-8 assay showed that the cell viability in H/R+CPG-ODN group was higher than that in H/R and H/R+PD-L1 groups(73.88±3.40 vs.55.24±2.17,45.89±1.99,F=988.50,P<0.01).LDH levels in H/R group,H/R+CPG-ODN group and H/R+PD-L1 group were higher than in control group(171.972±2.551,140.276±8.204,231.931±12.932 vs.113.806±13.526,F=73.20,P<0.05),those in H/R+CPG-ODN group were lower than in H/R group(t=7.879,P<0.01),and those in H/R+PD-L1 group was higher than in H/R group(t=6.389,P<0.05).IL-2,IL-6 and IFN-γlevels

关 键 词：心肌缺血再灌注损伤 程序性死亡受体1及其配体信号通路 炎性因子 心肌保护 

分 类 号：R54[医药卫生—心血管疾病]