丝裂原活化蛋白激酶/细胞外调节蛋白激酶通路在动静脉内瘘术后内膜增生部位血管组织和细胞中的表达及意义  

Expression and significance of mitogen-activated protein kinase/extracellular signal-regulated kinase pathway in vascular tissue at site of intimal hyperplasia after arteriovenous fistula and myofibroblasts in uremic sera

作　　者：王自强 韦泽丰 郑金花 朱永俊[1] 程颖 吕潇阳 Wang Zi-qiang;Wei Ze-feng;Zheng Jin-hua;Zhu Yong-jun;Cheng Ying;Lyu Xiao-yang(Department of Nephrology,The First Affiliated Hospital of Hainan Medical University,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院肾内科,海口570102

出　　处：《临床肾脏病杂志》2024年第7期536-542,共7页Journal Of Clinical Nephrology

基　　金：海南省医药卫生科研项目(21A200026);海南医学院第一附属医院青年培育基金项目(HYYFYPY202109)。

摘　　要：目的探讨丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)/细胞外调节蛋白激酶(extracellular signal-regulated kinase,ERK)通路在动静脉内瘘术后内膜增生部位血管组织中的表达及意义,初步探索抑制MAPK/ERK通路对肌成纤维细胞自噬及增殖过程的影响。方法选取2023年1月至3月期间在海南医学院第一附属医院肾内科行动静脉内瘘重建术的患者6例,观察组收集动静脉内瘘重建术患者内膜增生严重部位的静脉血管组织,对照组为重建术中需结扎的侧支正常静脉组织。术前收集患者的一般资料,超声测量拟切取部位血管内径及内膜厚度,蛋白质印迹法检测血管组织中磷酸化细胞外调节蛋白激酶(phosphorylation-extracellular regulat-ed protein kinases 1/2,p-ERK1/2)、B细胞淋巴瘤/白血病-2蛋白相互作用蛋白1(myosin-like B cell lymphoma/lewkmia-2 interacting protein1,Beclin^(-1))、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)蛋白的表达水平,并分析其与静脉内膜厚度的相关性。以肌成纤维细胞为研究对象,尿毒症血清培养,并予ERK1/2抑制剂去氢钩藤碱(Corynoxeine)干预,提取细胞总蛋白及RNA,蛋白质印迹法及实时定量PCR检测Beclin^(-1)及PCNA的表达变化。结果入组患者透析过程中均存在血流量不足,灰阶超声提示动静脉内瘘狭窄处静脉内膜明显增厚[(0.143±0.014)cm比(0.097±0.016)cm],血管腔内径为(0.166±0.028)cm。蛋白质印迹法结果显示,与正常侧支部位血管组织相比较,内膜严重增生部位血管组织中p-ERK1/2、Beclin^(-1)、PCNA蛋白的表达明显增加,差异有统计学意义(均P<0.05)。观察组静脉内膜厚度与p-ERK1/2、Beclin^(-1)、PCNA蛋白的表达水平均呈正相关(R2=0.929、0.702、0.789,均P<0.05)。在细胞模型上,与正常对照组相比较,尿毒症血清组自噬相关指标Beclin^(-1)及增殖相关指标PCNA的表达明显升高,差异有统计学意义(均P<0.05)。而加入ERK特�Objective To explore the expression and significance of mitogen-activated protein kinase(MAPK)/extracellular signal-regulated kinase(ERK)pathway in vascular tissue of intimal hyperpla-sia after arteriovenous fistula(AVF)and explore the effect of suppressing MAPK/ERK pathway on au-tophagy and proliferation of myofibroblasts.Methods From January 2023 to March 2023,6 patients un-dergoing reconstruction of AVF were selected.Vascular tissues of patients with severe intimal hyperplasia were collected in observation group while normal collateral venous tissues requiring ligation in control group.General data were collected before operation and inner diameter and intima thickness of vessels were measured by ultrasound.The vascular expression levels of phosphorylation-extracellular regulated protein kinase 1/2(p-ERK1/2),myosin-like B cell lymphoma/leukemia-2 interacting protein 1(Beclin^(-1))and proliferating cell nuclear antigen(PCNA)proteins were detected by Western blot(WB)and correla-tion with venous intimal thickness was examined.Myofibroblast was cultured in uremic sera and exposed to ERK1/2 inhibitor corynoxeine.Total protein and RNA of cells were extracted and the expression changes of Beclin^(-1) and PCNA detected by WB and real-time polymerase chain reaction(PCR).Results Insufficient blood flow appeared during dialysis.Gray scale ultrasound indicated that venous in-tima thickened obviously[(0.143±0.014)cm vs(0.097±0.016)cm]at the stenosis of AVF and average di-ameter of vascular lumen was(0.166±0.028)cm.As compared with normal collateral vascular tissues,the expressions of p-ERK1/2,Beclin^(-1) and PCNA proteins were markedly up-regulated in vascular tissues of severe intimal hyperplasia and the differences were statistically significant(P<0.05).In observation group,venous intimal thickness was correlated positively with the expression levels of P-ERK1/2,Beclin^(-1) and PCNA proteins(R2 values of 0.929,0.702 and 0.789,P<0.05).As compared with normal control group,the expressions of Beclin^(-1) and PCNA spiked

关 键 词：丝裂原活化蛋白激酶 细胞外调节蛋白激酶 动静脉内瘘 内膜增生 

分 类 号：R692.5[医药卫生—泌尿科学]