动脉粥样硬化患者CD4^+CD25^+调节性T细胞上Kv1.3通道的活性变化  

Inhibitory effects of eplerenone on Kv1.3 and other channels of CD4^+CD25^+Treg in patients with atherosclerotic cardiovascular disease

作　　者：邵劲松 胡锴 王红瑞 王立伟 徐博 Shao Jinsong;Hu Kai;Wang Hongrui;Wang Liwei;Xu Bo(Department of Cardiovascular Surgery,First Affiliated Hospital of Air Force Medical University,Shaanxi xi'an 710032,China)

机构地区：[1]空军军医大学基础医学系学员六大队,西安710032 [2]石家庄飞行学院第三训练旅单位,运城市044500 [3]解放军空军军医大学第一附属医院心血管外科,西安710032

出　　处：《中国体外循环杂志》2020年第4期233-236,256,共5页Chinese Journal of Extracorporeal Circulation

摘　　要：目的探讨动脉粥样硬化性心血管疾病(ACVD)患者CD4^+CD25^+调节性T细胞(Tregs)的Kv1.3通道活性与该疾病的关系,并验证依普利酮(EPL)对动脉血管粥样硬化有逆转作用。方法收集ACVD患者(n=70)及正常人志愿者(n=20)全血样本。免疫磁珠分选外周血CD4^+CD25^+Tregs,CCK-8技术检测ELP对CD4^+CD25^+Tregs增殖抑制的影响。分组进行培养:正常人外周血的Control组、ACVD患者外周血组(ACVD组)、ACVD患者+EPL组(ACVD+EPL组)。RT-qPCR检测各组培养体系中的Kv1.3(钾通道)、KCa3.1(钙通道)和内质网钙释放激活的钙通道(CRAC)基因的表达;In-cell western blotting技术检测CD4^+CD25^+Tregs的Kv1.3通道蛋白质水平的表达。结果EPL的最适浓度为30μmol;ACVD组Kv1.3通道的mRNA和蛋白质的表达分别是正常对照组3.35倍和1.83倍(P<0.01),ACVD+EPL组Kv1.3通道的mRNA和蛋白质的相对表达分别是ACVD组62.09%(P<0.01);ACVD组KCa 3.1和CRAC通道mRNA的表达分别是正常对照组的2.18、2.22倍(P<0.01),ACVD+EPL组KCa 3.1和CRAC通道mRNA的表达是ACVD组的61.01%、76.68%(P<0.01)。结论CD4^+CD25^+Treg细胞的三种离子通道在ACVD中均有高表达,EPL可抑制Kv1.3通道的活性,推测醛固酮受体拮抗剂可能通过抑制CD4^+CD25^+Tregs上的Kv1.3通道的激活,抑制了T淋巴细胞的活性,以抑制动脉粥样硬化病变中炎症作用的发生发展。Objective To investigate the effect of Kv1.3 channel expression on the activity of CD4^+CD25^+Tregs in patients with atherosclerotic cardiovascular disease(ACVD),and to verify the reversal effect of eplerenone.Methods Blood samples of 70 cases of atherosclerotic cardiovascular disease as ACVD group and 20 cases of normal people as control group were collected in our hospital.CD4^+CD25^+Tregs separated from peripheral blood were sorted by immune magnetic bead,and the purity were tested by flow cytometry.The minimum effective concentration of EPL incubation on the proliferation of CD4^+CD25^+Tregs was found by CCK-8 technique.RT-qPCR technique was used to detection the expression of channel gene,such as Kv1.3,KCa3.1 and CRAC on the membrane of CD4^+CD25^+Tregs.ICW technique was used to detection the protein expression of Kv1.3 channels on CD4^+CD25^+Tregs.Results Compared with normal group,ACVD group showed higher expression of mRNA for Kv1.3 channel,KCa3.1 channel and CRAC channel(P<0.01),as well as higher expression of protein for Kv1.3 channel(P<0.01).Compared with ACVD group,EPL+ACVD group showed lower expression for Kv1.3 channel(P<0.01),both in gene and protein.Conclusion The three ion channels of CD4^+CD25^+Tregs are highly expressed in ACVD.EPL can inhibit the activity of Kv1.3 channel.It is speculated that aldosterone receptor antagonists may inhibit the activation of Kv1.3 channel on CD4^+CD25^+Tregs.Inhibiting the activity of T lymphocytes can inhibit the occurrence and development of inflammation in ACVD.

关 键 词：动脉粥样硬化性心血管疾病 CD4^+CD25^+Tregs Kv1.3通道 抑制 依普利酮 

分 类 号：R543.5[医药卫生—心血管疾病]