CAJ | 학술논문

目的研究急性冠状动脉综合征(ACS)患者外周血中CD4+T细胞及CD28null/CD28+亚型活化前后Kv1.3钾通道数目的变化以及Kv1.3钾通道阻滞剂对CD4+T细胞活化表达的影响,探讨Kv1.3钾通道在不稳定斑块中的意义.方法用免疫磁珠法分离出27例ACS患者外周血中的CD4+T细胞,其中12例进一步分出亚型CD4+CD28null和CD4+CD28+T细胞,采用全细胞膜片钳技术记录细胞活化前及经CD3抗体活化72 h后的Kv1.3钾电流.CD4+T细胞活化时分别加入终浓度为0.1、1、10 nmol/L特异性Kv1.3钾通道阻滞剂rMargatoxin(rMgTX),共同培养72 h后用反转录-PER法检测干扰素-γ、肿瘤坏死因子(TNF)-α及颗粒酶B mRNA的表达.结果活化后CD4+、CD4+CD28null、CD4+CD28+T细胞的Kv1.3钾通道的峰电流均明显增加,细胞平均通道数分别增加约90%、60%、80%[活化前后每个细胞的通道数分别为(402±88)个比(752±275)个、(553±328)个比(874±400)个、(392±133)个比(716±251)个,均P<0.05].活化前CD4+CD28nullT细胞Kv1.3钾通道的平均数目比CD4+CD28+T细胞多约40%(P<0.05),活化后两者差异无统计学意义(P=0.102).不同浓度的rMgTX均下调CD4+T细胞活化后干扰素-γ、TNF-α、颗粒酶B mRNA的表达,各浓度组间干扰素-γ、TNF-α、颗粒酶B mRNA的表达差异均有统计学意义(均P<0.01),浓度越高,各mRNA表达越低.结论 ACS患者外周血CD4+T细胞及CD28null/CD28+亚型活化后Kv1.3钾通道表达增加,特异性Kv1.3通道阻滞剂rMgTX呈浓度依赖性地抑制CD4+T细胞活化时干扰素-γ、TNF-α及颗粒酶B mRNA的表达,提示CD4+T细胞特别是CD4+CD28nullT细胞的Kv1.3钾通道可作为预防动脉粥样斑块不稳定的潜在治疗靶点.
목적 연구급성관상동맥종합정(ACS)환자외주혈중CD4+T세포급CD28null/CD28+아형활화전후Kv1.3갑통도수목적 변화이급Kv1.3갑통도조체제대CD4+T세포활화표체적영향,탐토Kv1.3갑통도재불은정반괴중적의의.방법 용면역자주법분리출27례ACS환자외주혈중적CD4+T세포,기중12례진일보분출아형CD4+CD28null화CD4+CD28+T세포,채용전세포막편겸기술기록세포활화전급경CD3항체활화72 h후적Kv1.3갑전류.CD4+T세포활화시분별가입종농도위0.1、1、10 nmol/L특이성Kv1.3갑통도조체제rMargatoxin(rMgTX),공동배양72 h후용반전록-PER법검측간우소-γ、종류배사인자(TNF)-α급과립매B mRNA적표체.결과 활화후CD4+、CD4+CD28null、CD4+CD28+T세포적Kv1.3갑통도적봉전류균명현증가,세포평균통도수분별증가약90%、60%、80%[활화전후매개세포적통도수분별위(402±88)개비(752±275)개、(553±328)개비(874±400)개、(392±133)개비(716±251)개,균P<0.05].활화전CD4+CD28nullT세포Kv1.3갑통도적평균수목비CD4+CD28+T세포다약40%(P<0.05),활화후량자차이무통계학의의(P=0.102).불동농도적rMgTX균하조CD4+T세포활화후간우소-γ、TNF-α、과립매B mRNA적표체,각농도조간간우소-γ、TNF-α、과립매B mRNA적표체차이균유통계학의의(균P<0.01),농도월고,각mRNA표체월저.결론 ACS환자외주혈CD4+T세포급CD28null/CD28+아형활화후Kv1.3갑통도표체증가,특이성Kv1.3통도조체제rMgTX정농도의뢰성지억제CD4+T세포활화시간우소-γ、TNF-α급과립매B mRNA적표체,제시CD4+T세포특별시CD4+CD28nullT세포적Kv1.3갑통도가작위예방동맥죽양반괴불은정적잠재치료파점.
Objective To study the Kv1.3 channel expression changes after CD4 + and subsets CD28null/CD28+T cells activation in peripheral blood of patients with acute coronary syndrome (ACS).Methods CD4+ T cell in 27 ACS patients and CD4+ CD28null/CD4+ CD28 + T cells in 12 out of these 27 ACS patients were isolated from peripheral blood with magnetic cell sorting.The whole-cell Kv1.3 currents for three T cells were recorded with patch-clamp technique before and 72 hours after activation by purified anti-human CD3 Interferon gamma,tumor necrosis factor alpha(TNF-α),granzyme B mRNA expression were determined by reverse transcription-PCR before and 72 hours after activation by purified anti-human CD3 in the presence or absence of recombinant Margstoxin (rMgTX,0.1,1,10 nmol/L),a specific Kv1.3 channel blocker.Results Peak Kv1.3 channel currents of CD4 + ,CD4+ CD28null,CD4+ CD28 + T cells were significantly increased and the mean Kv1.3 channel numbers per cell of these cells were increased by about 90%,60%,80% (402 ± 88 vs.752 ± 275,553 ± 328 vs.874 ± 400,392 ± 133 vs.716 ± 251,all P <0.05) after activation compared to baseline values.Baseline CD4+ CD28nullT cell numbers were about 40% more than those of CD4+ CD28+ T cell (P < 0.05) and were similar after activation (P = 0.102).The mRNA expression of interferon gamma,TNF-α and granzyme B were dose-dependently down-regulated by rMgTX.Conclusions Kv1.3 channels of peripheral CD4 + T cell and CD28null/CD28 + T cells from ACS patients significantly increased after activation and Kv1.3-specific channel blocker rMgTX could effectively abolish this effect suggesting a potential role of Kv1.3 channel blocker on plaque stabilization in ACS patients.