论文部分内容阅读
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CD4+ CD28null亚型活化前后IKCa1钾通道数目的变化以及IKCa1钾通道阻滞剂对活化CD4+ T细胞效应分子表达的影响,探讨IKCa1钾通道对不稳定斑块的意义。方法:用免疫磁珠法分离出24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 T细胞,采用全细胞膜片钳技术记录细胞活化前及活化3d后的IKCa1钾电流。CD4+T细胞活化3d后分别加入终浓度为0.2、1、5μmol/L特异性IKCa1钾通道阻滞剂TRAM-34,再继续活化3d,用反转录-PCR法检测干扰素-γ及颗粒酶B mRNA的表达。结果:活化后CD4+、CD4+ CD28 null T细胞的IKCa1通道数目分别增加了9倍和8倍[活化前后2种细胞的通道数分别为(45±3)∶(439±33),(56±4)∶(497±45),均P<0.01]。2种细胞的通道密度也分别增加了约3倍(P<0.01)。活化前及活化后2种细胞的通道数目及通道密度均无差别。不同浓度的TRAM-34均下调CD4+ T细胞活化后干扰素-γ、颗粒酶B mRNA的表达,各浓度组间干扰素-γ、颗粒酶B mRNA的表达差异均有统计学意义(P<0.01),浓度越高,各mRNA表达越低。结论:ACS患者外周血CD4+及CD4+ CD28null T细胞活化后IKCa1的通道数目及通道密度均明显增加,特异性IKCa1通道阻滞剂TRAM-34呈浓度依赖性地抑制CD4+T细胞活化时干扰素-γ及颗粒酶B mRNA的表达,提示CD4+T细胞的IKCa1钾通道可作为预防动脉粥样斑块不稳定的潜在治疗靶点。
Aims: To investigate the changes of IKCa1 potassium channels in peripheral blood of patients with acute coronary syndrome (ACS) before and after activation of CD4 + T cells and CD4 + CD28null subtypes and the effect of IKCa1 potassium channel blockers on the effector molecule expression of activated CD4 + T cells. Significance of IKCa1 Potassium Channel on Unstable Plaque. Methods: CD4 + T cells were isolated from peripheral blood of 24 patients with ACS by immunomagnetic beads method. 12 subtypes of CD4 + CD28null T cells were further subdivided. Whole cell patch clamp technique was used to record IKCa1 Potassium current. CD4 + T cells were activated after 3d were added to a final concentration of 0.2,1,5μmol / L specific IKCa1 potassium channel blockers TRAM-34, and then continue to activate 3d, reverse transcription-PCR assay interferon-γ and particles Enzyme B mRNA expression. Results: The numbers of IKCa1 channels in CD4 + and CD4 + CD28 null T cells increased 9-fold and 8-fold respectively after activation (45 ± 3 vs 43 ± 33 and 56 ± 4, respectively) ): (497 ± 45), all P <0.01]. The channel densities of the two kinds of cells also increased about 3-fold (P <0.01), respectively. There was no difference in the number of channels and the channel density between the two kinds of cells before and after activation. The expression of interferon-γ and granzyme B mRNA were decreased after different concentrations of TRAM-34 were activated in CD4 + T cells. The expression of IFN-γ and granzyme B mRNA in each concentration group had statistical significance (P <0.01 ), The higher the concentration, the lower the expression of each mRNA. CONCLUSIONS: The number of IKCa1 channels and the density of channels in peripheral blood CD4 + and CD4 + CD28null T cells of patients with ACS are significantly increased. The specific IKCa1 channel blocker TRAM-34 inhibits the activation of CD4 + T cells in a concentration-dependent manner. γ and granzyme B mRNA expression, suggesting that IKCa1 potassium channels in CD4 + T cells may serve as potential therapeutic targets for the prevention of atherosclerotic plaque instability.