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目的:探讨依普利酮（EPL）对慢性心力衰竭（CHF）患者CD4+T淋巴细胞Kv1.3等通道的mRNA及蛋白质表达的影响。方法: 收集慢性心衰Ⅱ、Ⅲ期的患者及正常人全血样本（30例vs 20例）。免疫磁珠分选外周血CD4+T淋巴细胞，利用流式细胞仪检测其纯度，CCK-8法检测不同浓度的EPL对CD4+ T淋巴细胞增殖的影响。共分为3组，Control组、CHF组及CHF+EPL组。RT-qPCR检测各组培养体系中的Kv1.3、KCa3.1和CRAC通道mRNA的表达；In-cell Western blot技术检测CD4+ T淋巴细胞膜上Kv1.3通道蛋白质的表达。结果:EPL的最佳抑制浓度为30 μmol/L；CHF组Kv1.3通道的mRNA和蛋白质的表达分别是Control组的10.74倍和1.20倍（P<0.01），CHF+EPL组Kv1.3通道的mRNA和蛋白质的表达分别比CHF组降低了64.80%和39.62%（P<0.01）；CHF组KCa3.1和CRAC通道mRNA的表达分别是Control组的4.73倍和3.77倍（P<0.01），CHF+EPL组KCa3.1和CRAC通道mRNA的表达比CHF组分别降低了19.30%和37.14%（P<0.01）。结论:CD4+T淋巴细胞的三种离子通道的mRNA和/或蛋白质在慢性心力衰竭情况下均有高表达，依普利酮均能明显下调三种离子通道的mRNA和/或蛋白质，其中Kv1.3通道的变化尤为突出，推测醛固酮受体拮抗剂可能通过抑制CD4+T淋巴细胞膜上的Kv1.3通道的激活，减少T淋巴细胞的活化/增殖，最终抑制CHF过程中炎症作用的发生发展而对心衰有利。
Objective: To investigate the effects of Eplerenone (EPL) on mRNA and protein expression of Kv1.3 and other channels in CD4 + T lymphocytes in patients with chronic heart failure (CHF). METHODS: Whole blood samples from patients with chronic heart failure in stage II and III and in normal subjects (30 vs. 20) were collected. Peripheral blood CD4 + T lymphocytes were sorted by immunomagnetic beads. The purity of CD4 + T lymphocytes was detected by flow cytometry. The proliferation of CD4 + T lymphocytes was detected by CCK-8 assay. Divided into 3 groups, Control group, CHF group and CHF + EPL group. The expression of Kv1.3, KCa3.1 and CRAC channel mRNA in each culture system was detected by RT-qPCR. The protein expression of Kv1.3 channel in CD4 + T lymphocyte membrane was detected by In-cell Western blot. Results: The optimal inhibitory concentration of EPL was 30 μmol / L. The mRNA and protein expressions of Kv1.3 channel in CHF group were 10.74 and 1.20 times higher than those in control group (P <0.01) (P <0.01). The mRNA expression of KCa3.1 and CRAC in CHF group were 4.73 and 3.77 times (P <0.01) lower than that in CHF group The mRNA expression of KCa3.1 and CRAC in CHF + EPL group decreased by 19.30% and 37.14% respectively compared with CHF group (P <0.01). CONCLUSION: mRNA and / or protein of the three ion channels of CD4 + T lymphocytes are highly expressed in chronic heart failure cases. Eplerenone can obviously down-regulate the mRNA and / or protein of the three ion channels, of which Kv1 .3 channel changes are particularly prominent, presumed that aldosterone receptor antagonist may inhibit the activation of Kv1.3 channel CD4 (superscript +) T lymphocyte membrane, reduce the activation / proliferation of T lymphocytes, and ultimately inhibit the development of inflammatory processes in the development of CHF Heart failure and good.