依普利酮对慢性心力衰竭患者CD4+T淋巴细胞Kv1.3等通道mRNA及蛋白表达的抑制作用

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (11): 1259-1264.

依普利酮对慢性心力衰竭患者CD4⁺T淋巴细胞Kv1.3等通道mRNA及蛋白表达的抑制作用

李少华, 伊力哈木江·克尤木, 徐琦, 邵培培, 程路峰

¹新疆医科大学基础医学院药理学教研室，²第一附属医院心外科，³ 基础医学院免疫学教研室，乌鲁木齐 830011，新疆

收稿日期:2017-07-05 修回日期:2017-08-10 出版日期:2017-11-26 发布日期:2017-12-11
通讯作者: 程路峰，男，教授，硕士生导师，研究方向：心血管药理学、分子免疫学。 E-mail: lewis_clf@163.com
作者简介:李少华，在读硕士生，研究方向：心血管药理。 E-mail: lsh881212@163.com
基金资助:
国家自然科学基金（81360491）

Inhibitory effects of eplerenone on the mRNA and protein expression of Kv1.3 and other channels of CD4+T lymphocyte in patients with chronic heart failure

LI Shaohua ¹, YILI Hamujiang·KE Youmu², XU QI ³, SHAO Peipei ¹, CHENG Lufeng¹

¹Department of Pharmacology, School of Basic Medicine; ²Department of Cardiology, the First Affiliated Hospital; ³ Department of Immunology, School of Basic Medicine, Urumqi 830011, Xinjiang, China

Received:2017-07-05 Revised:2017-08-10 Online:2017-11-26 Published:2017-12-11

摘要/Abstract

摘要：

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。

关键词: 慢性心衰, CD4⁺T淋巴细胞, 依普利酮, Kv1.3通道, 抑制

Abstract:

AIM: To investigate the reversing effect of eplerenone on chronic heart failure through regulating the Kv1.3 channel of CD4+ T lymphocytes. METHODS: Thirty cases of chronic heart failure as CHF group and twenty cases of normal people as control group were collected from the First Affiliated Hospital of Xinjiang Medical University. CD4+ T lymphocyte was sorted by using immune magnetic bead which separated from peripheral blood, and its purity was tested by flow cytometry. The minimum effective concentration of EPL incubation CD4+ T lymphocyte proliferation was found by the CCK-8 technic; Control group, CHF group and CHF + EPL group were established. The expression of channel gene, Kv1.3, KCa3.1 and CRAC on the CD4+ T cell membrane were detected with RT-qPCR technique. And the protein expression of Kv1.3 channels on CD4+ T cell was detected by Western blot. RESULTS: 30 μmol/L was detedted as the prefect concentration for EPL. Compared with control group, the expression of mRNA and protein for Kv1.3 channel in CHF group were increased 10.74 times and 1.20 times, respectively (P<0.01), while that in the EPL+CHF group was decreased by 64.80% and 39.62%, respectively (P<0.01); the mRNA expressions of KCa3.1 and CRAC channel were also increased 4.73 times and 3.77 times, respectively in CHF group (P<0.01) but it decreased after exposed to EPL (decreased by 19.30% and 37.14%，P<0.01). CONCLUSION: Eplerenone can down-regulate the expression of Kv1.3, KCa3.1 and CRAC channel, especial the Kv1.3, which in accordance with higher expression of the three channels protein on the CD4+ T cell separated from CHF patients. And the aldosterone antagonist may inhibit the actibation of Kv1.3 membrane channel on the CD4+ T cell, which is beneficial for the CHF.

Key words: chronic heart failure, CD4+ T lymphocytes, eplerenone, Kv1.3 channel, inhibition

中图分类号:

R541

李少华, 伊力哈木江·克尤木, 徐琦, 邵培培, 程路峰. 依普利酮对慢性心力衰竭患者CD4⁺T淋巴细胞Kv1.3等通道mRNA及蛋白表达的抑制作用[J]. 中国临床药理学与治疗学, 2017, 22(11): 1259-1264.

LI Shaohua, YILI Hamujiang·KE Youmu, XU QI, SHAO Peipei, CHENG Lufeng. Inhibitory effects of eplerenone on the mRNA and protein expression of Kv1.3 and other channels of CD4+T lymphocyte in patients with chronic heart failure[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(11): 1259-1264.

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