CD4^+T淋巴细胞Kv1.3通道在大鼠动脉粥样硬化中的作用

Role of Kv1. 3 channel of CD4^+ T lymphocytes in the formation of atherosclerosis in rat spleen

目的:探讨大鼠动脉粥样硬化(AS)模型中CD4+T淋巴细胞电压门控钾通道(Kv)Kv1.3的表达、功能及其在AS中的作用。方法:采用高脂饮食法建立大鼠动脉粥样硬化模型,流式细胞术分析淋巴细胞的比例,采用免疫磁珠法分离CD4+T淋巴细胞,研究脾组织CD4+T淋巴细胞Kv1.3mRNA表达、细胞内钙离子浓度及细胞因子分泌的变化。结果:(1)AS组脾组织CD4+T淋巴细胞占总T淋巴细胞比例较对照组明显升高(74.93%±2.15%vs67.80%±2.54%,P<0.05)。(2)经刀豆蛋白A(ConA)刺激,AS组T淋巴细胞增殖程度明显高于对照组(1.1321±0.1750vs0.7971±0.0955,P<0.05)。(3)AS组脾组织CD4+T淋巴细胞在ConA刺激状态下胞内钙离子浓度明显高于对照组(H=82,P<0.05)。(4)AS组脾组织CD4+T淋巴细胞在刺激48h后较刺激24h后细胞因子(IL-2,TNF-α)分泌显著增加。(5)AS组脾组织CD4+T淋巴细胞Kv1.3mRNA表达明显高于对照组(3.670±1.579vs1)。结论:AS组脾组织CD4+T淋巴细胞比例高于对照组,CD4+T淋巴细胞Kv1.3mRNA表达增多,提示高表达Kv1.3的CD4+T淋巴细胞可能在AS的发生发展中发挥重要的作用。

AIM： To investigate the function of voltage-gated potassium channel Kv1. 3 and its possible role in CD4 ＋ T lymphocytes in the formation of atherosclerosis （ AS） in rat spleen. METHODS： The rat atherosclerosis model was established by feeding high-fat diet. The proportion of lymphocytes was determined by flow cytometry. The CD4 ＋ T lymphocytes were separated using immunomagnetic bead. The mRNA expression of Kv1. 3 in CD4 ＋ T lymphocytes was detected. The concentrations of intracellular calcium and cytokines were also measured. RESULTS： （ 1） The proportion of CD4 ＋ T lymphocytes in AS group was significantly higher than that in control group （ 74. 93% ± 2. 15% vs 67. 80% ± 2. 54% ,P 0. 05） . （ 2） After stimulated with concanavalin A （ ConA） ,the proliferation of CD4 ＋ T lymphocytes in AS group was significantly higher than that in control group （ 1. 1321 ± 0. 1750 vs 0. 7971 ± 0. 0955,P 0. 05） . （ 3） After stimulated with ConA,the concentration of intracellular calcium in AS group was higher than that in control group. （ 4） In AS group,the releases of cytokines of IL-2 and TNF-α in AS group were significantly higher when stimulated with ConA for 48 h than that for 24 h. （ 5） The mRNA expression of Kv1. 3 in CD4 ＋ T lymphocytes was greatly higher in AS group than that in control group （ 3. 670 ± 1. 579 vs 1） . CONCLUSION： In AS rats,the increase in CD4 ＋ T lymphocytes as well as the augmentation of Kv1. 3 mRNA expression in the cells suggest that up-regulation of Kv1. 3 mRNA expression in CD4 ＋ T lymphocytes may be involved in the mechanism of atherosclerotic formation in rat spleen.