The role of Kv1.3 channel of CD4+T lymphocytes in the formation of atherosclerosis in rat

目的：探讨大鼠动脉粥样硬化（Atherosclerosis，AS）模型中CD4+T淋巴细胞电压门控钾通道（Votage-gatedpotassiumchannel）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，通过RealtimePCR法研究脾组织CD4+T淋巴细胞膜上Kv1.3在mRNA水平的表达，激光共聚焦显微镜（Confocal）测定细胞内钙离子浓度，并通过酶联免疫吸附试验（ELISA）测定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...Objective：To investigate the function of voltage-gated potassium channel Kv1.3 and the possible role of its expression in CD4+ T lymphocytes in the formation of atherosclerosis(AS) in rat spleen. Methods : First, we established atherosclerosis model in rat with high-fat diet, then analyzed the proportion of lymphocytes by flow cytometry. By separating CD4+ T lymphocytes with immunomagnetic bead, ...学位：医学硕士院系专业：医学院临床医学系_内科学学号：2452007115252

自杀基因治疗恶性肿瘤的研究现状及展望

肿瘤的自杀基因疗法是近年来肿瘤基因治疗的研究热点,是一种具有潜在临床应用前景的新的肿瘤基因治疗策略。本文就近年来自杀基因疗法在肿瘤治疗中取得的研究进展,分别从作用机制、自杀基因系统、旁观者效应、自杀基因的转导以及联合基因治疗等几个方面进行综述

CD4~+T淋巴细胞Kv1.3通道在大鼠动脉粥样硬化中的作用

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

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.国家自然科学基金资助项目(No30871045

Expression profile of voltage-gated potassium channel Kv1.3 in aorta of atherosclerosis rats

目的探讨动脉粥样硬化(AS)大鼠主动脉病变局部离子通道kV1.3的表达水平及作用。方法雄性WISTAr大鼠16只,随机分为两组:正常组(8只,予普通饮食+0.9%氯化钠溶液)和AS组(8只,予高脂饮食+维生素d3负荷)。采用组织病理学检查,观察主动脉粥样硬化病变。采用实时定量反转录-聚合酶链反应(rT-PCr)和WESTErn印迹法检测主动脉病变局部kV1.3和白细胞介素(Il)-2、干扰素(Ifn)-γ的表达水平。结果组织病理学检查证实纤维增生性AS斑块。AS组的kV1.3MrnA为(31.48±8.64)x10-3,显著高于正常组的(3.28±0.79)x10-3(P=0.012)。AS组的kV1.3、Il-2、Ifn-γ蛋白表达量分别为0.691±0.067、0.611±0.055、0.759±0.050,均显著高于正常组的0.490±0.052、0.299±0.058、0.273±0.052(P值均<0.01)。结论 kV1.3在主动脉粥样硬化病变局部的表达增高。kV1.3可能在AS的发生和发展过程中发挥着重要的作用。Objective To investigate the expression of voltage-gated potassium channel Kv1.3 in the aorta of a rat model of atherosclerosis and its role in the progress of atherosclerotic plaque formation.Methods A total of 16 male Wistar rats were randomly divided into normal control(normal diet and saline)and atherosclerosis group(high lipid diet+Vitamin D3 overload),with 8 rats in each group.In 16 weeks later,all rats were killed after weighing,and their blood samples and aorta were collected.Pathological changes of the rat aortic artery were observed with HE staining.Real time RT-PCR and Western blotting analysis were used to determine the mRNA and protein expression of Kv1.3 and the protein expressions of interleukin(IL)-2 and interfereron(IFN)-γ.Results Pathological changes showed that fiber proliferative atherosclerotic plaques were found in the aorta of atherosclerosis group,with inflammatory cells infiltrating in the local lesion.Real time RT-PCR analysis showed that the expression of Kv1.3 mRNA in the aorta was increased significantly in the atherosclerosis rats than that in the controls([31.48±8.64]×10-3 vs.[3.28±0.79]×10-3,P<0.05).Western blotting analysis showed that the protein expression of Kv1.3,IL-2 and IFN-γ in the aorta were also increased significantly in the atherosclerotic rats than that in the controls(Kv1.3[0.691±0.067] vs.[0.490±0.052],IL-2 [0.611±0.055] vs.[0.299±0.058],IFN-γ [0.759±0.050] vs.[0.273±0.052],P<0.01 n=8).Conclusion The expression of Kv1.3 potassium channels is increased in the plaques of atherosclerotic rats.Kv1.3 may play an important role in the development and progression of atherosclerosis.国家自然科学基金资助项目(30871045