大鼠动脉粥样硬化斑块中T淋巴细胞亚群及Kv1.3通道蛋白的表达

目的探究Kv1.3通道蛋白与动脉粥样硬化(AS)模型中活化的T淋巴细胞间的关系。方法 Wistar雄性大鼠24只,随机分为对照组(n=10)和AS组(n=14),采用高脂饲料喂养方法建立AS模型。分别于实验开始前,实验第8周,实验第12周观察各组大鼠体重变化。于第12周处死大鼠前取血,检测血清中总胆固醇(TC)、低密度脂蛋白(LDL-L)、高密度脂蛋白(HDL-L)和甘油三酯(TG)的水平。通过病理HE染色及免疫组织化学方法观察AS斑块内T淋巴细胞亚群分布和Kv1.3通道蛋白表达的改变。结果 AS组体重、TC、LDL-C较对照组明显升高(P均<0.05);HDL-C和TG两组无差异。AS组主动脉管壁可见明显斑块形成,对照组血管壁各层的组织结构正常。AS组动脉斑块部位内膜下及中膜层可见CD4+与CD8+T淋巴细胞聚集,以CD4+T淋巴细胞聚集为主,在病变部位Kv1.3通道蛋白表达增加。对照组血管内膜、中膜中未见T淋巴细胞聚集及KV1.3通道蛋白的表达。结论 Kv1.3通道可能在调节AS斑块部T淋巴细胞亚群的激活中起着重要作用。

The voltage-gated potassium Kv1.3 channel expression changes in the subsets T lymphocyte in atherosclerotic plaque of Wistar rats

Objective To explore the relationship between Kv1.3 ion channel protein and activated T lymphocyte in atherosclerotic （AS） plaque. Methods Twenty-four male Wistar rats were randomly assigned to control group （n = 10） and AS model group（ n = 14）. AS model rats were established after being fed with high-cholesterol diet and vitamin D3 for 12 weeks, and the rats were weighted at the begining of the experiment as well as the 8th and 12th weeks. Before killed at the 12th weeks, the rats＇ blood was taken for trigliceride（TG） ,total cholesterol（TC） ,low and high density lipoprotein level （LDL-L, HDL-L） testing. Then rats were scarified, their aorta were isolated and dyed with hematoxylin-eosin for immun ohistochemical analysis. By using Image-Pro Plus system, the mean Integrated option density （IOD） of the area of interesting （AOI） in aorta were quantified. Results ①The weight of AS group was significantly higher than that of the control group;② The concentration of TC, LDL-L in AS group was significantly higher than that in control group （ P 〈 0.05 ）, while the HDL-L was higher （P 〉 0.05 ）, the TG was lower（ P 〉 0.05 ） ; ③ The aortic intima was found thicker in the AS group, proliferation was found in medial smooth muscle cell and was less orderly arranged, dot-flake calcification was observed in some specimen in AS group, while the morphology of aorta was relatively normal and cells were relatively orderly arranged in control group, Inflammatory cell infiltration and smooth muscle cell proliferating were not found in control group. In AS group, CD4 ^＋ and CD8^ ＋ T lymphocyte tended to aggregate in subintima and media of aorta. In the same area of AS plaque, CD4 ^＋ T lymphocyte were found much more, and Kv1.3 ion channel expression was quite positive in the same area, which was brown, and showed dot-flake distribution. T lymphocyte and Kv1.3 ion channel protein expression were negative in the aorta of control group. Conclusions Kv1.3 channel might play a role in the activation of T lympho cyte in atheromatous plaque, which possibly accelerates the development of atherosclerosis in rat.