Laboratory of Physiology, Faculty of Nutrition, and Cooperative Research Center of Life Sciences, Kobe Gakuin University, Kobe, Japan.
Thromb Res. 2010 Mar;125(3):246-52. doi: 10.1016/j.thromres.2009.11.008. Epub 2009 Dec 16.
We have investigated the effects of differential aspirin doses on atherogenesis. Aspirin was given to homozygous, apoE(-/-) and LDLR(-/-) double deficient mice for 12 weeks. The development of arteriosclerosis was determined morphologically by image analysis and endothelial cell function was assessed by measurement of peripheral nitric oxide (NO).
ApoE(-/-) LDLR(-/-)double knockout mice were bred and maintained with a high fat diet containing aspirin (4 and 40 mg/kg B.W. /day) for twelve weeks. The development of arteriosclerosis was monitored by estimating the total area of atherosclerotic lesions in the entire aorta. Acetylcholine-induced NO release was measured in vivo using electrochemical sensors. The expression of eNOS on the endothelial surface was determined by immuno-staining. Plasma prostaglandin F1alpha (PGF(1 alpha)), serum thromboxian B(2) (TXB(2)) and total cholesterol were measured using enzymatic assay. Bleeding time was measured by tail cut method.
Arteriosclerosis in the 4 mg/kg/day aspirin group was decreased significantly compared with the placebo group, but not in the 40 mg/kg/day aspirin group. Acetylcholine-induced NO release was significantly depressed in the 40 mg/kg/day aspirin group. Immunochemical analysis with anti-eNOS antibody supported these findings. In the 4 mg/kg/day aspirin group, the severe suppression of PGI(2) production was not confirmed in spite of decreasing TXB(2) production, but not in the 40 mg/kg/day aspirin group.
Our results suggest that endothelial dysfunction with low dose aspirin improved, reduced progression of atherosclerosis in apoE(-/-) and LDLR(-/-) double deficient mice and provides a pathophysiological basis for the beneficial effects of aspirin in atherosclerosis, and low doses appeared to be more efficient than high doses.
我们研究了不同剂量阿司匹林对动脉粥样硬化形成的影响。将阿司匹林给予纯合子、载脂蛋白 E(-/-)和 LDLR(-/-)双基因敲除小鼠，连续给药 12 周。通过图像分析确定动脉硬化的发展，通过测量外周一氧化氮 (NO) 来评估内皮细胞功能。
apoE(-/-)LDLR(-/-)双基因敲除小鼠用高脂饮食喂养，并补充阿司匹林（4 和 40mg/kgBW/天）12 周。通过估计整个主动脉粥样硬化病变的总面积来监测动脉硬化的发展。使用电化学传感器在体内测量乙酰胆碱诱导的 NO 释放。通过免疫染色确定内皮表面的 eNOS 表达。使用酶联免疫吸附试验测定血浆前列腺素 F1alpha（PGF1alpha）、血清血栓素 B2（TXB2）和总胆固醇。用尾切法测量出血时间。
与安慰剂组相比，4mg/kg/天阿司匹林组的动脉粥样硬化明显减少，但 40mg/kg/天阿司匹林组无变化。40mg/kg/天阿司匹林组乙酰胆碱诱导的 NO 释放明显减少。用抗 eNOS 抗体进行免疫化学分析支持这些发现。在 4mg/kg/天阿司匹林组，尽管 TXB2 生成减少，但未确认 PGI2 生成的严重抑制，但在 40mg/kg/天阿司匹林组则有抑制。
我们的结果表明，低剂量阿司匹林改善了内皮功能障碍，减少了 apoE(-/-)和 LDLR(-/-)双基因敲除小鼠的动脉粥样硬化进展，并为阿司匹林在动脉粥样硬化中的有益作用提供了病理生理学基础，低剂量似乎比高剂量更有效。
