补肾祛痰化瘀法对载脂蛋白E基因敲除小鼠氧化应激水平的影响研究

摘要

Objective To assess the level of oxidative stress and to explore the effects of tonifying kidney and removing stasis plus reducing phlegm on oxidative stress of ApoE knockout mice in senior and elder groups. Methods ApoE knockout mice have been divided into four groups as the treatment group and model group of senior and the treatment group and model group of elder. And C57BL/6J mice have been divided into two groups as the normal groups in senior and elder. When the senior group mice were 28 weeks and the elders were 44 weeks, the mice brain tissue were tested by SOD, GSH-Px and MDA. Ob-serve the shape and number of the nerve cells in the hippocampus area by toluidine blue staining. And test the expression of 8- hydroxy deoxyguanosine by immunohistochemical method. Results The treatment group and model group and normal group in senior have no differences. But in elder group, the levels of ox-idative stress in treatment group and normal group have no difference with each other, but both have differ-ence with model group. Conclusion ApoE knockout mice had more oxidative damage when they get old. The method of tonifying kidney and removing stasis and reducing phlegm can improve oxidative damage on ApoE knockout mice.%目的：评估不同周龄载脂蛋白E( Apolipoprotein E,ApoE)基因敲除小鼠的氧化应激水平,并探讨补肾祛痰化瘀法对ApoE基因敲除小鼠氧化应激水平的影响。方法将成年组及老年组ApoE基因敲除小鼠共32只随机分为中药组及对照组,并取相同遗传背景的C57BL/6J小鼠共16只随机分成成年正常组及老年正常组,连续给予补肾祛痰化瘀法组方灌胃12周,成年组至28周龄、老年组至44周龄时予检测脑组织超氧化物歧化酶( superoxide dismutase, SOD)、谷胱甘肽过氧化物酶( glutathione peroxidase,GSH-Px)活力,及丙二醛( malondialdehyde, MDA)水平；并进行甲苯胺蓝染色观察海马CA1区神经细胞分布情况及尼氏体数量,免疫组化法检测8-羟基脱氧鸟苷(Oxo-2’-deox-yguanosine ,8-OHDG)表达情况。结果氧化应激水平均符合正态分布,故采用单因素方差分析进行数据统计。 ApoE基因敲除小鼠及C57小鼠28周龄时脑内氧化应激水平无明显差异；随着小鼠的周龄的增加,至44周时,老年ApoE基因敲除小鼠对照组其脑内MDA及8-OHDG表达水平明显增高,而SOD及GSH-Px表达水平明显降低,与老年正常组C57小鼠的脑内氧化应激水平具有明显差异(P<0．05)；而老年中药组与老年正常组相比,无明显差异。结论老年ApoE基因敲除小鼠氧化应激水平较高,而补肾祛痰化瘀法具有一定的抗氧化作用,可抵抗部分机体氧化作用,并控制机体氧化应激水平。