D-半乳糖诱导衰老小鼠模型的建立与评价

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摘要 背景：目前构建衰老动物模型的方法主要有注射 D-半乳糖,臭氧损伤,去胸腺,采用老龄动物等。因 D-半乳糖造模简单、试验 周期相对较短，故目前广泛应用于衰老试验研究。 目的：使用 D-半乳糖建立亚急性衰老小鼠模型，并对其进行行为学、生化以及免疫等方面的评价。 设计、时间及地点：随机对照动物实验，于 2008-03/08 在南通大学医学院完成。 材料：ICR 小鼠 35 只，体质量(18±2) g，雌雄兼用。随机分为正常对照组 15 只，衰老模型组 20 只。 方法：衰老模型组小鼠用 D-半乳糖按 120 mg/kg 背部皮下注射 6 周，1 次/d，造成亚急性衰老模型；正常对照组不予药物。 主要观察指标：使用 Y 型电迷宫检测小鼠的学习、记忆功能；使用相关试剂盒测定血清中丙二醛浓度、超氧化物歧化酶活 力和脑中脂褐质含量、单胺氧化酶-B 活性以及肝中谷胱甘肽过氧化物酶活性等生化指标；单向琼脂扩散法测定血清中 IgG 含量；四甲基偶氮唑盐法测定小鼠脾淋巴细胞转化刺激指数；ELISA 检测血清中细胞因子白细胞介素 2，γ干扰素的浓度。 结果：与正常对照组相比，Y 迷宫显示衰老模型组小鼠记忆功能明显下降，表现为不同时间的正确反应率降低，潜伏期延 长；生化指标显示模型小鼠血清中丙二醛及脑中脂褐质含量增加、血清中超氧化物歧化酶与肝中谷胱甘肽过氧化物酶活力 下降，而脑中单胺氧化酶 B 活力没有明显改变；免疫相关指标显示模型小鼠血清 IgG 水平下降、脾淋巴细胞转化刺激指数 降低、血清中细胞因子γ干扰素的浓度减少，白细胞介素 2 变化不明显。 结论：D-半乳糖建立亚急性衰老模型小鼠记忆功能下降、氧自由基产物增加和抗氧化能力下降、免疫功能降低，可用于行 为学、抗氧化、以及免疫等方面的抗衰老药效研究。 关键词:D-半乳糖；衰老模型；免疫功能；行为；氧自由基；抗氧化；小鼠
ISSN 1673-8225 CN 21-1539/R CODEN: ZLKHAH
1275
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秦红兵，等. D-半乳糖诱导衰老小鼠模型的建立与评价
0 引言
D-半乳糖衰老动物模型是由国内学者龚国清等在 1991年根据衰老的代谢紊乱学说而构建的。与其他几种 衰老动物模型如SAMP系小鼠衰老模型、臭氧损伤衰老 模型、去胸腺衰老模型、自然衰老模型等相比，该模型 简便易行，价格低廉，结果稳定，因而广泛应用于药物 抗衰老的研究之中[1-2]。但是D-半乳糖诱导衰老模型在 行为、免疫等方面的衰老研究中的应用，文献相关报道 并不完全一致[3-5]。实验使用D-半乳糖建立亚急性衰老 小鼠模型，并对其进行行为学、生化以及免疫等方面的 评价。
秦红兵，杨朝晔，范忆江，朱清. D-半乳糖诱导衰老小鼠模型的建立与评价[J].中国组织工程研究与临床康复，2009，
13(7):1275-1278
[http://www.crter.org http://cn.zglckf.com]
1Yancheng Health Vocational and Technical College, Yancheng 224006, Jiangsu Province, China; 2Department of Pharmacology, Medical College of Nantong University, Nantong 226001, Jiangsu Province, China
秦红兵1，杨朝晔1，范忆江1，朱 清2
Establishment and evaluation of aging models induced by D-galactose in mice
Qin Hong-bing1, Yang Zhao-ye1, Fan Yi-jiang1, Zhu Qing2
Abstract BACKGROUND: At present, an aging model is mainly constructed by injection of D-galactose, ozone injury, removing thymus or using senile animals. D-galactose induced aging model is widely applied for aging research because of its simpleness and quickness. OBJECTIVE: To establish aging model produced with D-galactose in mice, and evaluate the behavior, biochemical index and immunologic function. DESIGN, TIME AND SETTING: Randomized controlled animal experiment, was accomplished in the Medical College of Nantong University from March to August 2008. MATERIALS: Thirty-five ICR mice, weighing (18±2) g, with both genders, were randomly divided into the normal control group (n =15) and aging model group (n=20). METHODS: Mice were given subcutaneously injection of D-galactose with 120 mg/kg per day for six weeks to induce subacute aging model. There was no medication in the normal control group. MAIN OUTCOME MEASURES: Y electric maze was used to determine the function of learning and memory. The analytic kit was used to measure the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in serum of mice, as well as the lipofuscin content and monoamine oxidase-B (MAO-B) activity in brain and glutathione peroxidase (GSH-PX) in liver. The single agar immunodiffusion method was used to detect IgG level in serum. Spleen lymphocyte transforming stimulation index was determined by methyl thiazolyl tetrazolium (MTT) colorimetric method. The level of Interleukin-2 (IL-2) and interferon-γ (IFN-γ) in serum of mice were determined by ELISA method. RESULTS: Compared with the normal control group, the results of Y-maze test showed that memory in the aging model mice was obvious reduced, the correct response rate was decreased with prolonged latency. The activity of SOD and GSH-PX decreased and the content of MDA and lipofuscin increased, however, there was no change in MAO-B activity. The IgG level of serum and the lymphocyte transforming stimulation index of spleen were declined, and the level of IFN-γ in serum was reduced, IL-2 did not change significantly. CONCLUSION: D-galactose induced subacute aging model, with declined memory ability, increased production of oxygen free radical, decreased antioxidative capacity, and reduced immunological function, can be used for immunological, antioxidative and behavioral research in aging.
Received: 2008-09-20 Accepted: 2008-12-06
1 盐城卫生职业技 术学院，江苏省盐 城市 224006； 2 南通大学医学院 药理学教研室，江 苏省南通市 226001
秦红兵，男，1959 年生，江苏省大丰 市人，汉族，1982 年南通医学院毕 业，副教授，主要 从事中药抗衰老 研究。 ywqhb2005@ 126.com
Qin HB, Yang ZY, Fan YJ, Zhu Q. Establishment and evaluation of aging models induced by D-galactose in mice.Zhongguo
Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2009;13(7): 1275-1278(China)
中国组织工程研究与临床康复 第 13 卷 第 7 期 2009–02–12 出版 Journal of Clinical Rehabilitative Tissue Engineering Research February 12, 2009 Vol.13, No.7
基础医学
D-半乳糖诱导衰老小鼠模型的建立与评价
通讯作者：朱 清，博士，讲师， 南通大学医学院 药理学教研室，江 苏省南通市 226001 zhuqing7883@ 163.com
中图分类号:R332 文献标识码:B 文章编号:1673-8225 (2009)07-01275-04
收稿日期：2008-09-20 修回日期：2008-12-06 (54200809190025/ W·Z)
Qin Hong-bing, Associate professor, Yancheng Health Vocational and Technical College, Yancheng 224006, Jiangsu Province, China ywqhb2005@ 126.com
Correspondence to: Zhu Qing, Doctor, Lecturer, Department of Pharmacology, Medical College of Nantong University, Nantong 226001, Jiangsu Province, China zhuqing7883@163. com