芦丁对D-半乳糖诱导衰老小鼠认知功能的保护作用研究

Protective effect of rutin on cognitive function of D-galactose-induced aging mice

目的研究芦丁对D-半乳糖诱导衰老小鼠认知功能的保护作用机制。方法将80只雄性ICR小鼠采用直接抽取法分为对照组、D-半乳糖组、D-半乳糖+芦丁组和D-半乳糖+维生素E组,每组20只。D-半乳糖诱导小鼠"衰老"模型,同时给予芦丁口服6周。使用Morris水迷宫实验,记录小鼠的逃避潜伏期、游泳距离和游泳路径,试剂盒检测海马组织总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)和一氧化氮合酶(NOS)活性以及丙二醛(MDA)和一氧化氮(NO)含量。用甲酚紫染色观测海马神经元损伤情况,用微管相关蛋白-2(MAP-2)观测海马神经元树突完整性情况,用半胱天冬酶-3(Caspase-3)、胶质纤维酸性蛋白(GFAP)抗体标记脑冠状切片来分别观测海马神经元凋亡情况与星型胶质细胞损伤情况。结果 D-半乳糖明显诱导小鼠海马区的神经毒性。与D-半乳糖组小鼠比较,D-半乳糖+芦丁组小鼠在第5、6天训练的逃避潜伏期和游泳距离均缩短(P<0.05),训练结束后实验结果显示,与D-半乳糖组小鼠比较,D-半乳糖+芦丁组小鼠海马完整神经元数增加(P<0.05),凋亡阳性细胞数明显减少(P<0.01),神经元树突损伤减轻,T-SOD和GSH-Px的异常活性恢复(均P<0.05),MDA水平降低(P<0.01)。此外,D-半乳糖+芦丁组小鼠中海马星形胶质细胞树突损伤有所缓解(P<0.05),细胞中NOS活性恢复,NO释放明显降低(均P<0.01)。而维生素E作为阳性对照有与芦丁相似的神经保护功能。结论芦丁可通过缓解脑损伤来改善D-半乳糖诱导衰老小鼠的认知功能,在脑老化过程中可能有正效应的作用。

Objective To study the protective mechanism of rutin on cognitive function in D-galactose(D-gal)-induced aging mice.Methods D-gal induced a"aging"model in mice.At the same time given rutin for 6 weeks.The Morris water maze(MWM)was used to assess spatial learning and memory in aging mice.The kit detects total superoxide dismutase(T-SOD),glutathione peroxidase(GSH-Px)and Nitric oxide synthase(NOS)activity and malondialdehyde(MDA)and nitric oxide(NO)levels.Coronal sections of the brain were labeled with cresyl violet or with microtubule-associated protein-2(MAP-2),caspase-3 and glial fibrillary acidic protein(GFAP)antibodies.Results Rutin significantly reduced the escape latency and swimming distance of D-gal treated mice,prevented hippocampal neuron loss and apoptosis induced by D-gal,and restored T-SOD and GSH-Px in the hippocampus of aging mice and reduce the level of MDA.In addition,rutin could alleviate dendritic damage of hippocampal astrocytes,restore NOS activity,and inhibit the NO release of cells.Conclusion Rutin can imprae the cognitive function of D-galactose-induced aging mice by relieving main damage,playing a protective role in brain aging.