天芪益智颗粒对Aβ1-42诱导的阿尔茨海默病大鼠学习记忆能力及蛋白质组的影响
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摘要
目的观察天芪益智颗粒对阿尔茨海默病(Alzheimer disease,AD)大鼠学习记忆能力和皮层、海马组织蛋白质组的影响,探讨其分子作用机制。方法将实验大鼠随机分为假手术组、模型组、阳性药组和天芪益智颗粒高、中、低剂量组,每组12只。灌胃给药,每日1次,连续40 d。给药结束后,采用Morris水迷宫实验评价大鼠学习记忆能力,应用TMT蛋白质组学方法进行质谱分析,确定天芪益智颗粒组与模型组间的差异蛋白质,应用PANTHER及STRING软件对差异蛋白进行生物信息学分析,采用Western blot对苹果酸脱氢酶和Fas相关的死亡结构域蛋白的表达变化进行验证。结果水迷宫实验结果显示,与模型组比较,天芪益智颗粒高、中剂量组定位巡航实验中逃避潜伏期缩短,空间探索实验中穿越目标区域次数显著增加(P<0.05,P<0.01);蛋白质组学结果显示,天芪益智颗粒高剂量组中成功鉴定33个差异蛋白质,这些蛋白及网络的功能涉及能量代谢、神经传递、细胞凋亡、细胞骨架、神经营养等,且Western blot蛋白表达检测结果与蛋白质组学结果一致。结论天芪益智颗粒可明显改善AD大鼠学习记忆能力,其机制与调节能量代谢、神经传递、细胞凋亡、细胞骨架及神经营养蛋白的表达密切相关。
Objective To observe the effects of Tianqi Yizhi Granules on learning and memory ability and proteomics in cerebral cortex and hippocampus of AD rats; To explore the molecular mechanism of Tianqi Yizhi Granules. Methods SD rats were randomly divided into sham-operation group, model group, Tianqi Yizhi Granules high-, medium-, and low-dosage groups, with 12 rats in each group. The rats received gavage for medication, once a day for 40 d. After the end of the administration, Morris water maze test was used to evaluate the effects of medicine on the learning and memory ability of the rats. The TMT proteomics method was used for mass spectrometry to determine the differential proteins between Tianqi Yizhi Granules groups and model group. Bioinformatics analysis of differential proteins was performed by using PANTHER and STRING software, and the expression changes of malate dehydrogenase and Fas-associated death domain protein were verified by Western blot. Results Morris water maze experiment showed that the escape latency of Tianqi Yizhi Granules high-and medium-dosage groups were shortened and the times of crossing target quadrant in space exploration experiment significantly increased(P<0.05, P<0.01)compared with model group. Proteomics results showed that there were 33 differentially expressed proteins between model and Tianqi Yizhi Granules groups. The functions of these proteins and protein network were related to energy metabolism, neurotransmission, cell apoptosis, cytoskeleton and neurotrophy. The Western blot results of malate dehydrogenase and FAS-associated death domain protein were consistent with proteomics results. Conclusion Tianqi Yizhi Granules can improve learning and memory ability of AD rats and the action of mechanism is closely related to the regulation of protein expression of energy metabolism, neurotransmission, apoptosis, cytoskeleton and neurotrophin.
Objective To observe the effects of Tianqi Yizhi Granules on learning and memory ability and proteomics in cerebral cortex and hippocampus of AD rats; To explore the molecular mechanism of Tianqi Yizhi Granules. Methods SD rats were randomly divided into sham-operation group, model group, Tianqi Yizhi Granules high-, medium-, and low-dosage groups, with 12 rats in each group. The rats received gavage for medication, once a day for 40 d. After the end of the administration, Morris water maze test was used to evaluate the effects of medicine on the learning and memory ability of the rats. The TMT proteomics method was used for mass spectrometry to determine the differential proteins between Tianqi Yizhi Granules groups and model group. Bioinformatics analysis of differential proteins was performed by using PANTHER and STRING software, and the expression changes of malate dehydrogenase and Fas-associated death domain protein were verified by Western blot. Results Morris water maze experiment showed that the escape latency of Tianqi Yizhi Granules high-and medium-dosage groups were shortened and the times of crossing target quadrant in space exploration experiment significantly increased(P<0.05, P<0.01)compared with model group. Proteomics results showed that there were 33 differentially expressed proteins between model and Tianqi Yizhi Granules groups. The functions of these proteins and protein network were related to energy metabolism, neurotransmission, cell apoptosis, cytoskeleton and neurotrophy. The Western blot results of malate dehydrogenase and FAS-associated death domain protein were consistent with proteomics results. Conclusion Tianqi Yizhi Granules can improve learning and memory ability of AD rats and the action of mechanism is closely related to the regulation of protein expression of energy metabolism, neurotransmission, apoptosis, cytoskeleton and neurotrophin.
引文
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[11] HAN W N, HOLSCHER C, YUAN L, et al. Liraglutide protects against amyloid-beta protein-induced impairment of spatial learning and memory in rats[J]. Neurobiology of Aging,2013, 34(2):576-588.
[12] MI H, HUANG X, MURUGANUJAN A, et al. PANTHER version 11:expanded annotation data from Gene Ontology and Reactome pathways, and data analysis tool enhancements[J]. Nucleic Acids Research, 2017, 45(D1):D183-D189.
[13] SZKLARCZYK D, MORRIS J H, COOK H, et al. The STRING database in 2017:quality-controlled protein-protein association networks, made broadly accessible[J]. Nucleic Acids Research,2017,45(Dl):D362-D368.
[14] LI Q, WANG J, LI Y, et al. Neuroprotective effects of salidroside administration in a mouse model of Alzheimer's disease[J]. Molecular Medicine Reports,2018, 17(5):7287-7292.
[15] WEI D F, CHEN T, YAN M F, et al. Synthesis, characterization,antioxidant activity and neuroprotective effects of selenium polysaccharide from Radix hedysari[J]. Carbohydrate Polymers,2015,125(2015):161-168.
[16] VAN GIAU V, AN S S A, HULME J P. Mitochondrial therapeutic interventions in Alzheimer's disease[J]. Journal of The Neurological Sciences,2018,395:62-70.
[17] PICKETT E K, ROSE J, MCCRORY C, et al. Region-specific depletion of synaptic mitochondria in the brains of patients with Alzheimer's disease[J]. Acta Neuropathologica,2018, 136(5):747-757.
[2] Alzheimer's Association. 2015 Alzheimer's disease facts and figures[J]. Alzheimer's Dement, 2015, 11(3):332-384.
[3] PRINCE M, WIMO A, GUERCHET M, et al. The global impact of dementia:an analysis of prevalence, incidence, cost and trends[J]. World Alzheimer Report, 2015, 2015:22.
[4] WILSON R S, SEGAWA E, BOYLE P A, et al. The natural history of cognitive decline in Alzheimer's disease[J]. Psychol Aging, 2012,27(4):1008-1017.
[5] VILLEMAGNE V L, BURNHAM S, BOURGEAT P, et al. Amyloid beta deposition, neurodegeneration, and cognitive decline in sporadic Alzheimer's disease:a prospective cohort study[J]. Lancet Neurology, 2013, 12(4):357-367.
[6] MORMINO E C, BETENSKY R A, HEDDEN T, et al. Synergistic effect of beta-amyloid and neurodegeneration on cognitive decline in clinically normal individuals[J]. JAMA Neurology, 2014, 71(11):1379-1385.
[7] TABNER B J, MAYES J, ALLSOP D. Hypothesis:soluble abetaoligomers in association with redox-active metal ions are the optimal generators of reactive oxygen species in Alzheimer's disease[J]. International Journal of Alzheimer's Disease,2010,2011:546380.
[8] MULLANE K, WILLIAMS M. The de-Alzheimerization of age-related dementias:implications for drug targets and approaches to effective therapeutics[J]. Current Opinion in Pharmacology,2018,11:1-14.
[9]武义明,马涛,赵军,等.黄芩苷和牛磺酸对AD模型保护作用的最佳配比研究[J].中国中医药信息杂志,2015, 22(10):54-59.
[10]白卫国,张仰君,徐凯,等.人参提取物治疗阿尔茨海默病药理作用及机制研究进展[J].中国中医药信息杂志,2016, 23(9):126-129.
[11] HAN W N, HOLSCHER C, YUAN L, et al. Liraglutide protects against amyloid-beta protein-induced impairment of spatial learning and memory in rats[J]. Neurobiology of Aging,2013, 34(2):576-588.
[12] MI H, HUANG X, MURUGANUJAN A, et al. PANTHER version 11:expanded annotation data from Gene Ontology and Reactome pathways, and data analysis tool enhancements[J]. Nucleic Acids Research, 2017, 45(D1):D183-D189.
[13] SZKLARCZYK D, MORRIS J H, COOK H, et al. The STRING database in 2017:quality-controlled protein-protein association networks, made broadly accessible[J]. Nucleic Acids Research,2017,45(Dl):D362-D368.
[14] LI Q, WANG J, LI Y, et al. Neuroprotective effects of salidroside administration in a mouse model of Alzheimer's disease[J]. Molecular Medicine Reports,2018, 17(5):7287-7292.
[15] WEI D F, CHEN T, YAN M F, et al. Synthesis, characterization,antioxidant activity and neuroprotective effects of selenium polysaccharide from Radix hedysari[J]. Carbohydrate Polymers,2015,125(2015):161-168.
[16] VAN GIAU V, AN S S A, HULME J P. Mitochondrial therapeutic interventions in Alzheimer's disease[J]. Journal of The Neurological Sciences,2018,395:62-70.
[17] PICKETT E K, ROSE J, MCCRORY C, et al. Region-specific depletion of synaptic mitochondria in the brains of patients with Alzheimer's disease[J]. Acta Neuropathologica,2018, 136(5):747-757.