Chu Hang, Zhang Aihua, Han Ying, Lu Shengwen, Kong Ling, Han Jinwei, Liu Zhidong, Sun Hui, Wang Xijun
National TCM Key Laboratory of Serum Pharmacochemistry, Sino-America Chinmedomics Technology Cooperation Center, Research Center of Chinmedomics (State Administration of TCM), Key Laboratory of Metabolomics, Department of Pharmaceutical Analysis, Heilongjiang University of Chinese Medicine, Heping Road 24, Harbin 150040, China.
National TCM Key Laboratory of Serum Pharmacochemistry, Sino-America Chinmedomics Technology Cooperation Center, Research Center of Chinmedomics (State Administration of TCM), Key Laboratory of Metabolomics, Department of Pharmaceutical Analysis, Heilongjiang University of Chinese Medicine, Heping Road 24, Harbin 150040, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Mar 15;1015-1016:50-61. doi: 10.1016/j.jchromb.2016.02.007. Epub 2016 Feb 8.
Alzheimer's disease (AD) is a multifactorial neurodegenerative disease that influences elderly populations, with no effective method for its treatment so far. To improve its diagnosis and treatment, changes of small molecule metabolite during AD should be elucidated. Kai-Xin-San (KXS) is an herbal formulae that has been widely used to treat mental disorders, especially amnesia and depression in China. Experimental AD was induced in rats by an intraperitoneal injection of d-galactose (d-gal) and administered intragastrically with aluminum chloride (AlCl3) simultaneously for 105 days. Morris water maze task as a behavior test was used for testing the effects of KXS on AD model and pathological changes to the brain were assessed by hematoxylin-eosin staining and immunohistochemistry. The levels of Bcl-2 and ChAT in hippocampus were evaluated by western-blot. Furthermore, metabolite profiling of AD was performed through ultra-performance liquid chromatography/electrospray ionization quadruple time-of- flight-high-definition mass spectrometry (UPLC/ESI-Q-TOF/HDMS) combined with pattern recognition approaches and pathway analysis. d-gal and AlCl3-treated caused a decline in spatial learning and memory, hippocampal histopathological abnormalities and increased Aβ1-40 levels in the brain cortex and hippocampus along with decreased Bcl-2 and ChAT expression in the hippocampus. KXS significantly improved the cognitive impairment induced by d-gal and AlCl3, attenuated hippocampal histopathological abnormalities, reduced Aβ1-40 levels and increased Bcl-2 and ChAT expression in the hippocampus. A total of 48 metabolites were considered as potential biomarkers of AD, and 36 metabolites may correlate with the regulation of KXS treatment on AD. Changes in AD metabolic profiling were close to normal states through regulating multiple perturbed pathways after KXS treatment. This study has revealed the potential biomarkers and metabolic networks of AD, illuminated the biochemistry mechanism of AD and the metabolic pathways influenced by KXS.
阿尔茨海默病（AD）是一种影响老年人群的多因素神经退行性疾病，目前尚无有效的治疗方法。为了改善其诊断和治疗，需要阐明AD病程中小分子代谢物的变化。开心散（KXS）是一种中药方剂，在中国已被广泛用于治疗精神障碍，尤其是失忆和抑郁。通过腹腔注射D-半乳糖（D-gal）诱导大鼠实验性AD，并同时灌胃给予氯化铝（AlCl3），持续105天。采用Morris水迷宫任务作为行为测试来检测KXS对AD模型的影响，并通过苏木精-伊红染色和免疫组织化学评估大脑的病理变化。通过蛋白质免疫印迹法评估海马中Bcl-2和ChAT的水平。此外，通过超高效液相色谱/电喷雾电离四极杆飞行时间高清质谱（UPLC/ESI-Q-TOF/HDMS）结合模式识别方法和通路分析对AD进行代谢物谱分析。D-gal和AlCl3处理导致空间学习和记忆能力下降、海马组织病理学异常，大脑皮层和海马中Aβ1-40水平升高，同时海马中Bcl-2和ChAT表达降低。KXS显著改善了D-gal和AlCl3诱导的认知障碍，减轻了海马组织病理学异常，降低了Aβ1-40水平，并增加了海马中Bcl-2和ChAT的表达。共有48种代谢物被认为是AD的潜在生物标志物，36种代谢物可能与KXS治疗AD的调节作用相关。KXS治疗后通过调节多个受干扰的通路，AD代谢物谱的变化接近正常状态。本研究揭示了AD的潜在生物标志物和代谢网络，阐明了AD的生物化学机制以及KXS影响AD的代谢途径。
