椒红丸及其组方药物对东莨菪碱诱导阿尔茨海默病小鼠的影响

Effects of Jiaohong Pills and Its Prescription on Scopolamine-induced Alzheimer's Disease Mice

目的:通过药效学和代谢组学研究,探讨椒红丸及组方药物对东莨菪碱诱导阿尔茨海默病(AD)模型小鼠认知功能障碍的改善作用及其机制。方法:以东莨菪碱诱导构建AD模型,选用椒红、地黄、椒红丸分别进行干预,采用旷场、水迷宫、物体识别、避暗实验检测小鼠认知功能及胆碱能系统和氧化应激相关生化检测,探究椒红丸及其组方药物药效差异;采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UPLC Q-Exactive Orbitrap MS)对小鼠大脑皮层进行非靶向代谢组学分析。结果:行为学数据显示,与模型组比较,椒红丸高剂量、椒红、地黄组小鼠新物体识别实验中辨别指数均明显增加(P<0.05);椒红、地黄及椒红丸高低剂量组小鼠水迷宫实验中登台率明显增加(P<0.05,P<0.01);椒红、椒红丸高低剂量组小鼠穿台次数明显增加(P<0.05,P<0.01);椒红、椒红丸高剂量组小鼠避暗实验错误次数显著降低(P<0.01),椒红丸及其组方药物组小鼠错误潜伏期均显著升高(P<0.01)。与模型组比较,椒红丸两个剂量组、椒红组小鼠大脑皮层乙酰胆碱转移酶活力明显升高(P<0.05,P<0.01),椒红丸高剂量组乙酰胆碱酯酶活力明显降低(P<0.05),乙酰胆碱水平显著升高(P<0.01),同时椒红丸两个剂量组小鼠血清中丙二醛含量明显降低(P<0.05,P<0.01)。大脑皮层代谢组学研究结果显示,椒红丸组与模型组共鉴定出差异代谢物149个,涉及神经递质代谢、能量代谢、氧化应激与氨基酸代谢功能。结论:椒红丸及其组方药物可以拮抗东莨菪碱造成的认知功能障碍、调节胆碱能系统、降低氧化应激损伤;其对AD治疗作用机制与调控神经递质、能量、氨基酸代谢,改善氧化应激等作用有关。

Objective:To investigate the effects of Jiaohong pills(JHP)and its prescription,Pericarpium Zanthoxyli(PZ)and Rehmanniae Radix(RR)cognitive dysfunction in scopolamine-induced Alzheimer's disease(AD)mice and its mechanism through pharmacodynamic and metabolomics study.Method:The animal model of AD induced by scopolamine was established and treated with PZ,RG and JHP,respectively.The effects of JHP and its formulations were investigated by open field test,water maze test,object recognition test,avoidance test,cholinergic system and oxidative stress related biochemical test.Untargeted metabolomics analysis of cerebral cortex was performed by ultra-performance liquid chromatography-Quadrupole/Orbitrap high resolution mass spectrometry(UPLC Q-Exactive Orbitrap MS).Result:The behavioral data showed that,compared with the model group,the discrimination indexes of the high dose of JHP,PZ and RR groups was significantly increased(P<0.05).The staging rate of Morris water maze test in the PZ,RR,high and low dose groups of JHP was significantly increased(P<0.05,P<0.01),the crossing numbers in the PZ,JHP high and low dose groups were significantly increased(P<0.05,P<0.01);the number of errors in the avoidance test were significantly reduced in the PZ and high-dose JHP groups(P<0.01),and the error latencies were significantly increased in the JHP and its prescription drug groups(P<0.01).Compared with the model group,the activities of acetylcholinesterase in the cerebral cortex of the two doses of JHP group and the PZ group were significantly increased(P<0.05,P<0.01),and the activity of acetylcholinesterase in the highdose JHP group was significantly decreased(P<0.05),and the level of acetylcholine was significantly increased(P<0.01).At the same time,the contents of malondialdehyde in the serum of the two dose groups of JHP decreased significantly(P<0.05,P<0.01).The results of metabolomics study of cerebral cortex showed that 149 differential metabolites were identified between the JHP group and the model group,which were involved in neurotransmitter metabolism,energy metabolism,oxidative stress and amino acid metabolism.Conclusion:JHP and its prescription can antagonize scopolamine-induced cognitive dysfunction,regulate cholinergic system,and reduce oxidative stress damage.The mechanism of its therapeutic effect on AD is related to the regulation of neurotransmitter,energy,amino acid metabolism,and improvement of oxidative stress.