党参-茯苓配伍调控ERα/PI3K/Akt信号通路改善单侧颈总动脉永久结扎痴呆大鼠的认知障碍OA
Radix codonopsis combined with Poria improves cognitive impairment in rats with unilateral common carotid artery ligation by regulating the ERα/PI3K/Akt signaling pathway
目的 基于UPLC-Q-TOF-MS/MS分析研究党参-茯苓配伍(CRP)治疗痴呆症的作用机制.方法 采用于UPLC-Q-TOF-MS/MS分析单侧颈总动脉永久结扎(UCCA)模型大鼠的 CRP 的入血入脑成分,并对入血和入脑成分进行网络药理学分析.CRP灌胃UCCA 大鼠干预1个月后通过Morris水迷宫评价其学习记忆能力;苏木精-伊红染色观察海马和皮质的病理形态学变化;NeuN染色分析海马和皮层神经元数量变化;采用代谢组学分析 CRP 治疗UCCA大鼠前后脑内代谢物变化差异;酶联免疫吸附测定(ELISA)检测脑内谷氨酸和γ-氨基丁酸(GABA)的含量;免疫组化法考察雌激素受体α(ERα)的蛋白表达;Western blotting分析p-PI3K、PI3K、p-Akt、Akt的蛋白表达.结果 在UCCA大鼠的脑组织和血液中分别鉴定出125种和126种成分,其中,脑组织中发现了89种党参成分和 36 种茯苓成分,血液中发现了85种党参成分和41种茯苓成分.网络药理学分析表明,CRP治疗痴呆主要与调节进入脑组织的成分所影响的PI3K/Akt等信号通路有关.与假手术组比较,模型大鼠的第5天的逃避潜伏期明显延长(P<0.01),平台象限活动时间、海马和皮质的神经元明显损伤且NeuN平均光密度明显下降(P<0.05);脑内谷氨酸和GABA/谷氨酸含量明显升高(P<0.05),ERα、p-PI3K/PI3K、p-Akt/Akt蛋白表达下调(P<0.05).与模型组比较,CRP干预大鼠的第5天的逃避潜伏期明显缩短(P<0.01),穿越平台次数、平台象限活动时间均明显增加(P<0.05),海马和皮质的神经元损伤好转且NeuN平均光密度明显升高(P<0.05);代谢组学分析表明CRP治疗作用主要富集于雌激素信号通路和 GABA 能突触等信号通路.CRP组大鼠脑内的谷氨酸和GABA/谷氨酸含量明显降低(P<0.05),ERα、p-PI3K/PI3K和p-Akt/Akt蛋白表达上调(P<0.05).结论 CRP 能够改善UCCA痴呆大鼠的学习和记忆缺陷,这与其对脑内ERα/PI3K/Akt信号通路的调节有关.
Objective To investigate the mechanism of Radix codonopsis combined with Poria(CRP)for improving cognitive impairment induced by permanent unilateral common carotid artery ligation(UCCA)in rats.Methods UPLC-Q-TOF-MS/MS and network pharmacology analysis were used to analyze the components of CRP in the blood and brain of rats medicated with CRP.Rat models of UCCA were treated with CRP gavage for 1 month,and Morris water maze,HE staining,and NeuN staining were used to assess the therapeutic efficacy.Metabolomics analysis,ELISA,immunohistochemistry,and Western blotting were employed to explore and validate the therapeutic mechanism of CRP.Results A total of 125 chemical constituents were identified in the brain tissue(89 Radix codonopsis components and 36 Poria components)and 126 in blood samples(85 Radix codonopsis components and 41 Poria components)of medicated rats.Network pharmacology analysis revealed that the therapeutic mechanism of CRP on dementia involved primarily the PI3K/Akt signaling pathway,regulated by the brain-targeting constituents of CRP.In UCCA rats,CRP treatment significantly improved their performance in Morris water maze test,alleviated neuronal damage in the hippocampus and cortex,and increased expression level of NeuN.Metabolomics analysis revealed significant enrichment of the estrogen and GABAergic synapse signaling pathways in CRP-treated rats.CRP obviously reduced the brain levels of glutamate and GABA/glutamate in UCCA rats,and downregulated the proteins expressions of ERα,p-PI3K/PI3K and p-Akt/Akt.Conclusion CRP improves learning and memory impairment in UCCA rats possibly by modulating the ERα/PI3K/Akt signaling pathway in the brain.
杨佳瑶;何玉莲;郭延垒;尚芳红;花雷;阳勇;张小梅;魏江平
中药新药创制川渝共建重点实验室//重庆市中药研究院,重庆 400065||国家中医药管理局中药化学三级实验室//重庆市中药研究院 重庆 400065中药新药创制川渝共建重点实验室//重庆市中药研究院,重庆 400065||国家中医药管理局中药化学三级实验室//重庆市中药研究院 重庆 400065中药新药创制川渝共建重点实验室//重庆市中药研究院,重庆 400065中药新药创制川渝共建重点实验室//重庆市中药研究院,重庆 400065||国家中医药管理局中药化学三级实验室//重庆市中药研究院 重庆 400065||重庆中医药学院中药学院,重庆 402760中药新药创制川渝共建重点实验室//重庆市中药研究院,重庆 400065||国家中医药管理局中药化学三级实验室//重庆市中药研究院 重庆 400065中药新药创制川渝共建重点实验室//重庆市中药研究院,重庆 400065||国家中医药管理局中药化学三级实验室//重庆市中药研究院 重庆 400065||川渝共建慢病中药新药泸州市重点实验室,泸州 646000中药新药创制川渝共建重点实验室//重庆市中药研究院,重庆 400065||国家中医药管理局中药化学三级实验室//重庆市中药研究院 重庆 400065||重庆中医药学院中药学院,重庆 402760||川渝共建慢病中药新药泸州市重点实验室,泸州 646000中药新药创制川渝共建重点实验室//重庆市中药研究院,重庆 400065||国家中医药管理局中药化学三级实验室//重庆市中药研究院 重庆 400065||重庆中医药学院中药学院,重庆 402760
党参-茯苓配伍痴呆单侧颈总动脉永久结扎 大鼠γ-氨基丁酸ERα/PI3K/Akt信号通路
compatibility of Radix codonopsis and Poriadementiapermanent unilateral common carotid artery ligationGABAERα/PI3K/Akt signaling pathway
《南方医科大学学报》 2026 (2)
247-258,12
国家自然科学基金青年科学基金(82104427)重庆市巴渝青年岐黄学者支持项目Supported by Natural Science Foundation for the Youth(NSFY)of China(82104427).
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