Identification of protein targets for the antidepressant effects of Kai-Xin-San in Chinese medicine using isobaric tags for relative and absolute quantitation

Kai-Xin-San consists of Ginseng Radix, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria at a ratio of 3:3:2:2. Kai-Xin-San has been widely used for the treatment of emotional disorders in China. However, no studies have identified the key proteins implicated in response to Kai-Xin-San treatment. In this study, rat models of chronic mild stress were established using different stress methods over 28 days. After 14 days of stress stimulation, rats received daily intragastric administrations of 600 mg/kg Kai-Xin-San. The sucrose preference test was used to determine depression-like behavior in rats, while isobaric tags were used for relative and absolute quantitation-based proteomics to identify altered proteins following Kai-Xin-San treatment. Kai-Xin-San treatment for 2 weeks noticeably improved depression-like behaviors in rats with chronic mild stress. We identified 33 differentially expressed proteins: 7 were upregulated and 26 were downregulated. Functional analysis showed that these differentially expressed proteins participate in synaptic plasticity, neurodevelopment, and neurogenesis. Our results indicate that Kai-Xin-San has an important role in regulating the key node proteins in the synaptic signaling network, and are helpful to better understand the mechanism of the antidepressive effects of Kai-Xin-San and to provide objective theoretical support for its clinical application. The study was approved by the Ethics Committee for Animal Research from the Chinese PLA General Hospital(approval No. X5-2016-07) on March 5, 2016.

Kai-Xin-San regulates synaptic plasticity through calcium signaling to alleviate symptoms of depression-like behavioral disorders in mice

Background:Kai-Xin-San,a classical Chinese medicine prescription,has been widely applied in the clinical therapy for depression,but its pharmacological mechanism remains to be further explored.Based on network pharmacology,molecular docking and animal experiments,the research is performed to exploit pharmacological mechanism of Kai-Xin-San for treating depression.Methods:Obtain chemical components and potential targets of Kai-Xin-San through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,Encyclopedia of Traditional Chinese Medicine and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine databases,and then screen the active ingredients of each herb in accordance with absorption,distribution,metabolism,and excretion.The GenCards,Online Mendelian Inheritance in Man,Therapeutic Target database and DrugBank databases were used to obtain the major targets of depression,and the STRING platform was used to construct the protein-protein interaction network and explore the potential protein functional modules in the network.The targets were subjected to Gene Ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis by STRING database and Metascape database.The interaction network of“Kai-Xin-San active components-depression-targets-pathways”was constructed by Cytoscape,and molecular docking verification was performed by Auto Dock tools.Finally,animal experiments were carried out for further verification.The chronic restraint stress depression model was established and mice were randomly divided into 4 groups:control group,chronic restraint stress group,fluoxetine group and Kai-Xin-San group.Behavioral tests were used to evaluate the depressive phenotype of mice.The expression of CaMKII-,synaptophysin,poststroke depression-95,and CACNA1C were all detected using a western blot.Results:Network analysis shows that Kai-Xin-San may mainly regulate calcium signaling pathway to exert antidepressant effects.A majority of the targets and components have good binding activity,according to the molecular docking studies.In the current study,behavioral tests showed that Kai-Xin-San could effectively alleviate depression-like behaviors in mice compared with the chronic restraint stress group,which effect was comparable to fluoxetine.Meanwhile,compared with the chronic restraint stress group,protein levels of CACNA1C,CaMKII-α,synaptophysin and poststroke depression-95 were significantly increased(P<0.05).Conclusion:The research initially identifies the multi-component,multi-target,and multi-path mechanism of Kai-Xin-San in the treatment of depression.Kai-Xin-San may improve synaptic plasticity through calcium signaling pathway to exert antidepressant effects.

Characterization of chemical constituents and in vivo metabolites of Kai-Xin-San prescription by HPLC/DAD/ESI-MS^n

This work aims to elucidate the chemical constituents of Kai-Xin-San（KXS） and its metabolites in rat plasma.KXS extracts were separated on an Agilent HPLC SB-C 18 column,analyzed by ion-trap tandem mass spectrometry and high-accuracy qTOF mass spectrometry in negative ion mode.A total of 39 compounds,including 11 ginsenosides,14 Polygala saponins,5 sucrose esters,8 oligosaccharide esters and 1 xanthone were characterized from KXS.Fifteen of them were confirmed by reference standards.No constituents were detected from Poria or Acori Tatarinowii Rhizoma.After oral administration of KXS（7 g/kg）,10 ginsenosides and 18 Polygala compounds were detected in rat plasma.This study indicates that ginseng saponins,Polygala saponins and saccharide esters could be the major effective components of KXS prescription.

开心散联合氟西汀改善慢性压力应激小鼠抑郁样行为的效应评价研究

目的考察中药复方开心散联合用药氟西汀改善慢性压力应激小鼠抑郁样行为的效用与作用机制。方法采用慢性压力应激构建抑郁小鼠模型,通过糖水偏嗜实验评价开心散与氟西汀联合用药改善模型小鼠抑郁样行为的效用。采用ELISA法测定受试动物血清、海马组织和下丘脑组织中皮质醇(Cortisol)、促肾上腺皮质激素(ACTH)和促肾上腺皮质激素释放因子(CRF)的表达。采用HE染色、尼氏染色和TUNEL染色法,观察模型小鼠海马神经元损伤情况。采用Western blot法考察小鼠海马组织中Caspase-3和Caspase-9等凋亡信号通路关键蛋白的表达情况。结果开心散与氟西汀联用能显著上调模型小鼠糖水偏嗜率;同时,联合用药可抑制脑内和血清中压力应激因子的上调,减少小鼠海马区因抑郁导致的细胞凋亡,调控海马区的凋亡信号通路。结论开心散联合氟西汀可有效改善抑郁模型动物的抑郁样行为,减少氟西汀临床用量。两者联用后抑制压力应激轴激活可能是联合用药抗抑郁的作用环节。

中药复方开心散调控慢性压力应激小鼠海马神经新生抗抑郁作用机制研究

目的评价开心散调控慢性不可预知压力应激模型小鼠海马神经新生抗抑郁效用,探讨作用机制。方法采用慢性不可预知压力应激构建抑郁小鼠模型,给以开心散水提取物。通过糖水偏嗜率检测、强迫游泳实验、悬尾实验等评价模型小鼠抑郁样行为。采用免疫荧光法检测模型小鼠海马区神经新生情况。采用蛋白免疫印迹法检测小鼠海马神经干细胞标志物巢蛋白以及Wnt/β-catenin信号通路相关蛋白的表达。采用ELISA法检测相应组织中压力应激因子如促肾上腺皮质激素释放因子、促肾上腺皮质激素、皮质醇与神经营养因子如脑源性神经营养因子、神经生长因子的表达。采用小鼠原代神经干细胞,采用免疫荧光法检测开心散对原代神经干细胞增殖情况,采用蛋白免疫印迹法检测巢蛋白以及Wnt/β-catenin信号通路相关蛋白的表达。结果开心散水提取物显著增加抑郁模型小鼠糖水偏嗜率,降低强迫游泳与悬尾不动时间;同时促进海马组织神经元新生,降低压力应激因子表达,提高神经营养因子表达,上调巢蛋白表达并调控Wnt/β-catenin信号通路。开心散提取物还可促进小鼠原代神经干细胞增殖及蛋白表达,并促进β-catenin入核,调控Wnt/β-catenin信号通路。结论开心散具有改善小鼠抑郁样行为效用,促进海马神经新生以及调控Wnt/β-catenin通路可能是其抗抑郁的重要作用环节与机制。

开心散联合氟西汀对慢性压力应激抑郁小鼠肠道菌群与机体致炎物质表达的影响

目的评价开心散联合氟西汀对慢性压力应激(CUMS)抑郁小鼠肠道菌群与机体致炎物质表达的影响,探索其调控肠-脑轴抗抑郁作用机制。方法构建小鼠CUMS抑郁模型,通过检测糖水偏嗜率、悬尾不动时间与强迫游泳不动时间评价开心散联合氟西汀改善小鼠抑郁行为的效果。采用ELISA法测定受试动物大脑皮层、血清和小肠组织中致炎物质白介素6(IL-6)、白介素1β(IL-1β)、肿瘤坏死因子-α(TNF-α)与脂多糖(LPS)的表达。采用16S rRNA基因测序法对小鼠粪便中肠道菌群组成进行分析。采用Western blot法考察小鼠小肠组织中ZO-1、Occludin和Claudin 5等肠道屏障蛋白的表达情况。结果开心散联合氟西汀显著上调模型小鼠糖水偏嗜率(P<0.01),缩短悬尾不动时间与强迫游泳不动时间(P<0.05,P<0.01)。开心散与氟西汀中剂量联用的抗抑郁效用与单用氟西汀高剂量的趋势相似。联合用药还可显著抑制CUMS模型动物大脑皮层、血清和小肠致炎物质的上调(P<0.05,P<0.01)。肠道菌群分析显示,联合用药可改善模型动物肠道革兰氏阳性菌与阴性菌的比例,其对毛螺菌、双歧杆菌、瘤胃球菌、布劳特氏菌、优杆菌、丁酸肠杆菌、丹毒丝菌、另枝菌、脱硫弧菌、红椿菌-UCG-002等肠道菌相对丰度的改善与其缓解抑郁样行为与皮层致炎物质表达下调显著相关(P<0.05,P<0.01)。另外,联合用药还可显著上调肠道屏障蛋白表达(P<0.05,P<0.01)。结论开心散联合氟西汀可缓解CUMS模型动物的抑郁样行为并减少氟西汀单用的用量。两者连用调控肠道菌群组成,抑制肠道致炎物质表达,上调肠道屏障蛋白表达,从而减少血清与中枢的致炎物质的表达,可能是其调控肠-脑轴抗抑郁的作用环节。

开心散对Aβ_(25-35)诱导损伤SH-N-K神经细胞的保护作用及机制

目的观察开心散对神经毒性物质β-淀粉样肽(Aβ25-35)诱导损伤的SK-N-SH细胞的保护作用。方法用Aβ25-35处理人神经母细胞瘤细胞(SK-N-SH细胞),模拟阿尔茨海默病(AD)病人脑内神经元病理损伤,并以含开心散(KXS)药物血清拮抗其作用。以MTT法测定细胞存活率,酶反应法检测细胞内超氧化物歧化酶SOD水平变化,流式细胞技术检测细胞凋亡率。逆转录聚合酶链式反应(RT-PCR)检测淀粉样肽蛋白前体(APP)基因、Aβ降解酶NEP基因的表达情况,Westernblots法检测APP蛋白表达。结果试验显示暴露Aβ25-35后培养SK-N-SH细胞存活率明显下降,凋亡细胞比例增加。而含开心散药物血清处理能显著提高Aβ25-35损伤细胞的存活率,减少细胞凋亡率,同时提高细胞内SOD的活力(P<0.01)。RT-PCR结果显示开心散可降低Aβ代谢相关的APP基因的表达并提高NEP基因的表达,APP蛋白表达显著减少。结论AD病理相关的Aβ25-35能造成神经元细胞损伤死亡,开心散对毒性Aβ25-35诱导的神经细胞损伤具有一定保护作用,其作用机制可能与开心散能增加抗氧化酶SOD活力,同时下调内源性APP基因表达,上调NEP基因表达,减少内源Aβ产生,从而抑制细胞死亡有关。

开心散有效部位对小鼠学习记忆功能的影响

用药理研究指导植化分离 ,寻找中药复方开心散有效部位 (KXS) ,探讨其益智作用及机理。采用氢溴酸东莨菪碱造模 ,用跳台法和水迷宫测试小鼠的学习记忆能力 ,测定小鼠大脑组织乙酰胆碱酯酶 (AchE)和超氧化物歧化酶 (SOD)活性以探讨其作用机理。跳台法测试表明模型组正确区时间 (Ts)缩短 ,KXS能延长Ts ,水迷宫实验结果显示 ,模型小鼠游泳时间明显延长 ,KXS能缩短游泳时间。KXS能逆转模型小鼠AchE活性的升高和SOD活性的降低。实验结果提示KXS对模型小鼠的学习记忆能力有一定的改善作用。

开心散对拟痴呆小鼠记忆功能的影响

目的 探讨 Ａ ｌｚｈｅｉｍ ｅｒ 病（ Ａ Ｄ）与铝的关系以及中药对 Ａ Ｄ 的治疗作用。方法 实验采用氯化铝（ Ａ ｌ Ｃｌ３ ）制作小鼠拟痴呆模型，研究中药开心散对小鼠学习记忆和超氧化物歧化酶与脂质过氧化物（ Ｓ Ｏ Ｄ／ Ｍ Ｄ Ａ ）活性及脑组织蛋白含量的影响。结果 Ａ ｌ Ｃｌ３ 中毒小鼠记忆能力明显降低，表现为在电、水迷宫中准确次数减少，准确率降低。血清、肝、心、脑组织 Ｓ Ｏ Ｄ 活性下降；心、肝组织 Ｍ Ｄ Ａ 含量升高；脑组织蛋白含量降低。模型鼠服用开心散后，记忆能力明显提高， Ｓ Ｏ Ｄ 活性升高， Ｍ Ｄ Ａ 降低，脑组织蛋白含量升高。结论 开心散具有明显益智健脑、防衰老、增强记忆功能。

基于中医方证代谢组学的开心散干预老年痴呆症大鼠的效应物质动态分析

目的:本研究在开心散干预氯化铝联合D-半乳糖诱导老年痴呆症大鼠模型有效性的基础上,揭示开心散对老年痴呆症(AD)不同阶段干预作用的代谢物靶点,构建开心散体内成分与内源性生物标记物的网络关系,分析开心散对AD发展过程干预作用的效应物质的动态变化。方法:利用行为学指标评价模型复制进程以及开心散的干预作用;运用代谢组学技术和方法分析开心散对AD大鼠模型不同阶段干预作用的靶点;应用UPLC-MS技术和MSE质谱数据采集模式,分别对开心散体外化学成分和口服给药后的血中移行成分进行表征及鉴定;利用PCMS手段,构建复方药效物质基础与内源性生物标记物的数理模型,发掘开心散防治老年痴呆症不同阶段的效应物质。结果:给予AD大鼠开心散溶液治疗后,模型大鼠的认知功能得到改善,其病理学进程得以延缓。代谢组学研究表明,开心散对AD模型不同阶段标记物的异常表达起到恢复作用,体现了开心散干预老年痴呆症多靶点、多途径的特点。结论:基于中医方证代谢组学方法为开心散减缓AD进程研究提供了新的思路。

老年大鼠神经、内分泌、血流变学改变及开心散的影响

目的 研究老年大鼠神经、内分泌、血液流变学的改变 ,探讨衰老的发病机理及中药开心散益智抗衰老的作用机理。 方法 用 2 1月龄大鼠作为衰老模型 ,并用中药开心散进行治疗 ;6月龄大鼠为青年对照组。测定血浆皮质醇、血液流变学及脑内单胺类神经递质的含量变化。 结果 老年大鼠血浆皮质醇含量较高 ;全血和血浆粘度明显升高 ;脑内多巴胺 (DA)含量明显降低 ,去甲肾上腺素 (NE)、5 羟吲哚乙酸 ( 5 HIAA)亦有下降 ,5 羟色胺 ( 5 HT)升高。应用开心散后老年大鼠皮质醇含量下降 ;血液粘度改善 ;脑组织DA、NE、5 HIAA含量升高 ,5 HT含量降低。 结论 2 1月龄大鼠可作为衰老动物模型 ;衰老的形成与神经、内分泌、血液流变学改变等综合因素有关 ;中药开心散具有延缓衰老的作用 ,其抗衰老作用可能与改善大鼠下丘脑 垂体 肾上腺轴功能衰退和血液流变性有关。

开心散配伍比例对慢性压力应激抑郁小鼠皮层与海马神经营养因子系统调控的影响

目的基于慢性压力应激抑郁小鼠模型,考察不同配伍比例的开心散对皮层与海马神经营养因子调控的影响,探讨开心散治疗抑郁症的作用机制。方法采用一系列慢性应激压力构建小鼠抑郁症模型,给予不同配伍比例的开心散,应用ELISA法分别检测小鼠大脑皮层与海马中神经生长因子NGF与脑源性神经营养因子BDNF含量,利用qPCR与Western blot技术检测皮层与海马神经营养因子受体TrkA、TrkB、TrkC的表达。结果不同配伍比例的开心散均能提升抑郁小鼠的糖水饮用量,延长强迫游泳时间,表现出显著的抗抑郁作用,其中以D-652配伍比例效用最佳。同时提升NGF、BDNF含量与相关受体的表达。结论开心散调控神经营养因子系统是其抗抑郁的重要作用机制。

中药复方开心散调控神经营养因子抗抑郁物质基础与作用机制研究

目的:观察开心散调控神经营养因子抗抑郁的物质基础与作用机制。方法:制备开心散大孔树脂乙醇洗脱部位,采用悬尾和强迫游泳评价抗抑郁效果。采用ELISA法测定海马中神经生长因子(NGF)与脑源性神经营养因子(BDNF)的含量。利用大鼠星型胶质瘤C6细胞系和大鼠肾上腺嗜铬瘤PC12细胞系,评价不同乙醇洗脱部位促进NGF与BDNF表达和促进PC12细胞分化的作用。结果:开心散各效用部位均能显著缩短小鼠悬尾与强迫游泳不动时间,其中70%乙醇洗脱部位抗抑郁效用最强,该部位还能通过c AMP信号通路上调C6细胞中NGF与BDNF的表达。10%乙醇洗脱部位促PC12细胞分化的能力最强。人参皂苷是促进神经营养因子表达的主要效用成分。结论:开心散调控神经营养因子是其抗抑郁效用的重要作用机制。

中药复方开心散调控慢性压力应激小鼠海马炎性细胞因子水平抗抑郁作用机制研究

目的探究中药复方开心散调控中枢神经炎症系统抗抑郁的作用机制。方法制备开心散不同配伍比例提取物,采用小鼠慢性应激压力(CUMS)模型,通过糖水偏嗜实验、悬尾实验和强迫游泳实验评价开心散不同配伍比例的抗抑郁功效。采用ELISA法测定受试动物海马组织促炎物质如脂多糖(LPS)、IL-1β、IL-6、TNF-α的含量。采用免疫组织化学染色法观察受试动物海马区小胶质细胞形态。进而采用LPS促进小鼠小胶质永生化细胞(BV2)炎性因子表达细胞模型,采用ELISA法评价开心散抑制炎症因子表达的作用,采用Western blotting法考察NF-κB信号通路的调控情况。结果开心散不同配伍比例能显著上调模型小鼠糖水偏嗜率,缩短小鼠悬尾和强迫游泳不动时间,以D-652效用最强。该配伍比例通过抑制NF-κB的入核和抑制NF-κB磷酸化等环节,下调BV2细胞中炎症因子IL-1β、IL-6、TNF-α的表达水平。结论对中枢神经炎症的调控是中药复方开心散抗抑郁的重要作用环节。

运用网络药理学探讨开心散治疗抑郁症的作用机制

目的本研究旨在运用网络药理学方法,预测开心散活性成分的抗抑郁药理作用机制。方法通过TCMSP、ETCM获取开心散的成分和靶点,运用Genecard、OMIM筛选抑郁症相关的蛋白,通过STRING构建蛋白相互作用网络,使用Cytoscape进行靶点的拓扑分析、MCODE聚类分析,使用R语言进行GO和KEGG通路富集分析,最后使用MOE软件进行分子对接验证。结果网络分析表明,开心散中25个活性成分与其发挥抗抑郁作用密切相关,该机制涉及到51个靶点(包括ACHE、HTR3A、DRD2等)以及24条通路;分子对接结果显示化合物aposcopolamine、perlolyrine、hederagenin与PPARG、ACHE、SLC6A4、DRD2靶点均具有较好的结合能力。结论开心散可能主要通过神经活性受体配体相互作用、羟色胺突触、cAMP通路等相关靶标起到抗抑郁的功效,该研究为一步揭示开心散治疗抑郁症的潜在活性成分及其可能的作用机制提供了思路。

开心散与生脉散抗阿尔茨海默症“同病异治”作用机制网络药理学分析与中枢神经免疫调控效用验证

目的基于网络药理学方法研究开心散(KXS)与生脉散(SMS)“同病异治”抗阿尔茨海默症作用机制,并通过整体动物与离体细胞实验进行验证。方法通过数据库搜集筛选开心散与生脉散复方所含药味化学成分并预测其潜在靶点,构建复方-中药-成分-靶点网络。筛选复方共有与优势靶点,构建蛋白互作网络并进行拓扑分析,进行KEGG通路和GO功能富集分析。基于网络药理学分析获得的开心散与生脉散共性与个性的生物学GO分析结果,分别建立小鼠海马Aβ注射拟阿尔茨海默症痴呆小鼠模型与小鼠小胶质BV2细胞炎性因子损伤模型,检测相应指标,采用行为学结合生化指标分析验证网络药理学分析结果。结果筛选出开心散与生脉散复方共有成分有109个,共有靶点490个,抗AD潜在共性调控靶点375个。开心散抗AD潜在优势调控靶点92个,生脉散抗AD潜在优势调控靶点90个。淀粉样蛋白代谢与转运、cAMP与AMPK信号通路、突触功能调控等为开心散与生脉散抗AD潜在共性调控信号通路与生物学事件。NF-κB等为开心散抗AD优势调控信号通路与生物学事件;趋化因子、胆碱能突触等为生脉散抗AD优势调控信号通路与生物学事件。验证实验结果显示开心散与生脉散能够有效改善Aβ海马区注射小鼠认知功能障碍,降低小鼠海马和小胶质细胞的炎性因子表达。结论开心散与生脉散能通过共有及各自优势靶点发挥抗AD作用。

开心散所含药对干预应激抑郁大鼠行为、中枢神经递质及5-HT1A表达的研究思路与方法

采用药对分组方法,通过对抑郁大鼠的一般状态、行为、海马组织NE、5-HT和5-HT1A表达的分析,采用药对分组方法研究开心散及其所包含的药对抗抑郁作用的差异,揭示开心散方中蕴涵的配伍规律,为临床抗抑郁提供基础数据。

Kai Xin San ameliorates scopolamine-induced cognitive dysfunction

Kai Xin San(KXS, containing ginseng, hoelen, polygala, and acorus), a traditional Chinese herbal compound, has been found to regulate cognitive dysfunction; however, its mechanism of action is still unclear. In this study, 72 specific-pathogen-free male Kunming mice aged 8 weeks were randomly divided into a vehicle control group, scopolamine group, low-dose KXS group, moderate-dose KXS group, high-dose KXS group, and positive control group. Except for the vehicle control group and scopolamine groups(which received physiological saline), the doses of KXS(0.7, 1.4 and 2.8 g/kg per day) and donepezil(3 mg/kg per day) were gastrointestinally administered once daily for 2 weeks. On day 8 after intragastric treatment, the behavioral tests were carried out. Scopolamine group and intervention groups received scopolamine 3 mg/kg per day through intraperitoneal injection. The effects of KXS on spatial learning and memory, pathological changes of brain tissue, expression of apoptosis factors, oxidative stress injury factors, synapse-associated protein, and cholinergic neurotransmitter were measured. The results confirmed the following.(1) KXS shortened the escape latency and increased residence time in the target quadrant and the number of platform crossings in the Morris water maze.(2) KXS increased the percentage of alternations between the labyrinth arms in the mice of KXS groups in the Y-maze.(3) Nissl and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining revealed that KXS promoted the production of Nissl bodies and inhibited the formation of apoptotic bodies.(4) Western blot assay showed that KXS up-regulated the expression of anti-apoptotic protein Bcl-2 and inhibited the expression of pro-apoptotic protein Bax. KXS up-regulated the expression of postsynaptic density 95, synaptophysin, and brain-derived neurotrophic factor in the cerebral cortex and hippocampus.(5) KXS increased the level and activity of choline acetyltransferase, acetylcholine, superoxide dismutase, and glutathione peroxidase, and reduced the level and activity of acetyl cholinesterase, reactive oxygen species, and malondialdehyde through acting on the cholinergic system and reducing oxidative stress damage. These results indicate that KXS plays a neuroprotective role and improves cognitive function through reducing apoptosis and oxidative stress, and regulating synapse-associated protein and cholinergic neurotransmitters.

Neuroprotective mechanism of Kai Xin San: upregulation of hippocampal insulin-degrading enzyme protein expression and acceleration of amyloid-beta degradation

Kai Xin San is a Chinese herbal formula composed of Radix Ginseng, Poria, Radix Polygalae and Acorus Tatarinowii Rhizome. It has been used in China for many years for treating amnesia. Kai Xin San ameliorates amyloid-β （Aβ） induced cognitive dysfunction and is neuroprotective in vivo, but its precise mechanism remains unclear. Expression of insulin-degrading enzyme （IDE）, which degrades Aβ, is strongly correlated with cognitive function. Here, we injected rats with exogenous Aβ42 （200 μM, 5 μL） into the hippocampus and subsequently administered Kai Xin San （0.54 or 1.08 g/kg/d） intragastrically for 21 consecutive days. Hematoxylin eosin and Nissl staining revealed that Kai Xin San protected neurons against Aβ-induced damage. Furthermore, enzyme linked immunosorbent assay, western blot and polymerase chain reaction results showed that Kai Xin San decreased Aβ42 protein levels and increased expression of IDE protein, but not mRNA, in the hippocampus. Our findings reveal that Kai Xin San facilitates hippocampal Aβ degradation and increases IDE expression, which leads, at least in part, to the alleviation of hippocampal neuron injury in rats.

基于VOSviewer和CiteSpace的开心散研究现状及热点可视化分析

目的探讨开心散的研究现状及热点。方法检索中国知识资源总库(CNKI)、中国学术期刊数据库(万方数据)、中文科技期刊数据库(重庆维普)、中国生物医学文献数据库(CBM)、PubMed及Web of Science建库至2023年1月10日发表的开心散研究文献。采用CiteSpace 6.2.R2和VOSviewer 1.6.16软件对纳入文献的文献类型、来源期刊、发文量、作者、机构、关键词等数据进行可视化分析。结果纳入文献235篇,以中文期刊论文为主,涉及来源期刊87种,其中《中国中药杂志》和J.Ethnopharmacol发文较多。开心散年发文量整体呈上升趋势,涉及505位作者,形成了以刘屏、姜艳艳等为核心的研究团队;纳入文献作者来自99家研究机构,机构间的合作以地理区域相同和相近单位、中医药大学及其附属医院为主;关键词共现聚类网络、关键词共现时间网络和关键词突现分析结果表明,开心散主要活性成分组成(人参皂苷类、茯苓酸、细辛醚类、酮类和寡糖酯类)、检测方法(HPLC及液质联用)、开心散药理作用(抗老年痴呆、抗抑郁)、作用机制及临床应用是当前研究热点和发展动向。结论开心散的研究多集中于AD、抑郁症等疾病作用机制和临床研究,其中开心散主要活性成分研究是近年热点内容,而药理作用机制及临床应用等发展趋势较好,基于组方活性成分与功效相关性可能成为开心散研究新的热点方向。