山茱萸环烯醚萜苷在体外对胆碱酯酶活性的影响

该研究的目的是观察山茱萸环烯醚萜苷(CIG)在体外试验中对胆碱酯酶的作用,以探讨该药在治疗阿尔茨海默病(AD)中的作用机制.使用人红细胞、大鼠全脑匀浆和人血浆作为酶源,采用生物化学方法测定乙酰胆碱酯酶(AChE)和丁酰胆碱酯酶(BuChE)的活性,检测CIG对胆碱酯酶的影响.结果显示,CIG对人红细胞及大鼠全脑匀浆AChE活性具有抑制作用,50％抑制率(IC50)分别为1.6,3.3g·L-1,并且CIG对AChE的抑制是可逆的.CIG对人血浆BuChE也具有抑制作用,IC502.9 g·L-1.实验结果表明,CIG在体外试验中对AChE和BuChE具有一定的抑制作用,这可能为GIG治疗AD的作用机制之一.     

脑益康对阿尔茨海默病大鼠中枢胆碱能系统酶的影响

目的 观察中药复方脑益康对阿尔茨海默病(Alzheimer's disease,AD)大鼠中枢胆碱能系统的保护作用.方法 采用脑立体定位仪自双侧基底前脑Meynert核(nucleus basalis of Meynert,NBM)注射鹅膏蕈氨酸(Ibotenic acid,IBO)建立AD大鼠模型,分别给予不同剂量脑益康对AD大鼠进行治疗,测定不同组实验大鼠脑组织乙酰胆碱酯酶(acetylcholinest-erase,AChE)、丁酰胆碱酯酶(butyrocholinesterase,BuChE)活性及胆碱乙酰转移酶(choline acetyltransferase,ChAT)在基底前脑的表达.结果 脑益康能抑制脑组织AChE和BuChE的活性;改善基底前脑ChAT阳性神经细胞形态并增加其数量.结论 脑益康通过促进ChAT表达,增加乙酰胆碱(acetylcholine,ACh)的合成;通过降低脑组织AChE和BuChE的活性,抑制ACh分解;从而增加脑内ACh含量,改善AD大鼠的学习、记忆障碍.     

苦参制剂对体外血清胆碱酯酶和转氨酶活性的影响

目的探讨临床常用苦参制剂（肝炎灵、苦参碱和苦参素）对血清胆碱酯酶和转氨酶活性的影响。方法采用体外药物与血清共育方式,分别于药物与血清共育10min和24h后,用全自动生化分析仪检测血清胆碱酯酶和转氨酶的活性。结果只有苦参碱对血清胆碱酯酶和转氨酶活性有直接抑制作用,苦参素和肝炎灵对血清胆碱酯酶和转氨酶活性均无直接抑制作用。结论苦参碱能直接抑制血清胆碱酯酶和转氨酶活性;肝炎灵和苦参素对体外血清胆碱酯酶和转氨酶活性无直接抑制作用,其体内用药致血清胆碱酯酶活性下降的原因是其在体内经过代谢转化成为苦参碱或其他具有强力抑制胆碱酯酶活性的代谢产物或转化产物。     

河南产连翘叶抑制α-糖苷酶活性成分研究

目的:寻找连翘叶中α-葡萄糖苷酶抑制活性的成分.方法:利用体外α-葡萄糖苷酶抑制模型活性追踪,采用各种色谱法分离,运用多种波谱技术鉴定结构,并对活性较强的单体化合物进行酶抑制动力学研究.结果:连翘叶的乙酸乙酯部分具有较高的活性,从中分离出5个活性化合物,对α-葡萄糖苷酶抑制活性高于阳性对照阿卡波糖.酶抑制动力学反应结果表明,化合物1,4和1与2混合物(2:1)对α-葡萄糖苷酶的抑制类型为非竞争性抑制剂.结论:化合物1-3为首次发现对α-糖苷酶的抑制活性,3为首次从连翘属中分离得到.     

新型体外胆碱酯酶抑制剂筛选体系的构建

〔摘 要〕 目的： 构建新型简便可靠的体外胆碱酯酶抑制剂筛选体系。方法： 从 SD 大鼠大脑和血清中提取乙酰胆碱酯酶 （AChE）和丁酰胆碱酯酶（BuChE），应用 Ellman 比色法构建体外 AChE 和 BuChE 抑制剂筛选实验体系。结果： 选择 10 倍 稀释的雄性大鼠脑匀浆液〔蛋白浓度（1.69 ± 0.10）mg·mL-1〕和 5 倍稀释的血清作为 AChE/BuChE 酶源，使用 AChE/BuChE 抑制剂他克林作为参考。他克林对 AChE/BuChE 的半数抑制浓度（IC50）分别为 262 nmol·L-1 和 168 nmol·L-1，体外 胆碱酯酶抑制剂筛选体系构建成功。在等量蛋白浓度情况下，雌性大鼠脑匀浆液中 AChE 酶活性为雄性大鼠的 1.84 倍。 结论： 本研究从大鼠大脑和血清里分别提取 AChE 和 BuChE 粗酶，构建了一种新型的简单易行且稳定可靠的 AChE/BuChE 抑制剂筛选平台。     

苦参素体外对血清ALT AST和CHE活性影响的实验研究

目的:采用体外药物与血清共育方式来探讨苦参素对血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)和胆碱酯酶(CHE)活性的影响。方法:采用肝炎灵和苦参素体外与血清共育来观察其对ALT、AST和CHE活性的影响。结果:体外血清共育实验表明,含有苦参碱与苦参素的肝炎灵原液对血清胆碱酯酶活性有直接抑制作用,而苦参素体外对血清胆碱酯酶活性无直接抑制作用。低浓度的肝炎灵和苦参素体外对血清ALT、AST的活性反而有促进作用。结论:苦参素体外对血清胆碱酯酶活性无直接抑制作用,而苦参素体内用药致血清胆碱酯酶明显下降原因是其在人体内可经过代谢转化成为具有强力抑制胆碱酯酶活性的代谢产物或转化产物。     

表小檗碱体外大鼠肝微粒体孵育代谢产物鉴定及对CYP2D6酶活性的抑制作用

表小檗碱是黄连中一种主要的异喹啉类生物碱,具有多种重要的药理活性。该实验利用体外大鼠肝微粒体孵育体系,采用LC-MS/MS分析鉴定表小檗碱体外大鼠肝微粒体(RLM)共孵育后的Ⅰ相和Ⅱ相代谢产物,采用cocktail探针药物法,通过同时测定美托洛尔、氨苯砜、非那西丁、氯唑沙宗、甲苯磺丁脲的含量变化,评价不同浓度表小檗碱对大鼠肝微粒体CYP2D6,CYP3A4,CYP1A2,CYP2E1,CYP2C9亚型活性的影响。结果表明,表小檗碱在大鼠肝脏可发生Ⅰ相和Ⅱ相代谢,从表小檗碱的大鼠体外肝微粒体温孵体系中鉴定出2个Ⅰ相代谢产物和3个Ⅱ相代谢产物;表小檗碱对肝脏CYP2D6酶呈显著抑制作用,半数抑制浓度IC50为35.22μmol·L-1,而对CYP3A4,CYP1A2,CYP2E1,CYP2C9酶的活性没有明显的影响,提示表小檗碱可能存在基于CYP2D6酶的药物相互作用。该研究可为合理开发利用表小檗碱提供实验依据。     

不同浓度白芷美白液对人体酪氨酸酶活性抑制情况分析

目的:对比不同浓度白芷美白液对人体酪氨酸酶活性的抑制情况,探究白芷美白液的作用机制和效果。方法:选用不同浓度(25%、40%、50%、100%)白芷美白液作为研究对象,考察其体外对酪氨酸酶的抑制作用及对细胞内酪氨酸酶活性的抑制作用。结果:不同浓度白芷美白液均可抑制酪氨酸酶活性,其浓度范围在25%~50%时,对酪氨酸酶的抑制作用具有明显量效关系,当白芷美白液的浓度超过50%时,量效关系不明显。白芷美白液对酪氨酸酶的抑制作用属于非竞争性可逆抑制。结论:在一定范围内,白芷美白液浓度增高可增强对酪氨酸酶活性的抑制作用,该作用属于非竞争性可逆抑制。     

清开灵注射剂等5种中药注射剂对大鼠肝微粒体 CYP3A的体外抑制作用

目的:考察清开灵注射剂、金纳多注射剂、疏血通注射剂、参麦注射剂、康艾注射剂这5种临床常用的中药注射剂对大鼠肝微粒体CYP3A的体外抑制作用,预测发生药物相互作用的可能性,以确保这些中药注射剂临床应用的安全性与有效性.方法:采用SD大鼠肝微粒体体外孵育法,在孵育体系中加入底物睾酮和不同体积的清开灵注射剂等5种中药注射剂,用高效液相色谱法测定睾酮的羟化代谢产物6β-羟基睾酮的生成量反映CYP3A的活性,酮康唑用作阳性对照药.结果:在体外孵育体系中,10%的清开灵注射剂大约能抑制93.0%的6β-羟基睾酮生成,抑制效果明显高于其他4种相同浓度的中药注射剂.根据其抑制动力学曲线,计算出清开灵注射剂的IC90和Ki值分别为1.O%和0.7%.结论:在体外系统中,金纳多注射剂和疏血通注射剂对大鼠肝微粒体CYP3A无抑制作用,参麦注射剂和康艾注射剂对CYP3A显示出弱的抑制作用,清开灵注射剂对CYP3A有明显的抑制作用.提示当清开灵注射剂与经CYP3A代谢的药物联合用药时可能会发生药物相互作用,临床联合用药须谨慎.     

胡芦巴生品、酒制品抑制α-葡萄糖苷酶和α-淀粉酶活性的有效部位筛选及其酶动力学分析

目的:比较胡芦巴生品和酒制品对α-葡萄糖苷酶,α-淀粉酶抑制活性作用的差别,筛选抑制活性的有效部位,并研究其酶反应动力学。方法:利用萃取法获得胡芦巴生品和酒制品80%甲醇提取物的石油醚层、乙酸乙酯层、正丁醇层、水层4个不同极性部位,采用体外α-葡萄糖苷酶抑制模型和α-淀粉酶抑制模型,测定生胡芦巴和酒胡芦巴4个不同极性部位对α-葡萄糖苷酶,α-淀粉酶活性的抑制作用。通过酶促动力学与Lineweaver-Burk曲线推断各有效部位对α-葡萄糖苷酶及α-淀粉酶抑制类型。结果:生胡芦巴和酒胡芦巴只有水层对α-葡萄糖苷酶有抑制作用,酒制品对α-葡萄糖苷酶的抑制作用优于生品,生胡芦巴和酒胡芦巴水层萃取物对α-葡萄糖苷酶作用为非竞争可逆。生胡芦巴和酒胡芦巴的乙酸乙酯层和正丁醇层对α-淀粉酶有抑制作用,酒制品的抑制作用优于生品,且均为非竞争可逆。结论:胡芦巴经酒制后可在一定程度上增加对α-葡萄糖苷酶及α-淀粉酶活性的抑制作用,酒胡芦巴水层萃取物有开发成α-葡萄糖苷酶抑制剂的价值,酒胡芦巴的乙酸乙酯和正丁醇提取物具有开发成α-淀粉酶抑制剂的价值。     

Anticholinesterase activity of plastoquinones from Sargassum sagamianum: lead compounds for Alzheimer's disease therapy

During the search for anticholinesterase compounds from marine organisms, two known plastoquinones, sargaquinoic acid (1) and sargachromenol (2), were isolated from Sargassum sagamianum. Both compounds showed moderate acetylcholinesterase (AChE) inhibitory activity in a micromole range (IC(50) 23.2 and 32.7 microm, respectively). However, for butyrylcholinesterase (BuChE), a new target for the treatment of Alzheimer's disease (AD), compound 1 showed particularly potent inhibitory activity (IC(50) 26 nm), which is 1000-fold greater than for AChE. Hence, sargaquinoic acid represents an effective and selective inhibitor of BuChE with a potency similar to or greater than the anticholinesterases in current clinical use, making it an interesting potential drug candidate for AD.     

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??OBJECTIVE To research the effect of new L-phenylalanine derivatives on acetylcholinesterase(AChE) activity. METHODS New L-phenylalanine derivatives were synthesized from substituted 2-bromo-1-acetophenones by four steps reaction, and their inhibitory activities on AChE were measured by Ellman method in vitro. RESULTS The evaluation results showed that most derivatives possessed AChE inhibitory effect and the activity of compound 8b was the most potent with an IC₅₀ value of 8.73??10^-6 mol??L^-1, which was more potent than that of rivastigmine; moreover, compound 8b had no inhibitory activities to BuChE. CONCLUSION The inhibitory activities of new L-phenylalanine derivatives on acetylcholinesterase are worth studying further.
     

芥子碱对大鼠脑匀浆和血清中乙酰胆碱酯酶的抑制作用

目的:考察中药单体芥子碱(3,5-二甲基,4-羟基苯丙烯酰胆碱,sinapine)对大鼠血清和大鼠大脑匀浆乙酰胆碱酯酶(acetylcholinesterase,AChE)的体外抑制作用。方法:大鼠眼球取血后断头,取大脑,匀浆,用双缩脲法测定脑组织匀浆总蛋白含量。采用乙酰胆碱脂酶试剂盒和改良Ellman比色法测定AChE的活力,检测芥子碱对大鼠血清和脑匀浆乙酰胆碱酯酶的抑制作用。结果:芥子碱明显抑制大鼠大脑匀浆AChE的活力,而对血浆AChE活性的抑制相对较弱,对脑匀浆和血清AChE的抑制IC₅₀分别为3.66和22.1μmol.L^-1,IC₅₀血清/IC₅₀脑=6.042(倍)。结论:芥子碱能显著抑制脑组织AChE活性,有可能作为中枢乙酰胆碱酯酶抑制剂(acholinesterase inhibi-tors,AChEIs)具有防治阿尔茨海默病(Alzheimer's disease,AD)的应用前景。     

In vitro acetylcholinesterase inhibitory properties of thymol, carvacrol and their derivatives thymoquinone and thymohydroquinone

The aim of this study was to examine in vitro the inhibitory activity exerted by the main constituents of essential oil obtained from the aromatic plant Thymus vulgaris L. on acetylcholinesterase (AChE). The total essential oil and selected compounds, specifically linalool and thymol, carvacrol and their derivatives thymoquinone and thymohydroquinone, were tested for AChE inhibition. Thymohydroquinone exhibited the strongest AChE inhibitory effect over the range of concentrations. The AChE inhibitory potential decreased in the following order: thymohydroquinone > carvacrol > thymoquinone > essential oil > thymol > linalool. It is interesting that the AChE inhibitory effect exerted by carvacrol was 10 times stronger than that exerted by its isomer thymol, although thymol and carvacrol have a very similar structure.     

Gusanlungionosides A-D, potential tyrosinase inhibitors from Arcangelisia gusanlung

Four new megastigmane glycosides, named gusanlungionosides A-D (1-4), together with 10 known compounds (5-14), were isolated from the stems of Arcangelisia gusanlung. The structures and absolute configurations of 1-4 were elucidated by comprehensive analysis of their NMR and CD data. Compounds 1-4 exhibited strong inhibitory effects not only on the mushroom tyrosinase activity in vitro but also on melanogenesis in cells.     

Labdane-type diterpenoids from Leonurus heterophyllus and their cholinesterase inhibitory activity

In the course of screening plants used in natural medicines as memory enhancers, a 70% ethanol extract of the aerial parts of Leonurus heterophyllus showed significant AChE inhibitory activity. Bioassay-guided fractionation and repeated column chromatography led to the isolation of a new labdane-type diterpenoids (1), named leoheteronin F, and six known compounds (2-7). The chemical structures of isolated compounds were elucidated based on extensive 1D and 2D NMR spectroscopic data. The isolates 1-7 were investigated in vitro for their anticholinesterase activity using mouse cortex AChE enzyme. Leoheteronin A (5) and leopersin G (7), which possess a 15,16-epoxy group at the side chain, were found to be potent in the inhibition of AChE.     

Tabernaemontana divaricata extract inhibits neuronal acetylcholinesterase activity in rats

The current pharmacotherapy for Alzheimer's disease (AD) is the use of acetylcholinesterase inhibitors (AChE-Is). A previous in vitro study showed that Tabernaemontana divaricata extract (TDE) can inhibit AChE activity. However, neither the AChE inhibitory effects nor the effect on neuronal activity of TDE has been investigated in vivo. To determine those effects of TDE in animal models, the Ellman's colorimetric method was implemented to investigate the cortical and circulating cholinesterase (ChE) activity, and Fos expression was used to determine the neuronal activity in the cerebral cortex, following acute administration of TDE with various doses (250, 500 and 1000 mg/kg) and at different time points. All doses of TDE 2 h after a single administration significantly inhibited cortical AChE activity and enhanced neuronal activity in the cerebral cortex. The enhancement of Fos expression and AChE inhibitory effects in the cerebral cortex among the three TDE-treated groups was not significantly different. A 2 h interval following all doses of TDE administration had no effect on circulating ChE activity. However, TDE significantly inhibited circulating AChE 10, 30 and 60 min after administration. Our findings suggest that TDE is a reversible AChE-I and could be beneficial as a novel therapeutic agent for AD.     

Phenolic and triterpene glycosides from the stems of Ilex litseaefolia

Chemical investigation on the stems of Ilex litseaefolia afforded four new phenolic glycosides, litseaefolosides A-D (1-4), and two new triterpene glycosides, spathodic acid 28-O-beta-d-glucopyranoside (5) and (20S)-niga-ichigoside F1 (6), along with 28 known compounds. The structures of 1-6 were determined on the basis of chemical and spectroscopic evidence. Litseaefoloside C (3) showed inhibitory activities in vitro for alpha-glucosidase and lipase with IC(50) values of 34.0 and 0.31 microg/mL, respectively.     

具有乙酰胆碱酯酶抑制活性的总生物碱提取物的筛选

目的:从采自甘肃文县和舟曲的22种植物的总生物碱提取物中筛选具有乙酰胆碱酯酶抑制活性的提取物,为进一步寻找天然乙酰胆碱酯酶抑制剂单体成分奠定基础。方法:采用酸提碱沉法提取生物碱,用改良的Ellman比色法测定总生物碱的抗乙酰胆碱酯酶活性。结果:弯柱唐松草(Thalictrumuncinulatum Franch.)、阔叶十大功劳(Mahnia bealei(Fort.)Steam.)、蚬壳花椒(Zanthoxylum dissitumHemsl.)、刺柄南星(Arisaema aspeiatum N.E.Brown)、一把伞南星(Arisaema erunescens(Wall.)Schott)、猫儿屎(Decaisnea insignis(Griff)Hook.f.et thoms)、虎耳草(Saxifraga stolonifera curt.)和西固凤仙花(Impot-iens notolopha Maxim.)的总生物碱提取物具有明显的乙酰胆碱酯酶抑制活性,其IC50值分别为2.3μg.mL-1、2.9μg.mL-1、3.5μg.mL-1、11.0μg.mL-1、16.9μg.mL-1、22.7μg....