Comparative pharmacokinetics of six major compounds in normal and insomnia rats after oral administration of Ziziphi Spinosae Semen aqueous extract

Ziziphi Spinosae Semen(ZSS),a traditional Chinese medicine,is used in clinics for the treatment of insomnia in China and other Asian countries.Herein,we described for the first time a comparative pharmacokinetics study of the six major compounds of ZSS in normal control(NC)and para-chlorophenylalanine(PCPA)-induced insomnia model(IM)rats that were orally administered the aqueous extract of ZSS.An ultra-high-performance liquid chromatography coupled with quadrupole orbitrap mass(UHPLC-Q-Orbitrap-MS)method was developed and validated for the simultaneous determination of coclaurine,magnoflorine,spinosin,6000-feruloylspinosin,jujuboside A(JuA),and jujuboside B(JuB)in ZSS in rat plasma.The established approach was successfully applied to a comparative pharmacokinetic study.The systemic exposures of spinosin and 6000-feruloylspinosin were decreased in the IM group compared to the NC group,while plasma clearance(CL)was significantly increased.The Tmax values of JuA and JuB in IM rats were significantly lower than those in NC rats.The T1/2 of JuA in the IM group was significantly accelerated.The pharmacokinetic parameters of coclaurine and magnoflorine were not evidently affected between the two groups.These results indicate that the pathological state of insomnia altered the plasma pharmacokinetics of spinosin,6000-feruloylspinosin,JuA,and JuB in the ZSS aqueous extract,providing an experimental basis for the role of ZSS in insomnia treatment.The comparative pharmacokinetics-based UHPLC-Q-Orbitrap-MS using full-scan mode can therefore provide a reliable and suitable means for the screening of potentially effective substances applied as quality markers of ZSS.

Quality analysis of commercial samples of Ziziphi spinosae semen(suanzaoren) by means of chromatographic fingerprinting assisted by principal component analysis

Due to the scarcity of resources of Ziziphi spinosae semen （ZSS）, many inferior goods and even adulterants are generally found in medicine markets. To strengthen the quality control, HPLC fingerprint common pattern established in this paper showed three main bioactive compounds in one chromatogram simultaneously. Principal component analysis based on DAD signals could discriminate adulterants and inferiorities. Principal component analysis indicated that all samples could be mainly regrouped into two main clusters according to the first principal component （PC1, redefined as Vicenin II） and the second principal component （PC2, redefined as zizyphusine）. PC1 and PC2 could explain 91.42%of the variance. Content of zizyphusine fluctuated more greatly than that of spinosin, and this result was also confirmed by the HPTLC result. Samples with low content of jujubosides and two common adulterants could not be used equivalently with authenticated ones in clinic, while one reference standard extract could substitute the crude drug in pharmaceutical production. Giving special consideration to the well-known bioactive saponins but with low response by end absorption, a fast and cheap HPTLC method for quality control of ZSS was developed and the result obtained was commensurate well with that of HPLC analysis. Samples having similar fingerprints to HPTLC common pattern targeting at saponins could be regarded as authenticated ones. This work provided a faster and cheaper way for quality control of ZSS and laid foundation for establishing a more effective quality control method for ZSS.

Quality control of Semen Ziziphi Spinosae standard decoction based on determination of multi-components using TOF-MS/MS and UPLC-PDA technology

A sensitive,fast and comprehensive method for the quality assessment of Semen Ziziphi Spinosae(SZS)standard decoction with characterization of its chemical components was developed and validated.UPLC-Q/TOF-MS/MS system was used to identify thirty-six chemical components of SZS standard decoction which included nucleosides,phenolic acids,alkaloids,and flavonoids.Furthermore,a UPLC-PDA method was validated to simultaneously determine adenosine,protocatechuic acid,magnoflorine,catechin,protocatechin,vicenin II,spinosin,kaempferol-3-rutinoside,and 6'''-feruloylspinosin which represent four species of characteristic compounds.The qualitative method had been validated according to Chinese Pharmacopoeia(2015 edition)in terms of lineary,repeatability,recovery and stability for all analytes,with the results showing good precision,accuracy and stability.In conclusion,the method using UPLC combined with MS and PDA provided a novel way for the standardization and identification of SZS standard decoction,and also offered a basis for qualitative analysis and quality assessment of the preparations for SZS standard decoction.

Primary Study of Novel Method for Determination of Saponin in Ziziphi Spinosae Semen (<i>Suan-Zao-Ren</i>) by Digitally Scanning the Photo of the HPTLC Image without Configuration of TLC Scanner Facility

Conventional quantity determination of Chinese herbal medicines by High-Performance Thin-Layer Chromatography (HPTLC) must be carried out on a TLC scanner for measuring the marker component on the HPTLC plate. Generally the HPTLC image of the herbal drug always takes the photograph as the permanent visual document. Considering the photo of the HPTLC image preserving all the visual quality-related parameters, there might be an alternative cost/effective approach worth studying. It would be presumably applicable to use the exclusively developed software as a virtual scanner instead of the TLC Scanner instrument to do digital scanning, integrating and calculating target components in the photo of the HPTLC image. It means digitally scanning the photo of the HPTLC image solely with software does the same thing as the instrumental scanner does. Based on this protocol, we primarily conducted the assay of the contents of the major saponins, jujuboside a and jujuboside b, in Ziziphi Spinosae Semen (Suan-Zao-Ren). Using home-developed software “digiscan” for acquiring the raw data and integrating the peaks area of the target components with “Origin Pro”, the contents of jujuboside a and jujuboside b can then be calculated respectively. The primary investigated experiment results were well compatible with official HPLC-ELSD method in Chinese Pharmacopoeia. The methodology validation and comparative analysis with HPLC confirmed that the unprecedented method is acceptable. The advantage of the recommended approach is far more simple, flexible and cost/effective than the conventional method, and it can be done at anytime and anywhere once only if the high quality HPTLC image photo is available. There would be a bright perspective for application to other herbal medicines. And the one-stop software for merging digital scanning, integration and calculation together is in progress.

基于UPLC-Q-TRAP-MS分析养心安神药对酸枣仁-五味子配伍煎煮前后10个指标成分含量变化

目的分析养心安神药对酸枣仁-五味子配伍煎煮前后主要指标成分差异变化,并建立含量测定方法。方法采用Welch Ultimate XB-C_(18)色谱柱(2.1 mm×50 mm,5μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相,流速0.4 mL/min柱温40℃,梯度洗脱;质谱采用电喷雾离子源(ESI),多反应监测离子扫描模式(MRM)。分别对该药对单煎液、单煎合并液与合煎液中主要指标成分酸枣仁皂苷A、斯皮诺素、五味子醇甲、五味子酯甲、五味子甲素、五味子乙素、芦丁、槲皮素、山柰酚、山柰酚-3-O-云香糖苷进行含量测定,并分析药对配伍前后主要指标成分的含量变化差异。结果以上10种成分在一定浓度范围内均呈现良好的线性关系;精密度、重复性和稳定性RSD均良好;平均加样回收率在95.0%~102.1%,RSD为0.03%~3.7%。本次实验结果与前期试验认为酸枣仁与五味子在合煎后,主要指标成分降低保持一致性。结论多指标成分同时测定为该药对配伍质量全面控制提供新的方法并为其药效物质基础的深入研究提供依据。

大孔树脂吸附法纯化酸枣仁总黄酮的工艺研究

利用7种大孔树脂对酸枣仁的总黄酮进行纯化,依据其吸附能力及解吸能力,选出最佳的大孔树脂型号,研究了上样液浓度、上样速度、上样液体积对大孔树脂吸附率的影响以及洗脱剂类型、洗脱剂浓度、洗脱速度、洗脱剂体积对大孔树脂解吸率的影响,采用正交试验对酸枣仁总黄酮的纯化工艺进行了优化。试验结果表明,DM301大孔树脂纯化酸枣仁总黄酮效果最佳,在上样液浓度为0.1 mg/mL、上样速度为1 BV/h、上样液体积为30 mL、洗脱剂为丙酮、洗脱剂浓度为100%、洗脱速度为2 BV/h、洗脱剂体积为55 mL的条件下,获得的酸枣仁总黄酮纯度最高,相较于纯化前提高了约1.1倍。

2000—2022年酸枣仁研究进展与趋势可视化分析

目的 探讨酸枣仁近20年的研究热点与发展趋势,为相关研究提供参考。方法 检索CNKI、Web of Science核心合集数据库收录的2000年1月1日—2022年12月31日酸枣仁研究文献。利用VOS viewer对英文文献被引频次及研究机构、关键词热点进行可视化分析;利用CiteSpace软件对中文文献研究机构、作者、突现关键词、关键词叠加时间进行可视化分析,利用Microsoft Excel 2021软件对中、英文文献的年度发表趋势、发文量及中文文献下载频数进行趋势分析。结果 共纳入中文文献4 872篇、英文文献128篇,年发文量整体呈上升趋势。中、英文文献发文最多的研究机构分别是山东中医药大学、天津商业大学,发文最多的作者分别是杜晨晖、XIE Junbo。中文文献高频关键词有“酸枣仁”“镇静”“复方治疗应用”,英文文献高频关键词有“performance”“oxidative stress”。结论 2000—2022年酸枣仁研究热点主要集中在酸枣仁化学成分、药理作用与临床应用分析、配伍研究、组方与制剂方面,酸枣仁的质量控制与评价和酸枣仁防治失眠机制的研究可能成为未来的研究方向。

酸枣仁种皮和种仁化学成分的定性及半定量分析

酸枣仁为鼠李科植物酸枣Ziziphus jujuba Mill.var.spinosa(Bunge)Hu ex H.F.Chou的干燥成熟种子,其分为种皮、种仁两个部位,比较研究酸枣仁不同部位化学成分组成及相对含量可为合理开发利用中药酸枣仁资源提供理论依据。基于超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF/MS)技术,从种皮、种仁中共鉴定出57个化学成分。结合主成分分析(PCA)和正交偏最小二乘法判别分析(OPLS-DA)对两者进行差异成分研究,以变量投影重要度(VIP)值>5为标准,筛选了差异成分17个,其中白桦脂酸、桦木酮酸、麦珠子酸和酸枣仁皂苷Ⅰ主要存在于种皮部位,斯皮诺素、酸枣仁皂苷A和6-阿魏酰斯皮诺素等13个化合物主要存在于种仁部位。通过超高效液相色谱-电雾式检测器(UPLC-CAD)结合反梯度补偿技术,建立半定量液相色谱指纹图谱,考察了6个不同结构类型的代表成分的响应一致性,其不同浓度下平均响应因子间的RSD值为7.04%,各化合物响应一致性良好,可用于酸枣仁的半定量表征分析;结果表明:种皮部位主要成分为白桦脂酸和油酸,其中白桦脂酸的含量约是种仁的7倍;种仁部位主要成分为斯皮诺素、酸枣仁皂苷A、亚油酸、白桦脂酸和油酸,其中斯皮诺素、酸枣仁皂苷A的含量分别是种皮的18倍和24倍。综上,本研究阐明了酸枣仁种皮、种仁的化学成分差异,明确了酸枣仁两个部位中各自的主要成分及其相对含量,为酸枣仁不同部位合理开发和利用奠定了基础。

多指标结合AHP-CRITIC优化酸枣叶拌蒸酸枣仁炮制工艺

目的:采用正交设计结合层次分析法-客观赋权法(AHP-CRITIC)优化酸枣叶拌蒸酸枣仁的炮制工艺。方法:采用单因素试验优化辅料酸枣叶的加入倍量和蒸制时间,以酸枣叶的加入倍量和蒸制时间为因素,以斯皮诺素、6‴-阿魏酰斯皮诺素、酸枣仁皂苷A、白桦脂酸含量为评价指标进行综合加权评分,结合AHP-CRITIC优化酸枣仁的拌蒸工艺。结果:最佳拌蒸工艺为酸枣叶加入量是酸枣仁的4倍,1层酸枣叶1层酸枣仁交替平铺,上锅蒸制3 h。结论:多指标结合AHP-CRITIC优选酸枣仁拌蒸工艺合理可行,可为酸枣叶拌蒸酸枣仁炮制工艺的生产实践提供参考。

酸枣仁炮制前后镇静催眠作用比较研究

目的基于网络药理学和动物实验探讨中药酸枣仁(Semen Ziziphi Spinosae,SZS)炮制前后镇静催眠作用差异。方法通过相关平台和数据库收集SZS的化学成分,检索失眠和睡眠障碍的靶点进行检索,构建蛋白质相互作用关系,绘制出中药-成分-靶点-通路网络,并进行富集分析。结合预测结果,通过戊巴比妥诱导睡眠试验、行为学分析、脑组织相关基因mRNA水平和血浆神经递质评价SZS不同制品对昆明小鼠的影响。结果SZS与失眠相关的靶点基因共有22个,包括乙酰胆碱酯酶(acetylcholinesterase,ACHE)、α-1B肾上腺素能受体(alpha-1B adrenergic receptor,ADRA1B)、钠依赖性5-羟色胺转运蛋白[solute carrier family 6(neurotransmitter transporter,serotonin),member 4,SLC6A4]等,它们可能通过9个相关途径和69个生物学过程发挥作用,其中包括钙信号通路、唾液分泌等。体内实验发现,SZS生用、炒用及生炒合用均能减少小鼠活动性,增强戊巴比妥钠催眠效果,降低皮质及海马中ACHE、ADRA1B和SLC6A4 mRNA水平,调节血浆中乙酰胆碱、肾上腺素等含量,且效果与药物剂量呈正比。结论SZS具有镇静催眠作用,以高剂量的生炒SZS合煎液效果更佳,这种作用可能是由影响神经递质及其相关受体水平,介导中枢神经系统导致的。

酸枣仁与理枣仁鉴别方法研究

目的 建立酸枣仁及常见伪品理枣仁的鉴别方法,为酸枣仁的质量控制和市场监督提供依据。方法 对酸枣仁和理枣仁进行性状及显微鉴别,采用超高效液相色谱串联质谱(UPLC-MS/MS)法测定其中6’’’-芥子酰斯皮诺素的含量,色谱柱为Phenomenex C18柱(100 mm×2.1 mm,2.6μm),流动相为0.1%甲酸-10 mmol/L甲酸铵水溶液-甲醇(梯度洗脱),流速为0.2 mL/min,柱温为30℃,进样量为5μL,离子源为电喷雾离子源,正离子、多反应监测模式扫描,电喷雾电压为3.5 kV,离子源温度为150℃,去溶剂气温度为500℃,扫描范围为50~1 000 amu。结果 酸枣仁和理枣仁在性状上外种皮纹理存在差异,在显微特征上外种皮栅状细胞、种皮内表皮细胞存在差异。6’’’-芥子酰斯皮诺素的质量浓度在0.205 2~6.156μg/mL范围内与峰面积积分值线性关系良好(r=0.997 5,n=6),检测限为21.13 pg。酸枣仁和理枣仁样品的精密度、稳定性、重复性试验结果的RSD均低于3.0%(n=6);加样回收率分别为97.01%和97.80%,RSD分别为2.15%和2.09%(n=6)。10批酸枣仁和理枣仁中6’’’-芥子酰斯皮诺素的含量分别为0.23,3.70μg/mL。拟订酸枣仁中掺伪理枣仁限度为5%,样品中6’’’-芥子酰斯皮诺素测得含量大于0.5μg/mL时可视为掺伪理枣仁。结论 所建立的方法操作简便、结果准确、重复性和稳定性均良好,可有效鉴别酸枣仁和理枣仁。

基于多指标综合分析酸枣仁最佳采收期的质量标志物研究

目的:从酸枣仁出仁率、出油率以及13个主要化学成分含量综合分析其最佳采收期,并确定最佳采收期的质量标志物。方法:采摘8月28日~10月17日酸枣,计算出仁率和出油率。采用GC-MS法以及HPLC-UV-ELSD法测定不同采收期酸枣仁中主要化学成分的含量变化。采用多元统计分析方法筛选出不同采收期酸枣仁差异性成分,通过网络药理学寻找其对应靶点和通路。结果:9月30日采收的酸枣仁出仁率和出油率达到最高值;9月16日到10月17日初级代谢产物(脂肪酸类)和次级代谢产物(生物碱类、皂苷类和黄酮类)含量最高,最终确定9月16日到10月17日为酸枣仁的最佳采收时间。采用多元统计分析、网络药理学分析及结合质量标志物原则,预测5个成分亚油酸、油酸、乌药碱、酸枣仁皂苷A和酸枣仁皂苷B为研究酸枣仁最佳采收期的质量标志物。结论:本研究建立了不同采收期酸枣仁质量多指标综合评价方法,确定了酸枣仁最佳采收期的质量标志物,为酸枣仁的质量控制提供参考。

HPLC法测定新乐康片相关药材中8种活性成分含量

建立HPLC法同时测定新乐康片中三味原料药材钩藤、酸枣仁和萝芙木中钩藤碱、异钩藤碱、斯皮诺素、酸枣仁皂苷A、酸枣仁皂苷B、育亨宾、阿义吗啉及利血平等8种活性成分的含量。采用Titank C18(250 mm×4.6mm,5μm)色谱柱,以乙腈-0.4%磷酸水溶液(含0.05%三乙胺)为流动相,梯度洗脱,检测波长204 nm,流速为1.0 mL/min,柱温为30℃。上述三味药材相关成分之间分离度良好,在λ=204 nm条件下,其在分别的线性范围内与响应值之间的线性关系良好(R^(2)≥0.9990);8种活性成分的平均加样回收率98.37%~101.21%,RSD<2%,仪器精密度、供试品溶液的稳定性与方法重复性均良好。建立的HPLC法可一法对钩藤、酸枣仁和萝芙木三味中药材中8种活性成分含量进行测定,为药业在生产新乐康片前期选材上节约了测定时间,提高了生产效率,同时为新乐康片中活性成分HPLC含量测定方法的建立及其质量标准升级提供参考。

酸枣仁治疗卒中后失眠药理机制及研究进展

卒中是常见的脑血管疾病之一,卒中后失眠是卒中后发病率较高的并发症,主要临床表现为卒中后出现的入睡困难、易醒、早醒等症状,其发病机制复杂且尚不完全明确,现有研究显示其与解剖部位、神经生物学因素及机体整体功能状态和社会心理学等因素有关。神经生物学因素中包含炎症介质、神经递质、脑血流量、血容量、下丘脑-垂体-肾上腺(HPA)轴功能等的变化,长期的卒中后失眠严重时会引起脑血管疾病的复发,对患者的基本日常生活产生了很大的影响,中医认为卒中后失眠是中风与不寐的合病,归属于“不寐”范畴。营卫失衡、阴阳失交、气血失调、神失所司均可导致卒中后失眠。而酸枣仁作为临床中常用的中药,具有养心补肝、宁心安神、敛汗、生津止渴之功用。通过对唐宋时期医家及李济仁国医大师、闫咏梅教授等医家的临床用药规律总结,发现酸枣仁是临床上治疗卒中、卒中后睡眠障碍用药频率最高的单味中药之一,通过大量的临床及实验研究发现酸枣仁的养心安神功效确切,现代研究发现,酸枣仁的主要成分有三萜皂苷类、黄酮类、生物碱类及脂肪酸类等,其具有镇静催眠、抗抑郁、抗氧化、抗肿瘤、抗心律失常、抗心肌缺血等作用。临床上经常用于治疗失眠、焦虑、抑郁等情感障碍类精神疾病以及心血管疾病,该文对酸枣仁有效成分的药理作用及其治疗卒中后失眠的药理机制进行综述,并对酸枣仁的炮制及药对进行归纳梳理,明确酸枣仁对卒中后失眠的重要作用,以提供卒中后失眠的最佳用药选择。

白术、酸枣仁提取物对金黄色葡萄球菌的抑菌效果研究

试验旨在研究菊科植物白术干燥根茎切片提取物(白术提取物)、鼠李科植物酸枣的种子提取物(酸枣仁提取物)单独和联合作用对金黄色葡萄球菌的抑菌效果。通过微量肉汤法测定抑菌浓度和抑制率;用酶标仪测定相应OD值以检测核酸和蛋白质含量、乳酸脱氢酶(LDH)活力及碱性磷酸酶(AKP)活力,以确定对细菌细胞膜和细胞壁的影响,用十二烷基硫酸钠-聚丙烯酰胺(SDSPAGE)凝胶电泳检测金黄色葡萄球菌菌体总蛋白含量变化;用结晶紫染色法检测对细菌生物膜的抑制作用。结果表明:0.4 mg/mL白术、酸枣仁提取物单独及0.1 mg/mL二者联合作用对金黄色葡萄球菌均具有显著抑菌效果;与对照组相比,白术、酸枣仁提取物单独和联合作用均使菌液上清液OD_(260)、OD_(280)数值和AKP活力显著增加,菌体LDH活力和总蛋白含量降低,抑制了生物膜的形成。说明白术、酸枣仁提取物单独和联合作用均可显著抑制金黄色葡萄球菌,且联合作用抑菌效果显著强于各药物单独作用。

酸枣仁汤加减治疗失眠伴焦虑、抑郁的效果分析

目的:分析酸枣仁汤加减治疗失眠伴焦虑、抑郁的效果。方法:选取2020年4月—2023年4月重庆市忠县人民医院中医科收治的失眠伴焦虑、抑郁患者72例作为研究对象,依据随机数字表法分为对照组和观察组,各36例。对照组采取常规药物治疗,观察组采取酸枣仁汤加减治疗,比较两组治疗效果。结果:治疗后,两组汉密尔顿抑郁量表、汉密尔顿焦虑量表、匹兹堡睡眠质量指数量表评分低于治疗前,且观察组低于对照组,差异有统计学意义(P<0.05)。观察组治疗总有效率高于对照组,差异有统计学意义(P=0.001)。观察组不良反应总发生率低于对照组,差异有统计学意义(P=0.001)。结论:酸枣仁汤加减治疗失眠伴焦虑、抑郁的效果较好,可改善患者焦虑、抑郁情况,提高其睡眠质量,且不良反应发生率低。

王济华运用角药治疗类风湿关节炎经验

王济华教授认为,类风湿关节炎早期多因外感风、寒、湿三邪,三者之间相互促进、互相转化,风寒表邪入里日久或郁而化热;病程日久多虚实夹杂,肝肾亏虚、气血不足,同时兼有寒湿痹阻、痰瘀互结。故王教授治疗痹证早期以祛邪散寒为主,兼顾清热利湿、化痰散结,中晚期则偏于滋养气血、补益肝肾。王济华教授运用角药治疗痹证颇有心得,组方精妙、配伍严谨,常用角药川芎-延胡索-白芍行气活血、柔筋止痛;忍冬藤-络石藤-青风藤清热通络、祛风除湿;半夏-陈皮-竹茹燥湿化痰、理气和中;白术-薏苡仁-茯苓益气健脾、利湿消肿;桑寄生-续断-杜仲滋补肝肾、强筋壮骨;酸枣仁-合欢皮-首乌藤宁心安神、解郁除烦。王教授临证时,追求阴阳调和平衡、气机升降有度,临床用药有药简力专、增效减负之功。

Determination of five neurotransmitters in the rat brain for the study of the hypnotic effects of Ziziphi Spinosae Semen aqueous extract on insomnia rat model by UPLC-MS/MS

Ziziphi Spinosae Semen(ZSS) has been used for treatment of insomnia in China for centuries. To reveal the influence of insomnia on the levels of the neurotransmitters including serotonin(5-HT), glutamic acid(Glu),γ-aminobutyric acid(GABA),noradrenaline(NE) and dopamine(DA), and to study the role of ZSS aqueous extract in the treatment of insomnia, an UPLC-ESIMS/MS method was developed and validated for simultaneous determination of five neurotransmitters in the rat brain. The brain samples were pretreated by one-step direct protein precipitation with acetonitrile. The analytes were detected in positive mode with multiple reaction monitoring(MRM) and the procedure was completed in less than 10 min. The method showed a good linearity(R^2 >0.9967) with the other validation parameters were within acceptance range. The results indicated that the concentration of 5-HT, GABA and DA is significantly lower(P < 0.01) in para-chlorophenylalanine(PCPA)-induced insomnia rat model group, while Glu and NE significantly higher than those in control group(P < 0.01). Treatment with ZSS aqueous extract(4 or 8 g·kg^1·d^-1 for seven days) could ameliorate the symptoms of insomnia by significantly changing the levels of the neurotransmitter parameters mentioned above. The data obtained in this study demonstrate that ZSS aqueous extract could ameliorate the symptoms of insomnia by modulating the levels of monoamines and amino acid neurotransmitters in the brain.

不同比例酸枣仁-茯苓水提物对大鼠焦虑样行为的影响

目的:观察不同比例酸枣仁-茯苓混合后水提物的抗焦虑作用并探讨其机制。方法:64只SD大鼠随机分成8组:正常对照组、模型组、帕罗西汀组、酸枣仁组、茯苓组、酸枣仁-茯苓1:1组、酸枣仁-茯苓2:1组、酸枣仁-茯苓3:1组。28 d慢性束缚加孤养方法建立大鼠焦虑症模型,动态观察动物基本形态及体质量变化,造模第8 d起连续给药21 d。高架十字迷宫及旷场实验评价大鼠焦虑样行为;酶联免疫吸附法及荧光实时定量PCR检测大鼠血清、海马及杏仁核组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)含量及m RNA表达的相对量。结果:与模型组相比,不同比例酸枣仁-茯苓均能有效改善大鼠形态、体质量、焦虑样行为并调节血清及组织炎症因子水平(P<0.05,P<0.01)。其中酸枣仁-茯苓3:1复配使用可极显著提高动物造模28 d后体质量(P<0.01),提高高架十字迷宫总穿臂次数、进入开放臂次数比例、开放臂停留时间比例(P<0.01),提高旷场水平运动次数、垂直运动次数(P<0.01),降低大鼠血清、海马及杏仁核组织中炎症因子TNF-α、IL-1β、IL-6水平(P<0.01)及mRNA表达(P<0.05,P<0.01),效果优于酸枣仁-茯苓1:1或1:2复配及酸枣仁或茯苓单独使用。结论:酸枣仁-茯苓复配使用能有效改善焦虑模型大鼠形态、体质量及行为学表现,且酸枣仁-茯苓3:1配比效果最佳。其抗焦虑作用机制可能与降低外周与中枢炎症水平有关。

HPCE法测定酸枣仁中斯皮诺素的含量

建立了一种以硼酸钠溶液为背景电解质,高效毛细管电泳法(HPCE)测定酸枣仁中斯皮诺素含量的方法。考察了缓冲溶液浓度,pH值,分离电压对分离条件的影响,确定了30 mmol/L、pH=9的硼酸钠溶液为缓冲液,分离电压为15 kV的最优测定条件。检测波长为254 nm,进样时间为20 s,在5~200μg/mL范围内具有良好的线性关系,检出限为0.75 mg/L。对16个品系的酸枣仁中斯皮诺素含量进行了测定,结果表明此方法准确可靠,精准度高,稳定性好,可用来测定酸枣仁中的斯皮诺素含量。