The Effect and Mechanism of Fructus lycii on Improvement of Exercise Fatigue Using a Network Pharmacological Approach with in vitro Experimental Verification

Objective This study aimed to investigate the effect and underlying mechanism of Fructus lycii in improving exercise fatigue.Methods A network pharmacological approach was used to explore potential mechanisms of action of Fructus lycii.Skeletal muscle C2C12 cells and immunofluorescence were employed to verify the effect and mechanism of the representative components in Fructus lycii predicted by network pharmacological analysis.Results Six potential active components,namely quercetin,β-sitosterol,stigmasterol,7-Omethylluteolin-6-C-beta-glucoside_qt,atropine,and glycitein,were identified to have potency in improving exercise fatigue via multiple pathways,such as the PI3K-Akt,neuroactive ligand-receptor interaction,IL-17,TNF,and MAPK signaling pathways.The immunofluorescence results indicated that quercetin,a significant active component in Fructus lycii,increased the mean staining area of 2-NBDG,TMRM,and MitoTracker,and decreased the area of CellRox compared to the control.Furthermore,the protein expression levels of p-38 MAPK,p-MAPK,p-JNK,p-PI3K,and p-AKT markedly increased after quercetin treatment.Conclusion Fructus lycii might alleviate exercise fatigue through multiple components and pathways.Among these,quercetin appears to improve exercise fatigue by enhancing energy metabolism and reducing oxidative stress.The PI3K-AKT and MAPK signaling pathways also appear to play a role in this process.

A Rapid Determination of 30 Prohibited Pesticide Residues in Lycii Fructus by UPLC-MS/MS

Prohibited pesticide residues have become one of the main factors affecting the quality and safety of Lycii Fructus,However,rarely studies focus on the rapid determination of these residues.Here,a total of 30 kinds of prohibited pesticide residues were determined by ultra-performance liquid chromatography tandem triple quadrupole mass spectrometry(UPLC-MS/MS)in five different process ways.Pretreatment methods,chromatographic separation and detection conditions in mass spectrometry were all optimized accordingly.Among the five different pretreatment methods,the first and third solid phase extraction failed to provide high recoveries of sulfosulfuron compounds(both lower than 60%).Recovery of chlorphenamidine by the Quick Easy Cheap Effective Rugged and Safe multiresidue method(QuEChERS)was lower than 60%,which did not meet the requirements of trace determination.The concentrations of 30 prohibited pesticides residues treated by straightforward and solid phase extraction showed good linearity in their corresponding ranges,with correlation coefficients over 0.99.The average recoveries of straightforward ranged from 78.13%to 110.9%,while RSD ranged from 1.3%to 16.9%,albeit poor purification was observed.The recovery yield from solid phase extraction was between 67.75%and 103.08%with RSD value from 0.8%to 14.0%,which met the requirements of trace determination,this method has good precision and stability.These results could be employed to other Traditional Chinese Medicines(TCMs)in detecting prohibited pesticide residues.

The Inhibitory Effect of Extracts From Fructus lycii and Rhizoma polygonati on in vitro DNA Breakage by Alternariol

Alternariol caused DNA single-strand breakage. Conversion of the closed circular double-stranded supercoiled DNA (pBR 322) to the nicked circular form and linear form was used to investigate the effect of extracts of some Chinese medical herbs on DNA nicking induced by alternariol. Some substances in the extracts of Rhizoma polygonati (RP) and Fructus lycii (FL) were shown to protect DNA from the attack by alternariol.Some substance in the RP may bind to plasmid DNA, and this binding reduces the electrophoretic mobility of DNA. These results indicate that substances from FL and RP may be used as DNA protectors. It is possible that they play an important role in preventing cancer.

A Network Pharmacology Approach to Uncover the Molecular Targets and Associated Potential Pathways of Lycii Fructus for the Treatment of Retinitis Pigmentosa

Objective To explore the molecular targets and associated potential pathways of Lycii Fructus(LF,Gou Qi Zi,枸杞子)in the treatment of retinitis pigmentosa(RP)by the approaches of network pharmacology and bioinformatics.Methods The potential blood-entry active ingredients and targets of LF were retrieved by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).RP-related gene targets were retrieved through disease comprehensive databases.Protein-protein interaction(PPI)network of LF component-targets and RP disease-targets was constructed by STRING,and the intersection of the 2 networks was extracted.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis of theintersection network were conducted by Database for Annotation,Visualization and Integrated Discovery(DAVID).CytoHubba was used to screen the key targets.Results A total of 188 chemical constituents related to LF was retrieved from TCMSP database.45 active ingredients were screened according to pharmacokinetic parameters oral bioavailability(OB)and drug similarity(DL).36 active ingredients were further screened and 201 targets related to these constituents were obtained.206 target genes directly related to RP were obtained from the disease comprehensive databases,and 89 genes were obtained from the intersection of componenttarget and disease-target PPI network.These genes were mainly involved in intracellular signal transduction,GTPase activity regulation,cell morphology regulation,and other biological processes.Molecular functions were mainly related to Rho guanine nucleotide exchange factor activity,GTPase activator activity,receptor signal protein serine/threonine kinase activity and so on.They were enriched in the cytoplasm,cell membrane,Golgi apparatus,and other regions.The mechanism was related to cell cycle pathways,neurotrophin signaling pathways,Ras signaling pathways,and so on.10 key gene targets of LF in the treatment of RP were screened.Conclusions The material basis for LF to exert its pharmacodynamic effect is 36 active ingredients such as cycloartenol,mandenol,and so on.The key targets of LF in the treatment of RP include 10 genes,such as Rho,PAK,and so on.The main mechanism is related to the regulation of the Ras signaling pathway,neurotrophin signaling pathway,cell cycle related pathway,and other signaling networks.

Effects of Lycii Fructus and Salviae Miltiorrhizae on the Syndrome of Deficiency with Blood Stasis in RCS(rdy-/-, p-/-) Rats with Retinitis Pigmentosa: An Intervention Study

Objective To investigate the effects of Lycii Fructus(LF,Gou Qi Zi,枸杞子)and Salviae Miltiorrhizae Radix Ex Rhizoma(SM,Dan Shen,丹参)on the syndrome of deficiency with blood stasis in the RCS(rdy-/-,p-/-)rats with retinitis pigmentosa(RP).Methods A total of 32 RCS(rdy-/-,p-/-)rats were divided into 4 groups(equal amounts of female and male rats in each group):model group treated with 0.9%normal saline,LF group treated with LF formula granules,SM group treated with SM formula granules,and LF and SM(L·S)group treated with LF and SM formula granules.Eight RCS(rdy+/+,p+/+)rats(4 males and 4 females)were treated with 0.9%normal saline to serve as blank group.The contents of E2,PG,P-Selectin,plasma viscosity,whole blood relative index of the high shear rate and fibrinogen content in plasma,and the content of cAMP and cGMP in retinal homogenate were detected.The retina was evaluated by hematoxylin-eosin staining.Results The contents of E2,PG,P-Selectin,plasma viscosity,whole blood relative index of the high shear rate,and fibrinogen content in the plasma of L·S group significantly differed from those of model group(P<0.01),but were similar to those of blank group.The contents of cAMP and cGMP in the retinal homogenate of L·S group significantly differed from those in model group(P<0.01)but were similar to those in blank group(P>0.05).Conclusions LF and SM can effectively treat retinitis pigmentosa by ameliorating the syndrome of deficiency with blood stasis.

基于外观性状和指标性成分划分枸杞子不同产地

目的 对不同产地枸杞子外观性状与其指标性成分进行分析,并探究两者是否存在关联性,为进一步研究枸杞子质量评价提供方法。方法 依据《中华人民共和国药典》2020版规定对枸杞子中甜菜碱、枸杞多糖的含量进行测定;对枸杞子的外观性状进行测量统计,采用逐步判别法、线性判别分析法(LDA)对枸杞子外观性状数据进行筛选并优化;并分析枸杞子外观性状与其指标性成分之间的关联性。结果 宁夏枸杞多糖含量高达3.43%,远高于其他产地。对枸杞子外观性状与内在品质进行关联性分析,发现果皮厚度与甜菜碱含量呈负相关性(r=-0.485,P<0.01);纵横比与枸杞多糖含量(r=0.420,P<0.05)、甜菜碱含量(r=0.815,P<0.01)呈正相关性。结论 不同产地的枸杞子在外观性状及内在指标上均有差异,该研究可为枸杞子的综合质量评价提供参考。

基于生物信息学探究枸杞子治疗阿尔茨海默病的分子机制

目的基于生物信息学揭示枸杞子治疗阿尔茨海默病(AD)的潜在靶点,探讨其相关生物学机制。方法通过检索中药系统药理学数据库和分析平台(TCMSP)获得枸杞子的活性成分及其潜在靶点,通过基因-疾病关联数据库(DisGeNET)检索AD相关基因靶标。将枸杞子活性成分的潜在靶点和AD相关基因交集后的靶基因,导入功能性蛋白质关联网络数据库(STRING)构建蛋白质-蛋白质相互作用网络,鉴定出关键靶点,进一步构建靶点-通路-疾病的相互作用网络,同时对基因本体论(GO)通路和京都基因与基因组百科全书(KEGG)通路进行富集分析。利用Cytoscape 3.7.1建立成分-靶点-通路-疾病网络。采用AutoDockVina软件进行分子对接并计算结合亲和力。结果从枸杞子中筛选得到36种相关成分,对应靶点202个。从DisGeNET数据库中收集了123个与AD相关的重要基因,最终确定了28个活性枸杞子成分和AD的共享靶点,其中10个靶点:肿瘤坏死因子、白细胞介素(IL)-6、胱天蛋白酶3、血管内皮生长因子A、前列腺素内过氧化物合酶2、超氧化物歧化酶1、IL-1B、淀粉样前体蛋白、过氧化物酶体增殖物激活受体γ、血红素加氧酶1,通过最大团节点中心性算法进一步定义为关键基因靶点。在P<0.01的条件下,GO途径的富集分析分别证明了10个生物学过程、分子功能和细胞成分。KEGG通路富集分析得到102条P<0.05的通路。分子对接显示,槲皮素、甘氨酸、β-谷甾醇等具有较高的亲和力,可以成为治疗AD的潜在药物。结论枸杞子治疗AD的机制与细胞凋亡、神经炎症、神经退行性病变等途径有关,可为枸杞子治疗AD提供药理学依据。

三补方对自然衰老小鼠认知功能及脂质过氧化损伤的影响

目的观察三补方对自然衰老小鼠认知功能的影响,探讨潜在病理因素和三补方(Sanbu Decoction,SBD)的作用及其可能作用机制。方法18只18月龄自然衰老健康SPF级野生型C57BL/6小鼠,雌雄各半,随机分为衰老组(WT-Aged)、多奈哌齐组(WT-Donepezil)、三补方组(WT-SBD),另选6只3月龄小鼠为青年对照组(WT-Young)。WT-Young小鼠不灌胃,WT-Aged小鼠用蒸馏水灌胃,其余用相应药物处理,灌胃45 d。灌胃结束前6日采用水迷宫实验,记录潜伏期时间、穿越平台次数和平台所在象限游泳路程。用HE染色观察海马神经元形态变化,免疫组织化学法测定小鼠脑海马组织Tau、p-Tau、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)、神经上皮干细胞蛋白(neuroepithelial stem cell protein,Nestin)表达水平,检测试剂盒检测超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性和丙二醛(malondialdehyde,MDA)含量。结果与WT-Young相比,第2日起WT-Aged逃逸潜伏期延长(P<0.05),穿越平台次数和平台所在象限经过的路程减少(P<0.05);与WT-Aged相比,WT-Donepezil第3~5日和WT-SDB第4~5日逃逸潜伏期缩短(P<0.05),穿越平台次数和在平台所在象限经过的路程增多(P<0.05)。WT-Aged小鼠出现海马神经元排列紊乱和大面积的神经元坏死,细胞质嗜酸性增强,细胞核固缩、溶解、消失。与WT-Aged小鼠相比,WT-Donepezil和WT-SDB小鼠神经元损伤减少,排列较为紧密,退化固缩的细胞数量则明显减少,少部分细胞可见核固缩及细胞浆嗜酸性变。与WT-Young相比,WT-Aged Tau、p-Tau和GFAP表达增多(P<0.05),Nestin表达减少(P<0.05),SOD、GSH-Px活性降低(P<0.05),MDA含量升高(P<0.05)。与WT-Aged相比,WT-Donepezil和WT-SDB p-Tau、GFAP表达减少(P<0.05),WT-SBD SOD、GSH-Px活性升高(P<0.05),MDA含量降低(P<0.05)。结论三补方可以降低自然衰老小鼠脑海马组织p-Tau表达,减弱星形胶质细胞活化水平,降低脂质过氧化反应,发挥脑保护抗衰老作用,提高老年小鼠学习记忆能力,改善衰老小鼠的衰老状态,延缓衰老进程。

枸杞子抗衰老药理作用研究进展

枸杞子是具有滋补功效的传统药食同源中药,在现代医疗及养生保健等领域得到广泛应用并仍保有巨大的市场开发价值。但目前鲜见关于枸杞子抗衰老作用机制的总结阐述,为明确枸杞子的抗衰老作用,深入解析其分子机制,该研究系统梳理了历代本草对其轻身、耐老的认识及应用,特别是近5年枸杞子抗衰老的国内外相关研究,总结出枸杞子可以通过抗氧化、抗炎、免疫调节、代谢调节、凋亡调节、自噬调节、内质网应激等途径发挥显著的抗衰老作用。该文将为枸杞子抗衰老应用提供科学依据,为充分合理地发挥其抗衰老价值提供参考。

枸杞子-菊花药对提取物对豚鼠光源性近视模型的影响及机制研究

目的考察枸杞子与菊花药对配伍对发育期豚鼠光源性近视发生及发展的影响。方法采用三基色荧光灯照射发育期豚鼠,建立光源性近视模型,枸杞子-菊花提取物添加饲料饲养豚鼠,利用眼科A型超声测量仪检测眼轴,HE染色观察豚鼠视网膜病理损伤,酶联免疫吸附实验(ELISA)检测豚鼠视网膜多巴胺(DA)及褪黑素(MT)含量,转录组学分析等方法探讨枸杞子-菊花对豚鼠光源性近视的影响。结果造模6周后,与正常组相比,模型组豚鼠眼轴显著增长(P<0.01),近视模型建立成功;枸杞子-菊花提取物可有效减缓模型组豚鼠晶状体加厚、玻璃体腔加深,进而延缓整体眼轴的过度增长;模型组豚鼠视网膜组织变薄,视网膜内核层及外核层的厚度及细胞数目显著降低,神经节细胞核稀疏,神经节细胞层空泡化明显,枸杞子-菊花提取物能保护视网膜细胞,视网膜内核层及外核层的厚度及细胞数目增加,神经节细胞核增多;ELISA检测显示模型组豚鼠视网膜内DA、MT水平显著降低(P<0.05),中、高剂量枸杞子-菊花提取物能够有效提升模型组豚鼠视网膜及血清中DA含量(P<0.01,P<0.001);眼部转录组学分析筛选的差异基因主要富集于过氧化物酶体增殖物激活受体(PPAR)信号通路、含胶原蛋白的细胞外基质等相关途径;ELISA结果显示,高剂量枸杞子-菊花提取物能显著增加PPARγ、COL1A1含量(P<0.01),降低SMA含量(P<0.05)。结论枸杞子-菊花药对具有延缓发育期豚鼠光源性近视发生及发展效用,保护视网膜细胞,改善人工光环境导致的DA分泌紊乱,参与调控眼部PPARγ水平,对光源性近视的防控具有积极意义,为临床合理用药和创制眼功能健康产品提供了科学依据。

GC-MS法同时测定枸杞中16种农药残留的含量及安全性评价

目的建立GC-MS法同时测定枸杞中16种农药残留量的含量,并进行安全性评价。方法分析采用DB-5MS色谱柱(30 m×0.25 mm,0.25μm);程序升温;进样量1.0μL,不分流;体积流量1.0 mL/min;进样口温度250℃;电子轰击电离(EI),电子能量70 eV;离子源温度230℃;多反应监测模式;碰撞气高纯N_(2)。采用残留水平、膳食摄入风险、风险排序和累积暴露评估对农药残留膳食风险相对较高的农药进行分析与讨论。结果16种农药在各自范围内线性关系良好(r≥0.9944),平均加样回收率70%~114%,RSD小于2%。产地枸杞和商超枸杞均未出现农药残留超标情况;产地枸杞中氟氯氰菊酯的平均残留量最高,为0.9992 mg/kg,商超枸杞中氯氰菊酯的平均残留量最高,为0.0884 mg/kg;产地枸杞和商超枸杞的慢性和急性膳食风险最高的农药均是氯氰菊酯;溴氰菊酯、哒螨灵、毒死蜱、敌敌畏、杀扑磷风险较高,但实际检测中只有哒螨灵被检出,并且未超标;产地、商超枸杞慢性危害指数(HI)值分别为0.0129、0.0012,急性HI值分别为0.0655、0.0054。结论部分枸杞样品存在农药残留,但均处于安全水平,喷洒农药时需要注意氯氰菊酯的使用。

枸杞子结合多酚碱水解与酸水解提取工艺的比较和产物分析

为了全面评估枸杞子中的酚类成分,本研究分别利用碱水解和酸水解法制备了其中的结合多酚(non-extractable phenolic compounds, NEPC)。选取Box-Behnken实验设计建立了酸或碱的浓度、水解时间和料液比等与NEPC得率之间的数学模型,并优化了水解工艺参数。碱水解过程中的时间对NEPC得率影响显著,而料液比对酸水解的结果影响较大;最佳工艺条件下,碱水解和酸水解获得的NEPC分别为0.65±0.05 mg/100 g和0.52±0.03 mg/100 g。利用UPLC-Q-TOF-MS,从水解产物中解析了11种化合物,主要是苯丙素衍生物。11种化合物都可在酸水解产物中被检出,而碱水解产物中仅鉴定出4种物质。研究结果表明,碱水解能得到更多的NEPC,酸水解产物则表现出良好的化合物多样性,因此制备NEPC时,应根据不同的研究目的选择水解方法。

枸杞不同部位UV-Vis和HPLC指纹图谱研究

目的采用多维指纹图谱结合化学计量学方法,比较枸杞果实和根皮中黄酮、香豆素类和苯丙酸类等成分的整体差异,并综合多糖和总黄酮的含量比较,为枸杞不同药用部位的价值评估和质量标准的建立提供理论依据。方法建立HPLC指纹图谱进行相似度评价和共有峰确定。化学计量学方法对药材不同部位进行分类研究,建立分类模型,筛选出重要差异性标志物并进行定量分析;紫外-可见光谱(UV-Vis),反映药材化学组成的整体特征;采用2020年版《中华人民共和国药典》苯酚-硫酸法测定枸杞多糖的含量,亚硝酸钠-硝酸铝比色法测定总黄酮含量。结果HPLC指纹图谱共标定9个共有峰,指认了绿原酸、东莨菪内酯、槲皮素、芦丁4个色谱峰。相似度评价表明果实和根皮整体组成差异较大,地骨皮不同产地之间质量缺乏一致性;正交偏最小二乘法-判别分析找出芦丁是对整体差异性贡献比较大的化学成分;UV-Vis呈现明显的指纹特性;枸杞子中枸杞多糖和总黄酮的含量分别为2.74%~3.79%和1.52%~2.24%,均高于地骨皮中1.92%~2.67%和0.46%~0.58%,且含量分布具有产地差异性。结论综合多指标揭示枸杞果实和根皮中化学成分差异,为枸杞药材的质量评价和资源价值的充分利用提供重要的数据参考。

基于指纹图谱的化学模式识别法和多指标测定评价枸杞不同部位的差异性

目的研究枸杞植株不同部位中主要化学成分的整体信息和含量差异,综合评价枸杞不同部位的质量和合理应用。方法以3个主产区的枸杞子、枸杞叶和地骨皮药材为研究对象,建立HPLC指纹图谱,采用中药色谱指纹图谱相似度评价系统进行相似度评价;结合SIMCA 14.1软件进行主成分分析等;采用分光光度比色法测定枸杞多糖和总黄酮含量;并利用HPLC方法对甜菜碱进行含量测定。结果枸杞不同部位化学成分的种类和含量均存在明显差异,叶和果实含有更丰富的化学成分,枸杞多糖、总黄酮和甜菜碱的含量均为枸杞叶>枸杞子>地骨皮。结论枸杞叶含有更丰富的营养成分,其药理功能和营养价值可能优于枸杞子。因此,可考虑多产地开发枸杞叶作为枸杞药材来源之一,以促进枸杞资源的多途径、多层次开发利用。

基于UPLC指纹图谱与抗氧化谱效关系的枸杞子质量标志物研究

目的:建立基于超高效液相色谱法(UPLC)指纹图谱与抗氧化谱效关系的枸杞子质量标志物研究方法,为其质量评价提供依据。方法:采用UPLC建立31批枸杞子指纹图谱,基于指纹图谱共有峰峰面积结果,采用聚类分析、主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)方法评价枸杞子整体质量并筛选出主要质量标志物。以多功能氧化酶(MFO)、1,1-二苯基-2-三硝基苯肼(DPPH)、总抗氧化能力(T-AOC)的测定值评价枸杞子的抗氧化能力,利用双变量相关性分析法与正交偏最小二乘法-回归分析研究指纹图谱共有峰与抗氧化活性指标的谱效关系,综合分析筛选质量标志物。结果:枸杞子UPLC指纹图谱共确定34个共有峰,31批枸杞子聚为3类,可区分不同产地的枸杞子;PCA得到4个影响枸杞子分类的主要因子;OPLS-DA显示共有峰2、7、17、19、33的化合物可作为鉴别和区分枸杞子的主要质量标志物;双变量相关性分析表明峰8、11、15、20、23、34的化合物与MFO、DPPH、T-AOC的测定值呈极显著相关,正交偏最小二乘法-回归分析表明峰11、15、23、25、31、33、34的化合物是主要的活性成分群,此类成分可作为枸杞子抗氧化活性的质量标志物。结论:研究结果阐明了枸杞子抗氧化活性的物质基础,筛选出的质量标志物可为枸杞子的质量控制与评价提供依据。

Gouqizi(Fructus Lycii)seed oil reduces D-galactose induced inflammation in testis of rats via Janus kinase 1/signal transducerand activator of transcription 1/nuclear factor-κB in vitro and in vivo

OBJECTIVE:To investigate the efficacy of Gouqizi(Fructus Lycii)seed oil(FLSO)on D-gal induced inflammation in testis of rats in vitro and in vivo.METHODS:In aging Sertoli cells(TM4 cells)induced by D-galactose(D-gal),the expression of upregulated agingrelated proteins.The number of cells counted by cell counting kit(CCK)-8 assay showed a high number of cells disposed with FLSO at 50,100 and 150μg/mL compared to that for the aging model.In vivo,male Sprague–Dawley rats(n=50,8-week-old,230-255 g)were randomly categorized into control,aging model,and FLSO(low-,medium-,and high-dose)groups.The expression of nuclear factor-κB(NF-κB)and its upstream factors[Janus kinase 1(JAK1)and signal transducerand activator of transcription 1(STAT1)]was detected by Western blot and immunofluorescence,related inflammatory factors quantified by enzyme-linked immunosorbent assay.Evaluation of testicular tissue by Johnsen score,the spermatogenic function was explored.RESULTS:The expression of interleukin-1β(IL-1β)(P<0.05),IL-6(P<0.001),and tumor necrosis factorα(TNF-α)(P<0.05)was decreased significantly,while that of heme oxygenase-1(HO-1)(P<0.001)and IL-10(P<0.05)was increased in cells disposed with FLSO 100μg/mL.FLSO inhibited the expression of NF-κB and declined pp65/p65(P<0.01),as detected by Western blotting.In vivo,the levels in serum of IL-1β(P<0.001),IL-6(P<0.05),and TNF-α(P<0.01)declined while IL-10(P<0.05)was upregulated post-FLSO treatment.In addition,the expression of JAK-1 and STAT1 increased significantly in testicular tissue of rats treated with FLSO as compared to the aging model of rats(P<0.001),while the expression of NF-κB(P<0.001)declined in the testis in the FLSO group,as assessed by immunofluorescence.The levels of inhibor B and testosterone in serum both increased(P<0.05).CONCLUSIONS:In conclusion,this study determined the protective effects of FLSO to tolerate inflammatory injury in the testis,indicating that FLSO alleviates inflammation via JAK-1/STAT1/NF-κB pathway.

建立HPLC-RID测定枸杞子中枸杞多糖含量研究

目的通过高效液相色谱-示差折光检测法(HPLC-RlD)摸索枸杞子中枸杞多糖(枸杞多糖以葡萄糖计)的提取方式和含量测定方法,达到检测方法准确可靠操作简便的目的。方法本法采取加水回流提取方式,利用高效液相色谱仪和示差折光检测器进行检测,色谱柱为LC026(Prevail Carbohyrate ES为填充剂250 mm×4.6 mm,5μm),流动相为乙腈-水(70∶30),检测器温度40℃,流速1.0 ml/min,柱温40℃。结果枸杞子在0.5~4.0 mg/ml线性范围内呈良好线性关系,R^(2)=0.9999,平均回收率为104.81%,RSD=1.10%。结论该方法操作简便,灵敏度高,重复性好,准确可靠,可用于测定药材枸杞子中枸杞多糖的含量,并有效控制枸杞子质量。

枸杞子治疗脑卒中潜在作用机制的网络药理学研究

目的利用网络药理学方法探究枸杞子治疗脑卒中的潜在作用机制。方法在TCMSP平台上获取并筛选枸杞子的活性化合物。利用TargetNet数据库预测活性化合物的靶点后,将其与CTD数据库中获得的脑卒中相关靶点取交集得到枸杞子治疗脑卒中的潜在作用靶点。借助String数据库分析潜在作用靶点的蛋白相互作用,并利用Cytoscap e进行拓扑计算,筛选核心靶点。此外,对潜在作用靶点进行了GO功能和KEGG通路富集分析。结果共筛选得到枸杞子活性化合物45个,对应101个靶点。与417个脑卒中相关靶点取交集后得到16个潜在作用靶点。其中核心靶点有7个,分别为:核受体亚家族3C组成员1(NR3C1)、雌激素受体1(ESR1)、雄激素受体(AR)、前列腺素内过氧化物合酶2(PTGS2)、热休克蛋白90α家族A成员1(HSP90AA1)、细胞核因子κB p65蛋白(RELA)、细胞色素P450家族成员17A1(CYP17A1)。主要涉及了氧化还原过程、一氧化氮生物合成过程的正调控、炎症反应的调节等生物过程,富集的通路包括含5-羟色胺的突触、色氨酸代谢、类固醇激素生物合成、苯丙氨酸代谢、组氨酸代谢等。结论枸杞子可能通过调节炎症反应、参与氧化还原过程等发挥神经保护作用,其机制涉及多条信号通路。

GEO芯片分析联合网络药理学探讨五味子-枸杞子保护放射性肝损伤的机制研究

利用转录组学、网络药理学和分子对接的技术预测五味子-枸杞子保护放射性肝损伤的活性成分、作用靶点及作用机制。通过中药系统药理学数据库与分析平台(TCMSP)筛选出五味子-枸杞子主要活性成分,从五味子中筛选出14个活性成分,枸杞子中筛选出25个活性成分,核心成分主要为槲皮素、黄豆黄素和去氧紫草素;通过GeneCards、在线人类孟德尔遗传数据库获得辐射损伤的靶点基因,利用UniProt对所获得的基因进行校正,共同关键靶点共66个,利用Cytoscape 3.8.0软件构建药物-活性成分-关键靶点-放射性肝损伤调控网络,将活性成分与疾病共同靶点上传至String数据库,构建药物靶蛋白-放射性肝损伤蛋白相互作用(PPI)网络,根据度值筛选关键核心靶点为CASP3、EGFR、ESR1;对关键靶点进行功能富集分析及通路富集分析,五味子-枸杞子活性成分主要参与化学致癌-受体激活、脂质与动脉粥样硬、PI3K-Akt、MAPK、凋亡等信号通路;分子对接结果提示3个活性成分与靶蛋白结合能力较强。本研究初步发现五味子-枸杞子可能通过槲皮素、黄豆黄素和去氧紫草素调节化学致癌-受体激活、脂质与动脉粥样硬、PI3K-Akt、MAPK、凋亡等多个信号通路发挥保护放射性肝损伤的作用。

基于生物-化学-形性指标关联的枸杞子综合质量评价研究

目的:基于枸杞子生物-化学-形性指标间的关联分析,筛选能够表征枸杞子功效、浅出枸杞子等级划分标准的核心指标。方法:通过网络药理学方法确定枸杞子功效发挥涉及的生物活性、化学成分,结合本草记载及现有文献、标准确定关联成分的性状指标;对三性指标进行“生物-化学”“化学-形性”“化学-化学”的相关性分析,筛选枸杞子等级划分指标,在此基础上对不同等级的枸杞子核心指标进行差异比较,验证指标的合理性。结果:玉米黄素二酯、β-胡萝卜素均与百粒重、甜度呈显著负相关关系,与枸杞多糖、L*(D65)、纵横比、DPPH清除率呈正相关关系;DPPH清除率与NO抑制率呈正相关关系;百粒重、甜度与果糖呈正相关关系,与枸杞多糖呈负相关关系。结论:β-胡萝卜素、玉米黄素二酯能够关联生物活性、化学成分、形性指标,可作为枸杞子质量的标志性化学成分;百粒重、甜度能够体现不同等级化学成分含量差异,可为枸杞子等级判定浅出标准。