The impact of^(60)Co-γirradiation on the chemical constituents of Chuanxiong Rhizoma

Background:In order to clarify the inmpat ofγirradiation on the chemical composition of traditional Chinese medicine,this paper carefully choosed Chuanxiong Rhizoma to carry on a demonstration study.Methods:Through a meticulous assessment,a comprehensive comparison was made between the irradiated and unirradiated Chuanxiong Rhizoma samples.The property characteristics were investigated by colorimeter and electronic nose.The changes in chemical structures and contents was analyzed by fourier infrared spectroscopy,high performance liquid chromatography and fingerprinting.In a quest to uncover the presence of any new radiolysis products,cutting-edge techniques like ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry and gas chromatography-mass spectrometry were employed.Moreover,the difference of antioxidant activity were investigated.Results:The irradiation doses within 12 kGy had no significant effects on the content of the main chemical components,characteristics and in vitro antioxidant activity of Chuanxiong Rhizoma,while changes in some functional groups and degradation of some volatile oil components containing olefins need further study.Conclusion:This study indicates that^(60)Co-γirradiation is a stable method for sterilization of Chuanxiong Rhizoma.It’s also provide a reference for the establishment of irradiation standards for Chuanxiong Rhizoma and other aromatic medicinal plants.

外源Cd对川芎苓种萌发及幼苗生长的影响

本研究旨在探究不同浓度镉(Cd)对川芎苓种萌发和幼苗生长的影响。采用以石英砂为基质的水培试验,施加不同浓度CdCl 2·2.5 H 2O(0、0.1、0.25、0.5、1、2 mmol/L)溶液处理川芎苓种,研究Cd胁迫对川芎苓种萌发、幼苗生长、生理代谢及Cd含量的影响。结果显示,随着Cd浓度的增加,株高抑制率、根长抑制率上升;根、叶及节盘Cd含量显著升高;株高、根长、叶鲜重、根鲜重下降;抗氧化酶(过氧化氢酶、过氧化物酶、超氧化物歧化酶)活性、非酶系统物质(可溶性蛋白、谷胱甘肽、脯氨酸)含量以及丙二醛含量先升高后降低。研究表明,0.1~2.0 mmol/L Cd对川芎苓种萌发和早期幼苗生长均有明显的毒害作用,随着Cd胁迫浓度的升高毒害作用增强,其中川芎幼苗根对Cd胁迫最敏感。Cd胁迫下川芎幼苗生理生化指标整体上呈现低促高抑的Hormesis效应。川芎幼苗通过提高抗氧化酶活性和非酶系统物质积累,减少膜系统损伤、活性氧蓄积,缓解Cd毒害作用,提高耐受性。

Integrated network pharmacology analysis and in vitro cell experiments to reveal the mechanisms of Ligusticum chuanxiong Hort.in the treatment of non-alcoholic fatty liver disease

Background:Non-alcoholic fatty liver disease(NAFLD)is a liver disease of unknown etiology.A traditional Chinese medicine Ligusticum chuanxiong Hort.(CX),it has been used about 2,000 years.Until now,the mechanism of action of CX on NAFLD remains unclear.Method:We first tested the toxicity of CX to AML12 cells with CCK-8.In vitro cell models of NAFLD were made using free fatty acid,and used Oil Red O staining tested lipid droplets.Then the active compounds of CX were collected from TCMSP and literatures,and SwissTargetPrediction,Search Tool for Interacting Chemicals,Encyclopedia of Traditional Chinese Medicin,Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine database were used to predict the targets of the compounds.DRUGBANK,Online Mendelian Inheritance in Man,Therapeutic Target Database,DisGeNET and GeneCards database were used to predict the targets of NAFLD.Use Venn diagram to obtained the intersection targets by,and analyzed the protein-protein intersection network.Use Kyoto Encyclopedia of Genes and Genomes and Gene Ontology to forecast the function of intersection genes.Molecular docking was used to evaluate the interaction between hub gene and active ingredients.Finally,use western blotting to determine the effects of CX on PPARA,PPARG,IL1B and TNF proteins.Result:CX can reduce the production of AML12 cell lipid droplets.A total of 15 chemical components were identified from CX.Folic acid,chrysophanol and sitosterol were the main components of CX against NAFLD.ALB,TNF,PPARG and PPARA proteins were the main targets of CX in the treatment of NAFLD.PPAR signaling pathway and fatty acid degradation were closely related to anti-NAFLD.Molecular docking results shows that folic acid was the main active ingredient of CX for NAFLD treatment,and TNF is the main potential target.The cellular NAFLD model showed that CX up-regulated the expression of PPARA and PPARG protein and down-regulated inflammatory factor IL-1B and TNF expression.Conclusion:Our study suggests that CX has a therapeutic effect on NAFLDA,which may be related to the PPAR pathway and the reduction of inflammatory cytokines.

Exploring the mechanism of Saposhnikoviae Radix and Chuanxiong Rhizoma treating Parkinson's disease based on network pharmacology and molecular docking

Backgroud:Parkinson’s disease(PD)is a neurodegenerative disorder with an increasing global prevalence.However,the development of drugs for PD treatment has not kept pace with the continuously growing number of patients.Currently,the search for new effective substances from natural drugs is a major research direction.Two Chinese medicinal materials,Saposhnikoviae Radix(Fangfeng)and Chuanxiong Rhizoma(Chuanxiong),are commonly used in the treatment of PD in China.However,the mechanism of their combination is not clear,and further research is needed.Methods:Data were collected from publicly available databases:TCMSP,UnitProt,GeneCards OMIM,PharmGKB,Therapeutic Target Database and DrugBank.Network pharmacology and molecular docking methods was used to analyze the data to discover the possible pharmacological effects of the two drugs in the treatment of PD.Results:Beta-sitosterol,Mandenol and Wallichilide were the active components of Saposhnikoviae Radix and Chuanxiong Rhizoma(FC),and they stably bonded with PD targets,including PTGS2,CASP3,AKT1 and JUN.The target genes of FC were significantly enriched in PD-associated pathways,including calcium signaling and apoptosis pathways.Moreover,the study revealed that the active components of FC may affect cellular structures,such as membrane rafts,membrane microdomains,membrane regions,and postsynaptic membranes,which,in turn,affect a variety of molecular functions and biological processes.Conclusion:The results of this study indicate the direction for clarifying the pharmacodynamic substances of FC,the extraction method of pharmacodynamic substances,as well as the mechanism and efficacy of pharmacodynamic substances.Importantly,this study provides a strategy for developing new therapeutic drugs for PD.

The mechanism of Chuanxiong Rhizoma on glaucomatous optic nerve injury based on network pharmacology and molecular docking

Based on network pharmacology,this study predicted the potential molecular mechanism and related pathways of the protective effect of traditional Chuanxiong Rhizoma,a traditional Chinese herb,on glaucomatous optic nerve injury,and conducted in vitro experimental verification of the predicted results of network analysis.We analyzed the molecular mechanism of Chuanxiong Rhizoma in the potential treatment of glaucoma by revealing its main active ingredients and predicting its targets,so as to provide reference for subsequent basic research.Network pharmacological research results showed that the potential hub targets and key signaling pathways of Chuanxiong Rhizoma in the treatment of glaucoma were closely related to biological processes such as apoptosis,autophagy,inflammation,oxidative stress and angiogenesis.Molecular docking showed that many active ingredients,such as chrysophanol(CHR),myricanone and retinol,could combine well with their target proteins by intermolecular forces,especially CHR had strong binding ability with each target.We speculated that the main active component of Chuanxiong Rhizoma might be involved in the regulation of PI3K-Akt,Nod-like receptor,IL-4 and IL-13,MAPK,AGE-RAGE and neurotrophin signaling pathway by regulating of PI3K,Akt,TLR4,RAGE,NTRK2 and other key targets.Furthermore,it may achieve multi-directional intervention on apoptosis/autophagy,inflammation/immunity,oxidative stress and nutrient metabolism of axoplasma flow,and then delay the degeneration of optic nerve injury.In vitro experiments showed that the active component CHR of Chuanxiong Rhizoma could reverse the M1-type polarization and autophagy/apoptosis of mouse microglia(BV2)induced by lipopolysaccharide(LPS)at the transcriptional level.Meanwhile,the expression of inflammatory mediators IL-1βand TNF-αwas inhibited,and the mRNA level of anti-inflammatory factor IL-10 was significantly increased.In addition,CHR down-regulates activation of the RAGE-NOX4 pathway mediated by LPS in reducing oxidative stress.In this study,network pharmacology and molecular docking technology were integrated for the first time to explore the potential molecular mechanism of traditional Chinese herb“Chuanxiong Rhizoma”in the treatment on glaucoma,and CHR was innovatively proposed as an important ingredient in Chuanxiong Rhizoma that plays a protective role in the damage of optic nerve.Preliminary verification was conducted through in vitro experiments.The results suggest that Chuanxiong Rhizoma may interfere with autophagy and apoptosis,inhibit immune inflammation,as well as reduce oxidative stress in the treatment of glaucoma through the active components represented by CHR,so as to resist progressive optic nerve injury.Our study provides theoretical basis for the clinical use of Chinese herbal medicine or its extract in glaucoma,and also lays a solid foundation for the research of Chinese medicine in the field of optic nerve protection.

Exploring the scientific rationality of“Different dosage forms of the same prescription”base on Q-markers of pulvis and pill of Chuanxiong Chatiao Prescription

Objective:To predict and analyze the potential Q-markers of Chuanxiong Chatiao Prescription,and the pharmacokinetic properties of pulvis and pills in vivo were studied,which provided a basis for the rational evaluation of the phenomenon of“Different Dosage Forms of the Same Prescription”.Methods and Material:Q-markers analysis of Chuanxiong Chatiao Prescription based on the“Five Principles”(traceability and transmissibility,specificity,effectiveness,prescription compatibility and testability).The content determination method of Q-markers in Chuanxiong Chatiao Prescription was established by UPLC,and the content difference of Q-markers in the two dosage forms ware determined and compared.The Q-markers in rabbit plasma was determined by LC-MS/MS method,and the pharmacokinetic parameters of Q-markers in pulvis and pills were analyzed.Results:A total of 16 potential Q-markers from the“Five Principles”were used,nine components of tetramethylprazine,ferulic acid,glycyrrhizin,glycyrrhizic acid,luteolin,cimicifugoside,senkyunolideⅠ,isoimperatorin,nodakenin were identified as Q-markers of Chuanxiong Chatiao Presciption.The content of tetramethylprazine and other components in the pulvis form was found to be significantly higher than that in the pills,while the content of senkyunolideⅠwas lower than that in the pills,which may be related to the preparation process of the dosage form and the physicochemical properties of the components.Compared with pulvis,the Tmax and t_(1/2)of ferulic acid and other components in pills were significantly prolonged.To a certain extent,it can explain the classical theory of traditional Chinese medicine“Components in pulvis release quickly and take effect in fast-acting manner,while in pills release slowly and take effect in slow-acting”.Meanwhile,the Cmax and AUC0-t of tetramethylprazine and other components in pills were higher than those in pulvis,which showed unexpected pharmacokinetic characteristics,indicating the complexity of compounding and the importance of dosage form design.Conclusions:A method for the determination of Q-markers content was established by UPLC,which provide reference for the quality control of Chuanxiong Chatiao Prescription.In vivo studies have found the pharmacokinetic parameters indicate the absorption and distribution characteristics of pulvis and pills.However,it is also found that the release behavior of different components not only affected by the dosage form but also closely related to their own physical and chemical properties.

川芎挥发油治疗心绞痛的网络药理学研究及实验验证

目的 基于网络药理学研究川芎挥发油抗心绞痛的作用机制,并用动物实验加以验证。方法 通过水蒸气蒸馏法、气相色谱-质谱联用仪(gas chromatography-mass spectrometry, GC-MS)、口服利用度(oral bioavailability, OB)筛出挥发油成分;使用Pubchem、SwissTarget、DisGeNET、DrugBank数据库和R语言分别获取挥发油成分的靶点、心绞痛的靶点和交集靶点;蛋白质-蛋白质相互作用通过String数据库完成;使用R语言的ClusterProfiler包对交集靶点进行基因本体分析(gene ontology, GO)和京都基因与基因组百科全书分析(Kyoto Encyclopedia of Genes and Genomes, KEGG)富集分析,并通过Cytoscape构建中药-成分-靶点-通路网络;借助AutoDock vina 1.2.3、Pymol 3.0、Discovery Studio 2016软件进行分子对接,分析关键靶点和主要挥发油成分的亲和力。最后,通过动物实验进一步验证川芎挥发油对心绞痛的治疗效果。结果 网络药理学研究共得到10个挥发油成分和22个关键靶点,这些靶点与神经递质、突触膜上的受体、物质代谢等生物过程以及神经活性配体-受体相互作用、视黄醇代谢、药物代谢-细胞色素P450等通路密切相关;分子对接结果表明3-butylidenephthalide、Alpha-selinene、Trans-ligustilide等挥发油成分与多个关键靶点结合发挥治疗作用。动物实验表明川芎挥发油通过调节射血分数(ejection fraction, EF)、短轴缩短率(fractional shortening, FS)、左室收缩末内径(left ventricular internal diameter in systole, LVIDs)和每搏输出量(stroke volume, SV)水平以及乳酸脱氢酶(lactate dehydrogenase, LDH)、肌酸激酶(creatine kinase, CK)、天冬氨酸转氨酶(aspartate aminotransferase, AST)活性,促进受损心肌细胞中ADRA1A、CHRM5蛋白的表达,改善心肌纤维状态,减小细胞间隙,减少炎性细胞浸润表现出保护心肌的作用。结论 川芎挥发油具有有效保护受损心肌组织,治疗心绞痛的作用。

基于多元统计学分析^(60)Co-γ射线辐照对川芎挥发性成分的影响

川芎挥发性成分是其主要活性成分之一,也是评价川芎药用品质的重要指标。为探究^(60)Co-γ射线辐照对川芎挥发性成分的影响,本研究以不同吸收剂量处理川芎样品,采用电子鼻结合顶空固相微萃取-气相色谱-质谱技术(HS-SPME-GC-MS),通过主成分分析、线性判别分析、正交偏最小二乘法和聚类分析的多元统计学方法分析数据,以期了解川芎辐照前后挥发性成分的变化。电子鼻与GC-MS结果都表明:川芎气味主要由烷烃类、醇类、醛类、酮类贡献,10 kGy剂量以下的辐照处理对川芎挥发性气味成分影响不大;川芎经0 kGy(对照组)、3 kGy、7 kGy、10 kGy剂量辐照处理后,共鉴定出60种化合物,其中烃类23种,醇类12种,脂类10种,醛酮类4种,其他类11种;筛选到21种辐照前后的差异特征性指标物,它们在川芎挥发性成分中只占约12.92%,不是川芎的主要挥发性成分。经10 kGy剂量以下的辐照处理后,川芎中没有新生成的物质被鉴定出来。研究结果提示经10 kGy以下剂量处理川芎不会影响其主要挥发性成分。

基于GEO数据库联合网络药理学研究川芎-赤芍药对治疗动脉粥样硬化的药理过程及分子机制

目的:探讨活血化瘀中药药对川芎-赤芍拮抗动脉粥样硬化的潜在分子机制及药理过程。方法:使用R语言Limma包分析GEO数据库GSE43292数据集,对有表达差异的动脉粥样硬化基因进行筛选和提取。川芎-赤芍药对所含的化学活性组分及靶点通过中药系统药理学数据库与分析平台检索。合并差异基因和药物作用靶点以获得共同的靶点。利用在线分析工具STRING和Cytoscape构建药物和靶点的调控网络以及靶点间的蛋白质-蛋白质相互作用(PPI)网络。然后利用R语言注释靶点基因的基因本体(GO)功能,分析京都基因与基因组百科全书(KEGG)通路,再进行基因集富集分析(GSEA)以验证KEGG的通路和富集的情况,确定靶点基因的调控功能和参与基因调控功能的信号转导通道。结果:共筛选出动脉粥样硬化差异基因1244个,川芎-赤芍药对含36个生物活性成分,其中靶点为环加氧酶1、肾上腺素受体、过氧化物酶体增殖物激活受体-γ、激酶插入域受体、孕激素受体、基质金属蛋白酶9、CXC趋化因子配体8、蛋白激酶Cβ、白细胞介素6、CD14、二肽基肽酶-4、PIK3CG、肾上腺素受体α1B、微管相关蛋白2、尿激酶纤溶酶原激活剂和单胺氧化酶B。靶点在GO中主要富集在合成DNA过程的调控、DNA生物合成的过程、含胶原的细胞外基质、细胞外基质、蛋白酶结合、细胞迁移、血管形成过程、膜筏结构、转录的调节等与动脉粥样硬化炎症、脂肪代谢及血管生成相关的生物学注释。脂质与动脉粥样硬化通路和核因子κB信号通路与动脉粥样硬化关系最为密切,并且在KEGG信号通路和GSEA均显示出富集。结论:川芎-赤芍药对通过潜在的13个活性成分作用于可能的16个靶点调控相关信号转导通路,通过抗炎、调脂和保护血管等方式产生抗动脉粥样硬化的作用。

川芎祛瘀汤对关节镜下肩袖修补术后患者活动障碍和疼痛的影响

目的观察川芎祛瘀汤对关节镜下肩袖修补术后患者活动障碍和疼痛的影响。方法选取2020年1月—2023年1月期间安阳市中医院收治的90例肩袖损伤行关节镜下肩袖修补术患者,采用随机数字表法分为对照组和观察组,每组各45例。术后对照组予常规治疗及护理,观察组加用川芎祛瘀汤治疗。连续治疗4周。观察比较两组患者术前后疼痛、肩关节功能、肩关节活动情况、白细胞介素-6(Interleukin-6,IL-6)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)及血管内皮生长因子(Vascular endothelial growth factor,VEGF)表达。结果术后2、4周两组患者VAS评分均较术前降低,差异有统计学意义(P<0.05);且观察组术后2、4周VAS评分均明显低于对照组,差异有统计学意义(P<0.05)。术后4、8周两组患者UCLA评分及Constant-Murley评分均较术前明显下降,差异有统计学意义(P<0.05);且观察组术后4、8周UCLA评分及Constant-Murley评分均明显低于对照组,差异有统计学意义(P<0.05)。术后8周两组患者肩关节活动度均较术前明显升高,差异有统计学意义(P<0.05);且观察组肩关节活动度明显高于对照组,差异有统计学意义(P<0.05)。术后4周两组患者IL-6、TNF-α表达均较术前明显降低,VEGF表达均较治疗前明显升高,差异有统计学意义(P<0.05);且观察组IL-6、TNF-α含量均明显低于对照组,VEGF明显高于对照组,差异有统计学意义(P<0.05)。结论川芎祛瘀汤能有效缓解关节镜下肩袖修补术后患者疼痛,促进术后患者肩关节功能恢复,改善其关节活动,有效抑制炎症反应,促进血管再生,改善局部微循环。

川芎白芷药对对偏头痛小鼠行为学及血清NO、CGRP和IL-6含量的影响

目的 考察川芎白芷药对对偏头痛小鼠行为学及血清一氧化氮(nitric oxide, NO)、降钙素基因相关肽(calcitonin gene related peptide, CGRP)和白细胞介素-6(interleukin-6, IL-6)含量的影响,探讨川芎白芷药对对偏头痛的作用机制。方法 选取广西医科大学第二附属医院购买的56只SPF级雄性C57BL/6小鼠为研究对象,选取川芎白芷药对的指标性成分阿魏酸、欧前胡素为治疗药物,按处理方法不同将健康雄性小鼠随机分为7组,即空白组、模型组、阳性对照组、阿魏酸组、欧前胡素组、阿魏酸+欧前胡素低剂量组和阿魏酸+欧前胡素高剂量组,每组8只。采用硝酸甘油腹腔注射的方法制作偏头痛小鼠模型。造模成功后,对各组小鼠进行行为学考察,并检测血清中的NO、CGRP和IL-6的含量。结果 与对照组偏头次数(9.25±2.77)次比较,0~30 min内模型组小鼠挠头次数(26.25±4.60)次增加,阳性对照组(13.25±3.23)次,欧前胡素组(19.13±3.89)次,阿魏酸组(18.75±2.77)次,阿魏酸+欧前胡素低剂量组(16.88±2.52)次,阿魏酸+欧前胡素高剂量组(12.38±2.18)次,组间比较,差异有统计学意义(F=23.89,P<0.01)。模型组小鼠血清中NO、CGRP和IL-6的含量升高,差异有统计学意义(P均<0.05)。给予阿魏酸、欧前胡素或低、高剂量的阿魏酸+欧前胡素联合治疗后均能减少受试小鼠的挠头次数,并降低小鼠血清中NO、CGRP和IL-6的含量,其中阿魏酸+欧前胡素高剂量组的降幅最大,差异有统计学意义(P均<0.05)。结论 阿魏酸配伍欧前胡素后可有效改善偏头痛小鼠的症状,提示川芎白芷药对对偏头痛小鼠的作用机制可能与减少偏头痛小鼠血清NO、CGRP和IL-6水平含量有关。

川芎挥发油化学成分、制剂及药理作用研究进展

川芎是我国常用传统中药,其中挥发油是主要药效成分,一直是川芎研究工作的重点。早期对川芎挥发性成分的研究主要集中在其中化学成分的定性鉴定及定量分析。近年来研究者对川芎挥发油的研究逐渐深入,开展了川芎不同部位挥发油的差异分析,比较了加工制备方法对挥发油成分的影响;研究人员除了对川芎挥发油镇痛镇静、保护神经细胞、改善血管功能等药理作用及其机制进行深入研究外,新的研究发现川芎挥发油还具有骨保护、抗肿瘤、抗抑郁等药理活性。该文对近年来围绕川芎挥发油性成分的最新研究进展进行汇总,总结不同部位、不同产地、不同加工提取方法挥发油成分种类和含量差异,并综述了川芎挥发油制剂及药理活性研究进展,以期为更好地开发利用川芎挥发油自然资源提供参考。

川芎的化学成分和药理作用研究进展

川芎来源于伞形科植物川芎的干燥根茎,具有活血行气、祛风止痛的功效。川芎中主要含有苯酞、生物碱、酚酸、多糖等化学成分。药理研究表明,川芎对心脑血管系统、神经系统、呼吸系统等均具有一定的药理活性,主要表现为抗脑缺血、抗血栓、镇痛、抗炎、抗氧化、抗哮喘等药理作用。本文对川芎的化学成分及药理作用进行系统整理和归纳,以期为临床应用和资源开发提供参考。

基于全谱非靶向代谢组学技术的川芎不同部位代谢物深度解析

为了深度解析川芎中不同部位代谢物特征,本研究采用高效液相色谱法(HPLC)对川芎的根茎、茎、叶中的活性成分进行含量测定,并整合超高效液相色谱-质谱联用仪(UPLC-MS)和气相色谱-质谱联用仪(GC-MS)的全谱非靶向代谢组学技术,对川芎的根茎、茎和叶的挥发性成分、非挥发性成分进行全面的定性和定量分析。结果表明,川芎根茎中活性成分含量高于茎、叶。川芎的全谱非靶向代谢组学共检出2891个代谢物,总丰度为茎>叶>根茎,各部位化学成分种类相同、含量差异较大,包括氨基酸及其衍生物、萜类、酚酸类等32类化合物。其中LC-MS检出1726个代谢物,GC-MS检出1216个代谢物,两个平台共同检出51个代谢物。对川芎不同部位的差异代谢物进行分析,根茎与茎、根茎与叶、茎与叶中筛选到差异代谢物1683、2054和1844个,差异代谢物总丰度为根茎≈茎>叶。根茎中显著富集含氮化合物、酚类、其他类(糖类、内酯类),茎中的醇、胺类、醚类高度富集,叶中的萜类、酮类、黄酮类高度富集。川芎中大多数活性成分如川芎嗪、阿魏酸、藁本内酯等,呈现根茎>茎>叶的趋势,但茎和叶中也含有含量较高的阿魏酸、欧当归内酯A等重要活性成分,具有较高利用价值。通过KEGG富集分析结果推测差异代谢物可能与次生代谢产物的生物合成等途径相关。本研究深度解析了川芎不同部位的成分积累规律,为川芎资源的合理利用提供了科学依据。

基于人脐静脉内皮细胞的OGD/R模型研究当归-川芎药对中7个活性成分对VEGF-PI3K-AKT/NF-κB信号通路的影响

目的探讨当归-川芎药对中7个活性成分(绿原酸、阿魏酸、咖啡酸、丁烯基苯酞、洋川芎内酯H、洋川芎内酯A、藁本内酯)对VEGF-PI3K-AKT/NF-κB信号通路关键蛋白及其上下游血管活性物质、黏附因子和炎症因子的调控作用。方法构建人脐静脉内皮细胞(HUVEC)的氧糖剥夺/复氧(OGD/R)模型,采用细胞增殖试剂盒(CCK-8法)检测细胞活力,探索7个成分的最佳造模时间;通过检测乳酸脱氢酶(LDH)释放探索最佳给药浓度;采用酶联免疫吸附法(ELISA)检测7个成分对VEGF、VCAM-1、PAI-1、NF-κB、IL-1和IL-6表达的影响;采用逆转录-聚合酶链式反应(RT-PCR)法检测7个成分对VEGF-PI3K-AKT/NF-κB信号通路关键蛋白mRNA表达的影响。结果HUVEC氧糖剥夺6 h再复氧为最佳造模时间。高剂量绿原酸组、阿魏酸组、洋川芎内酯H组,低、中剂量丁烯基苯酞组,中、高剂量洋川芎内酯A、藁本内酯组显著降低LDH漏出率(P<0.05,P<0.01);绿原酸、阿魏酸、洋川芎内酯H部分剂量组细胞中VEGF、ICAM-1、VCAM-1的表达显著降低,绿原酸、阿魏酸、洋川芎内酯H、藁本内酯部分剂量组细胞NF-κB的表达显著下降,绿原酸、咖啡酸、丁烯基苯酞、洋川芎内酯H、洋川芎内酯A部分剂量组细胞中IL-6的表达显著增加,绿原酸、阿魏酸、洋川芎内酯A部分剂量组细胞IL-1表达显著减少,阿魏酸、洋川芎内酯H部分剂量组细胞中PAI-1的表达量显著下降(P<0.05,P<0.01);绿原酸、阿魏酸、咖啡酸、丁烯基苯酞和洋川芎内酯A部分剂量组细胞中ERK、VEGF、NF-κB、VEGFR2和MMP9的mRNA相对表达量显著下调,洋川芎内酯H和洋川芎内酯A部分剂量组细胞中AKT的mRNA相对表达量均显著上调(P<0.05,P<0.01)。结论当归-川芎中的药效成分可能通过抑制黏附因子、炎症因子和VEGF-PI3K-AKT/NF-κB信号通路关键蛋白mRNA的表达,从而发挥抗缺血性中风的作用。

川芎对肺癌干细胞原发性肿瘤相关黏附分子影响的实验研究

目的:探究川芎对肺癌干细胞样细胞荷瘤小鼠原发肿瘤的黏附作用机制。方法:通过无血清培养技术,培养肺癌细胞中的干细胞样细胞,并将其种植在小鼠脚垫。按照处理药物分为空白对照组(Con组)、川芎组(RC组)、顺铂组(DDP组)、川芎+顺铂组(RC+DDP组)并干预,3周后,在无菌环境中手术剥取肿瘤组织。通过Western blot实验技术和免疫组化技术,检测分析小鼠肿瘤细胞中相关黏附蛋白CD44v6、β1整合素的表达。结果:CD44v6在RC+DDP中的表达较DDP组、RC组及Con组减少,且差异具有统计学意义(均P<0.05)。β1整合素原发瘤组织在RC+DDP组中的表达低于Con组、RC组及RC+DDP组(均P<0.05)。结论:川芎和顺铂的联合应用,在一定程度上可减少肺癌干细胞样细胞原发肿瘤中黏附分子CD44v6、β1整合素的表达,从而抑制其黏附能力。

基于网络药理学研究川芎-赤芍治疗冠状动脉粥样硬化性心脏病心绞痛的作用机制

目的:通过网络药理学理念和技术研究川芎-赤芍药对治疗冠状动脉粥样硬化性心脏病(简称冠心病)心绞痛的主要活性成分、靶点及生物学过程和通路。方法:通过TCMSP数据库检索出川芎和赤芍的主要活性成分、靶点信息,再通过GeneCards、OMIM、TTD数据库获取冠心病心绞痛的相关靶点信息,继而得到川芎-赤芍与冠心病心绞痛的交集靶点,采用Cytoscape 3.10.0软件构建中药-成分-靶点-疾病靶点网络,将获取的交集靶点信息通过STRING数据库构建蛋白质-蛋白质相互作用网络,筛选出核心靶点。利用David数据库对交集靶点进行基因本体论(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。结果:从川芎和赤芍中共筛选得到36个主要活性成分,其中川芎7个、赤芍29个,相关作用靶点共166个,筛选去重后得到76个,冠心病心绞痛的相关作用靶点共1213个,川芎和赤芍与冠心病心绞痛交集靶点共35个,核心靶点18个。GO功能富集分析结果显示主要涉及对外源刺激的反应、凋亡过程的负调控、细胞外区域、质膜、酶绑定、相同蛋白结合等生物过程,KEGG通路富集分析结果显示主要涉及癌症通路、流体剪切力与动脉粥样硬化、卡波西肉瘤相关疱疹病毒感染、癌症中的蛋白聚糖类、化学致癌作用-受体活化、血脂与动脉粥样硬化等信号通路。结论:本研究初步揭示了川芎-赤芍药对治疗冠心病心绞痛疾病的主要活性成分、靶点、生物学过程和通路,为进一步研究其药理学作用及临床应用提供参考依据。

基于正交设计优化新鲜川芎中挥发油提取工艺

为优化川芎挥发油提取工艺,建立以新鲜川芎为原料提取制备挥发油的新方法,采用正交试验设计对乙醇回流提取工艺条件进行优化,并进一步考察优选回收浓缩、油水分离等关键工艺参数。以藁本内酯和洋川芎内酯A作为指标成分,应用HPLC一测多评法进行含量测定。以提取量、油收率、油含量为评价指标,最终确定川芎挥发油提取制备新方法为:鲜川芎切片,加8倍量70%乙醇回流提取2次(以干药材计,第一次需测定鲜川芎水分,调节乙醇浓度为70%),每次1.5 h,滤液回收浓缩至2.0~3.0 g生药/mL,静置过夜,弃去水层,上层乳化层加热至60℃离心,分取油层,余下乳化层再加氯化钠至饱和并加热至60℃离心,合并油层,即得川芎挥发油。新方法所得川芎挥发油收率约为3%,藁本内酯和洋川芎内酯A含量达70%。该研究所建立的新方法生产设备要求低、操作简便易行,可实现川芎挥发油的高效制备,且含量高、品质优,为川芎挥发油的工业化生产提供了新思路和新方法。

基于网络药理学与分子对接预测川芎-赤芍药对治疗动脉粥样硬化的作用机制

心血管疾病是全球范围内死亡率最高的一种疾病,其主要病理基础是动脉粥样硬化。为了探讨川芎-赤芍药对治疗动脉粥样硬化的潜在作用机制,利用网络药理学平台,挖掘了川芎-赤芍的有效成分及靶点并筛选核心成分;查找了动脉粥样硬化的靶点,然后与药物靶点取交集筛选候选靶点;构建候选靶点的蛋白互作网络得到核心靶点;对候选靶点进行GO功能富集分析与KEGG信号通路富集分析。最后将核心成分和核心靶点进行分子对接。结果从川芎-赤芍药对中筛选到包括核心成分β-谷甾醇、黄芩素、豆甾醇和杨梅酮在内的35个活性成分,获得川芎-赤芍药对治疗动脉粥样硬化的包括核心靶点AKT1、TNF、VEGFA、TP53在内的候选靶点58个,核心成分和核心靶点间均具有较好的结合能力,候选靶点主要参与了对氧水平下降的反应、对细菌来源分子的反应、对激素的反应等生物学过程,富集在癌症信号通路、脂质和动脉粥样硬化信号通路、化学致癌-受体激活通路、流体剪切力与动脉粥样硬化通路等相关通路上,以协同的形式通过调节细胞生长发育、降低血脂水平和控制促炎介质的分泌等对动脉粥样硬化发挥治疗作用,为川芎-赤芍药对的临床应用提供理论基础。

基于网络药理学及分子对接分析当归-川芎药对治疗银屑病的作用机制

目的:运用网络药理学方法和分子对接技术,预测当归-川芎药对治疗银屑病的潜在分子作用机制。方法:通过中药系统药理学数据库与分析平台筛选出当归和川芎的有效活性成分以及具体的功效靶点,通过检索GeneCards、人类孟德尔遗传综合数据库、DrugBank和DisGeNET数据库,获得银屑病相关靶点。利用Venn网页工具获取药物疾病共同靶点,将其输入Cytoscape 3.9.1软件绘制当归、川芎-共同靶点-银屑病网络图。运用STRING数据库及Cytoscape 3.9.1软件构建蛋白质-蛋白质相互作用网络,通过DAVID数据库进行基因本体(GO)、京都基因与基因组百科全书(KEGG)富集分析,用微生信绘制富集分析的气泡图。利用AutoDock软件对分子对接进行验证,并用PyMOL软件对分子对接进行可视化。结果:获取当归、川芎治疗银屑病的有效活性成分9个,潜在作用靶点28个,关键作用靶点为胱天蛋白酶3、环氧化酶2、雌激素受体1、转录因子AP-1和丝裂原激活的蛋白激酶14。主要涉及药物的反应信号通路、细胞内类固醇激素受体信号通路、磷脂酰肌醇3激酶-蛋白激酶B信号通路和白细胞介素17信号通路等。结论:当归-川芎药对通过多通路、多种有效活性成分和多种有效靶点实现对银屑病的治疗。