Exploration of the effective components and mechanism of Alpinia oxyphylla-Saposhnikovia Radix in the treatment of diabetic kidney disease based on network pharmacology

Objective:Network pharmacology was used to predict the main targets and related mechanism pathways of the key compounds and active ingredients of the Alpinia oxyphylla-Saposhnikovia Radix based on the therapeutic strategy of tonifying kidney and dispelling pathogenic wind for the treatment of diabetic kidney disease(DKD).Methods:The effective components of Alpinia oxyphylla-Saposhnikovia Radix were screened through the data platform of Traditional Chinese Medicine(TCM).After screening effective targets,the disease target gene database was used to obtain targets for DKD,and then gene enrichment analysis to predict relevant signalling pathways after obtaining the key target genes through Venn analysis.Results:There were 324 active components of Alpinia oxyphylla-Saposhnikovia Radix through the data platform of TCM.In addition,Venn analysis showed that there were 48 key target genes for the treatment of DKD.The biological pathways for the action of Alpinia oxyphylla-Saposhnikovia Radix in DKD were closely related to many signaling pathways including NF-kappaB,classical MAP kinase,JNK and P38 MAP kinase,and ERK5.Conclusions:Treatment of DKD with Alpinia oxyphylla-Saposhnikovia Radix based on the therapeutic strategy of tonifying kidney and dispelling pathogenic wind has the characteristics of multi-components,multi-targets and multi-pathways,which could provide new ideas and methods for further researches on the treatment of DKD with Alpinia oxyphylla-Saposhnikovia Radix.

Network pharmacology and verification experiment-based prediction of active components and potential targets of Alpiniae Oxyphyllae Fructus-Saposhnikoviae Radix(Yizhiren-Fangfeng)for treatment of diabetic kidney disease

Background:In this study,we analyzed the potential active components,related crucial targets and possible signaling pathway mechanisms of Alpiniae Oxyphyllae Fructus and Saposhnikoviae Radix(AOF-SR)herb pairs in the treatment of diabetic kidney disease(DKD)using network pharmacology and verification experiments.Methods:The active compounds and potential targets of AOF-SR were derived from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,The Encyclopedia of Traditional Chinese Medicine,and PubChem databases,and the potential therapeutic targets of DKD were derived from the OMIM,Drugbank,and DisGeNET databases.The“compounds-diseases-targets”network was constructed using Cytoscape 3.6.0.ClusterMaker functionality in Cytoscape is being used to screen important targets for AOF-SR treatment of DKD.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis of important targets were performed using DAVID database.In addition,according to the predicted results of network pharmacology,HK-2 cells were used to construct DKD model for verification experiment.HK-2 cells were divided into control group,high glucose(HG)group and AOF-SR(HG+AOF-SR)group to detect survival rate and expression of key proteins in NF-κB and PI3K/Akt signaling pathways.Results:A total of 38 compounds were selected from AOF-SR,of which 23 were Alpiniae Oxyphyllae Fructus and 15 were Saposhnikoviae Radix.Through enrichment analysis of 82 important targets,88 signaling pathways were identified;some of these pathways,such as the NF-κB,PI3K-Akt,IL-17,and JAK/STAT signaling pathways,regulate the pathological process of DKD.In verification experiment,the HK-2 cells survival rate was higher in the HG+AOF-SR group than in the HG group(P<0.05).Moreover,western blotting results showed that the expression levels of NF-κB,p-PI3K,and p-Akt in HG+AOF-SR group were significantly lower than those in HG group(P<0.05).Conclusion:Overall,this study revealed the active compounds,important targets and possible mechanisms of AOF-SR treatment for DKD,and conducted preliminary verification experiments on its correctness,provided novel insights into the treatment of DKD by AOF-SR.

Effects and its possible mechanism of Radix Saposhnikoviae on rat colonic smooth muscle in vitro

Objective： To determine the effect of different concentrations of Radix Saposhnikoviae （RS） on the contraction of smooth muscle strips and the Ca2＋ mobilization of cultured smooth muscle ceils of rat colon and its possible mechanism of action. Methods： Strips of rat colon longitudinal muscle were prepared and smooth muscle cells from rat colon were isolated and cultured. In the experiments, in vitro muscle strips were suspended in an organ bath and the contraction of the strips was recorded. In the cell- experiments, intracellular Ca2＋ was assessed using fluorescent intensity （FI） of smooth muscle cells loaded with Fluo-4/AM, measured with a laser scanning confocal microscope and related software. Results： In the in vitro experiment, RS （0.02, 0.2, 2, 20 g/L） inhibited contraction of muscle strips in a concentration-dependent manner, and this inhibition was significant for the three higher RS concentrations （P 〈 0.01） for both Peak （the maximal contraction amplitude） and Area （the area under curves）. Similarly, RS inhibited Ach-induced contraction. In these experiments the inhibition of the Peak values in the RS 2 and 20 g/L groups was significant （P 〈 0.01）, as was the inhibition of the Area values in all RS groups （P 〈 0.05）. Naloxone and propranolol did not significantly affect the inhibitory effect of RS on smooth muscle contractility, while phentolamine significantly reduced the inhibitory effect （P 〈 0.01）. In experiments using primary smooth muscle cell cultures in Ca2＋ - containing buffer, the post-treatment fluorescence of cells in the RS 0.2, 2 and 20 g/L groups differed significantly from pre-treatment values （P 〈 0.05）, and the percent inhibition of fluorescence in the RS 2 g/L and 20 g/L groups was significant （P 〈 0.01）. However, in Ca2＋-free buffer, FS had no significant effect on cell fluorescence. Conclusion： RS inhibited both the spontaneous and Ach-stimulated contraction of rat colonic smooth muscle strips. This RS effect appeared to involve α -adrenoceptors, but not β -adrenoceptors or opioid receptors. In cultured primary smooth muscle cells, RS reduced the mobilization of Ca2＋ from extracellular sources, but may had no effect on the release of Ca2＋ from sarcoplasmic reticulum and endoplasmic reticulum.

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.

基于颜色数字化结合熵权法分析防风色度值与内在成分的相关性

目的:基于颜色数字化结合熵权法,分析防风色度值与其内在成分的相关性。方法:采用分光测色仪测定防风药材粉末的色度值L^(*)、a^(*)、b^(*)及总色差值ΔE^(*),测定药材中醇溶性浸出物和水溶性浸出物含量,采用UHPLC法同时测定升麻素苷、升麻素、5-O-甲基维斯阿米醇苷、亥茅酚苷、亥茅酚、3′-O-乙酰基亥茅酚6个成分的含量,利用熵权法对内在成分进行综合得分,并进行相关性分析。结果:L^(*)值与水溶性浸出物和醇溶性浸出物含量呈显著正相关,与升麻素苷、升麻素、亥茅酚苷、亥茅酚、3′-O-乙酰基亥茅酚含量呈显著负相关;b^(*)值仅与亥茅酚苷含量呈显著正相关;ΔE^(*)与水溶性浸出物和醇溶性浸出物含量呈显著负相关,与升麻素、亥茅酚苷、亥茅酚、3′-O-乙酰基亥茅酚含量呈显著正相关。各指标综合得分与L^(*)呈显著负相关,与ΔE^(*)呈显著正相关。结论:防风粉末颜色越深,其质量越好,该研究丰富了防风药材质量评价方法,为防风药材“辨色论质”提供了依据。

国医大师禤国维治疗儿童特应性皮炎用药规律的挖掘

【目的】采用数据挖掘技术挖掘国医大师禤国维教授治疗儿童特应性皮炎的组方用药规律。【方法】收集禤国维教授门诊治疗儿童特应性皮炎的有效病例的病案资料,采用频数统计、关联规则分析、聚类分析等方法对录入的病案数据进行统计分析。【结果】共收录处方242首,涉及中药101味。常用药物为23味,高频药物(频数>100次)共16味,分别为甘草、防风、北沙参、紫苏叶、麦冬、徐长卿、布渣叶、白鲜皮、玄参、薏苡仁、蝉蜕、百合、生地黄、地肤子、茯神、芡实。药物的性味归经分析显示,药物大多偏甘寒,主要归脾、胃、肝经。关联规则分析得到常用药物组合24个,关联规则20条。聚类分析得到2个核心药物组合。【结论】禤国维治疗儿童特应性皮炎以“清、补、和”三法为主,“扶正御邪,平调阴阳”的用药原则贯穿治疗始终。

龙泽熊胆胶囊中水防风掺伪的检测方法研究

目的建立龙泽熊胆胶囊中水防风掺伪的检测方法。方法以水防风区别于防风的专属性成分Xanthalin建立薄层色谱鉴别法(TLC),对样品进行初筛。若供试品溶液色谱中出现水防风特征斑点,则采用高效液相色谱(HPLC)进行定性检查。色谱条件:SHIMADZU InertSustain~?C18色谱柱(250 mm×4.6 mm,5μm);以乙腈-水(58∶42)为流动相,等度洗脱;流速:1.0 mL·min^(-1);柱温:30℃;检测波长:324 nm;进样量:10μL。若供试品Xanthalin色谱峰面积大于对照品的峰面积,视为阳性检出,必要时可进一步采用超高效液相色谱串联三重四级杆质谱法(UPLC-MS/MS)进一步确证。结果建立的TLC法、HPLC法简便、快速,专属性、重复性、耐用性均符合《中国药典》要求。对市售107批龙泽熊胆胶囊样品进行检测,19批次样品检出Xanthalin,提示该制剂存在水防风掺伪投料生产的风险。结论该方法可实现水防风掺伪防风的定性鉴别,有助于完善该品种的质量控制。

李云英治疗鼻鼽用药规律的数据挖掘

【目的】采用数据挖掘方法探析李云英教授治疗鼻鼽的用药规律。【方法】收集李云英教授门诊辨治鼻鼽的有效病例的处方,采用R语言中的相关软件进行频次分析、关联规则分析、聚类分析。【结果】共纳入1682首中药处方,涉及单味中药173味。将使用频次居前30位者定义为高频中药,高频中药主要包括甘草、辛夷、防风、白芷、苍耳子、白术、白芍、地龙、细辛、蒺藜、茯苓、黄芩、党参、蜂房、路路通等。关联规则分析得到二阶有效关联规则10条,三阶有效关联规则42条,四阶有效关联规则64条。聚类分析得到5类药物组合。【结论】李云英教授治疗鼻鼽的核心药物配伍为苍耳子、辛夷、白芷、白术、防风、白芍;常用方剂有二陈汤、苍耳子散、辛夷散、玉屏风散、四君子汤、桂枝汤、痛泻要方;总以祛风散邪、宣利鼻窍、健脾益肺为法。其用药规律可为中医临床治疗鼻鼽提供借鉴。

范瑞强辨治荨麻疹经验

荨麻疹是一种临床常见的变态反应性皮肤疾病。范瑞强教授认为,荨麻疹主要病机为禀赋不耐,风邪是主要致病因素,岭南地区患者易夹湿夹热。荨麻疹的临床常见证型为风湿热证、脾虚湿蕴证、营卫不和证等。荨麻疹的分期施治重在辨虚实,急性荨麻疹以实证居多,慢性荨麻疹以虚证居多,或虚实夹杂。治疗实证者以清热除湿、祛风止痒为法,处方以搜风利湿方加减;虚实夹杂者需标本兼治,健脾化湿止痒,处方以多皮饮加减;虚证者宜调和营卫,祛风止痒,处方以玉屏风散合桂枝汤加减。范瑞强教授临证治疗荨麻疹的高频中药主要为防风、徐长卿、荆芥、生甘草、合欢皮、茵陈、白芍、连翘、蒺藜、鱼腥草、生地黄、白鲜皮、白术、茯苓、黄芪。范瑞强教授临证治疗荨麻疹用药灵活,以减少复发、提高患者生活质量为主要目的。

龙泽熊胆胶囊质量标准提升研究

目的提升龙泽熊胆胶囊的质量标准。方法采用薄层色谱(TLC)法对防风药材中的升麻素和5-O-甲基维斯阿米醇进行定性鉴别,并进行耐用性考察。采用高效液相色谱法同时测定制剂中上述2种成分的含量,色谱柱为Wondasil C18柱(200 mm×4.6 mm,5μm),流动相为乙腈-水(梯度洗脱),流速为0.8 mL/min,检测波长为254 nm,柱温为30℃,进样量为20μL。结果TLC图斑点清晰、分离度好,且阴性对照无干扰。升麻素和5-O-甲基维斯阿米醇进样量分别在0.051~1.020μg、0.025~0.499μg范围内与峰面积线性关系良好(R^(2)=1.000);检测限均为0.1μg/mL,定量限均为0.3μg/mL;精密度、稳定性、重复性试验结果的RSD均小于2.0%;平均加样回收率分别为96.20%和91.40%,RSD分别为1.09%和1.89%(n=6)。结论所建立的方法专属性强,重复性、耐用性好,结果稳定,可为龙泽熊胆胶囊的质量控制提供参考。

中药防风与前胡的比较研究概况

防风和前胡为常用中药,因药材性状相似,鉴定较为困难,经常出现混用或错用的现象。两者均来源于伞形科植物的干燥根,化学成分相似,临床上均可用于治疗感冒,但两种药材之间存在明显的差异。防风始载于《神农本草经》,为祛风要药,主要化学成分为色原酮类化合物;前胡始载于《名医别录》,为化痰止咳平喘药,主要化学成分为香豆素类化合物。通过对防风与前胡的异同点进行梳理总结,以期为两者的快速、准确鉴定和选用提供新的研究思路和解决方法,同时保障临床用药的安全性。

基于色度分析原理的防风有效成分含量与颜色值相关性研究

目的研究防风有效成分含量与颜色值之间的相关性。方法尝试从传统中医药经验角度出发,进入色度计对野生防风与栽培防风粉末颜色进行测量,同时结合药材内部物质成分含量测定结果,对二者间相关性进行分析研究。结果升麻素苷含量与L*,a*,b*,Eab的相关系数为-0.319,-0.47,-0.448,-0.351,其相应的P值分别为0.062,0.004,0.007,0.039;5-O-甲基维斯阿米醇苷含量与L*,a*,b*,Eab的相关系数为0.181,0.009,0.105,0.159,相应的P值分别为0.297,0.957,0.549,0.363。结论防风中升麻素苷含量与a*,b*,Eab值存在显著的负相关关系,5-O-甲基维斯阿米醇苷含量与a,b,Eab值相关性不显著。防风颜色值主要与防风中升麻素苷含量有关。

采用体外肠道菌群实验研究防风对黄芪中毛蕊异黄酮葡萄糖苷代谢的影响

目的研究防风对黄芪中主要活性成分毛蕊异黄酮葡萄糖苷在大鼠肠道菌群中代谢的影响,从代谢的角度探讨黄芪-防风药对配伍应用的科学内涵。方法通过肠道菌群实验比较不同时间点黄芪组和黄芪-防风组以及毛蕊异黄酮葡萄糖苷单体组和毛蕊异黄酮葡萄糖苷-防风组中毛蕊异黄酮葡萄糖苷、毛蕊异黄酮的浓度。结果与黄芪组相比,不同时间点黄芪-防风组毛蕊异黄酮葡萄糖苷的浓度均增高,毛蕊异黄酮的浓度均降低。与毛蕊异黄酮葡萄糖组相比,不同时间点毛蕊异黄酮葡萄糖苷-防风组毛蕊异黄酮葡萄糖苷的浓度均增高,毛蕊异黄酮的浓度均降低。结论防风能抑制黄芪中毛蕊异黄酮葡萄糖苷转化为毛蕊异黄酮的水解反应,减缓其代谢。

常用治疗急性湿疹的中药抗Ⅳ型超敏反应的实验研究

目的研究常用治疗急性湿疹的单味中药(防风、甘草、地肤子、薏苡仁、金银花)抗Ⅳ型超敏反应作用,探讨中药作用机理。方法使用2,4-二硝基氯苯(DNCB)诱导建立小鼠变应性接触性皮炎(ACD)的模型,观察中药对小鼠耳的肿胀度、外周血白细胞数的影响,采用酶联免疫法(ELISA)检测各组小鼠血清中的白介素-4(IL-4)、γ-干扰素(IFN-γ)及可溶性白介素-2受体(sIL-2R)的水平。结果防风、甘草、地肤子、薏苡仁均可抑制DNCB引起的小鼠耳部超敏反应性肿胀所造成的重量改变(P<0.05)。5种中药均可降低DNCB引起的小鼠耳部肿胀所造成的厚度改变,防风作用较明显。5种中药均可降低DNCB引起的白细胞总数升高,薏苡仁的作用最弱。防风、地肤子、薏苡仁均可使ACD小鼠模型升高的IL-4水平下降,且与DNCB模型组相比较有差异显著性(P<0.05)。甘草、金银花不能使IL-4下降。5味中药均显著提高下降的IFN-γ、sIL-2R的含量,防风、地肤子能明显提高IFN-γ的含量。结论中药抗Ⅳ型超敏反应的作用机理与抑制白细胞总数、调节细胞因子及受体有关。

防风与刺蒺藜的药理实验研究

目的 :研究防风与刺蒺藜单用及合用的药理作用。方法 :运用右旋糖酐及组胺造成搔痒模型 ,组胺引起毛细血管通透性增高 ,二甲基亚砜引起接触性荨麻疹 ,观察药物作用。结果 :防风、刺蒺藜、防刺合煎剂对外源性组胺和右旋糖酐诱导内源性组胺动物搔痒模型均有显著的止痒作用 ,能明显抑制组胺引起的毛细血管通透性增高 ,能不同程度地抑制二甲基亚砜所致的豚鼠耳肿胀。结论 :防风、刺蒺藜、防刺合煎剂有抗过敏作用 。

防风有效部位的药理作用研究

目的:观察防风有效部位药理作用。方法:采用醋酸致炎法,抗凝血和血液流变学方法研究相关药理作用。结果:防风有效部位大剂量可使醋酸致炎渗出液减少,降低血浆黏度。防风有效部位中剂量和大剂量能延长凝血酶原时间和抑制由ADP诱导的血小板聚集。结论:防风有效部位有明显的抗炎、降低血浆黏度、延长凝血酶原时间和抗血小板聚集作用。

RRLC-TOF/MS鉴别防风血浆、尿液中成分及代谢产物

目的应用高分离度快速液相-飞行时间质谱(RRLC-TOF/MS)联用技术鉴别防风入血成分及尿液中的原型成分及代谢产物,以探索防风可能的药效成分及体内代谢过程。方法采用Angilent Zorbax Extend-C18柱(250mm×4.6mm,5μmid),甲醇-水流动相体系梯度洗脱,在飞行时间质谱正离子模式下扫描,利用TOF/MS得到的准确相对分子质量,对照化学成分数据库,对血浆、尿液中的防风成分及代谢物进行鉴别。结果鉴别出血浆中6个防风原型成分:蔗糖、升麻苷、升麻素、紫花前胡苷元、5-O-甲基维斯阿米醇苷和3'-O-i-丁酰亥茅酚;尿液中8个原型成分:升麻苷、divaricatacid、升麻素、5-O-甲基维斯阿米醇苷、(3S)-2,2-二甲基-3,5-二羟基-8-羟甲基-3,4-二氢-2H,6H-苯并-[1,2-b:5,4-b']双吡喃-6-酮、5-O-甲基维斯阿米醇、亥茅酚苷和汉黄芩素;及2个代谢产物:升麻素的葡萄糖醛酸苷和升麻素的同分异构体。结论该方法可靠、有效,可成功用于防风体内成分鉴别,并为进一步的药效实验提供参考和依据。

HS-SPME-GC-MS联用分析羌活、防风药对配伍前后挥发性组分变化规律

目的比较羌活、防风药对配伍前后挥发性组分变化。方法采用顶空固相微萃取法(HS-SPME)与气相色谱-质谱联用(GC-MS)法,单因素考察并确定最佳萃取条件,测定羌活、防风以及羌活、防风药对的挥发性化学成分,通过面积归一法确定各成分含量。结果最佳萃取条件为取样量0.5g,萃取温度80℃,萃取时间15min,解析时间3min。测出92个羌活挥发性成分,71个防风挥发性成分,150个药对挥发性成分,分别占挥发性成分总面积的70.90%、88.52%和81.48%。药对中新增43种新的成分,单味药中有34种成分在药对中未能检测出,44种共有成分含量减少,10种共有成分含量增加。结论羌活、防风药对的挥发性成分大部分来自于羌活,羌活、防风配伍后有新的活性成分产生,该药对配伍可能具有协同增效作用。

防风与刺蒺藜抗Ⅰ型变态反应的实验研究

目的:探讨防风、刺蒺藜单用及二药合用的抗Ⅰ型变态反应作用。方法:采用小鼠同种PCA反应模型、大鼠颅骨骨膜肥大细胞脱颗粒模型及Schu lz-Dale反应模型进行观察。结果:防风、刺蒺藜、防刺合煎剂均可明显抑制小鼠同种PCA,大鼠颅骨骨膜肥大细胞脱颗粒及Schu lz-Dale反应。结论:防风、刺蔟藜、防刺合煎剂均有抗Ⅰ型变态反应作用,防风与刺蔟藜合用的效应较单用为相加。

痛泻要方中配伍“风药”对PI-IBS模型大鼠PAR_2 mRNA表达及炎症介质的影响

目的探讨痛泻要方中配伍风药(防风)对感染后肠易激综合征(PI-IBS)大鼠模型PAR2mRNA表达和炎症介质的影响。方法采用福氏痢疾杆菌灌胃法复制PI-IBS大鼠模型,评价痛泻要方原方、痛泻要方无防风方、防风的疗效及对结肠黏膜PAR2mRNA基因表达和炎症介质SP、TNF-α、IL-6的影响。结果痛泻要方原方、痛泻要方无防风方、防风可升高感染后IBS大鼠的内脏痛觉敏感压力阈值,减少大鼠排稀便次数(P<0.05或P<0.01);痛泻要方原方及防风组显著降低结肠黏膜SP、IL-6、TNF-α含量并抑制结肠黏膜PAR2mRNA基因表达,而痛泻要方无防风方可降低结肠黏膜SP的含量(P<0.05),但对结肠黏膜IL-6、TNF-α和PAR2mRNA基因表达无显著影响(P>0.05)。结论痛泻要方中配伍风药(防风)可增强方中其他药物的止痛止泻效应,增强对PAR2基因表达和炎症介质的抑制作用,防风在方中发挥了不可替代的佐助作用。