Optimization of Extraction Process of Fagopyri Dibotryis Rhizoma by Orthogonal Experiment

[Objectives]To optimize the water extraction process of Fagopyri Dibotryis Rhizoma.[Methods]The entropy weight method was used to determine the weight of epicatechin extraction rate and dry extract rate and calculate the comprehensive score.The water extraction process of Fagopyri Dibotryis Rhizoma was optimized by orthogonal design with the comprehensive score as the indicator and the amount of water,extraction time and extraction times as the factors.[Results]The optimum extraction process of Fagopyri Dibotryis Rhizoma was as follows:adding 10 times of water,extracting 3 times,and extracting for 60 min each time.[Conclusions]The optimized extraction process is stable and feasible,and can be used for the extraction of Fagopyri Dibotryis Rhizoma.

Fingerprint Study of Polygonati Rhizoma with Steaming and Exposing to the Sun Alternatively for Different Times

[Objectives]To explore the influence of different times of steaming and exposing to the sun on the fingerprint of Polygonati Rhizoma by studying the HPLC fingerprint of Polygonati Rhizoma processed products with different times of steaming and exposing to the sun,and to provide a basis for the determination of the best processing technology of Polygonati Rhizoma.[Methods]SETSAIL II AQ-C 18(5μm×250 mm×4.6 mm)was used as the column,the column temperature was 30℃,pure water(A)and acetonitrile(B)were eluted gradually,0-10 min,B(5%-10%),10-30 min,B(10%-35%),30-40 min,B(35%-60%),40-45 min,B(60%-100%),flow rate 1 mL/min,absorption wavelength 200 nm.[Results]The relative retained peak area RSDs of the common peaks in the precision,reproducibility and stability tests were all less than 5%.There were 17 common peaks in the fingerprint of nine batches of samples,and the retention time of Peak 2 was basically the same as that of the reference peak of 5-HMF.Peak 4 mainly existed in the chromatogram of Sample 3 to Sample 5,peaks 5 and 11 mainly existed after Sample 3,peaks 9,14 and 16 mainly existed after Sample 6,and peaks 12 and 17 mainly existed after Sample 4.[Conclusions]A total of 17 common peaks were obtained,and the Peak 2 was the designated peak,and the chemical components of each processed product were different.

Mechanism of Polygoni Cuspidati Rhizoma in treating atherosclerosis based on network pharmacology and molecular docking

Background:The incidence and prevalence of atherosclerosis(AS)is increasing every year and has becoming a major health issue of global concern.Polygoni Cuspidati Rhizoma(PCR)is a Chinese herb that is widely used clinically for the treating of AS.However,its pertinent targets and probable mechanisms,still need to be completely explored.Methods:Active compounds and targets for PCR and AS targets were screened using public databases.A“drug-component-disease target”network map was created and analyzed after using the Venn online tool to identify common targets and Cytoscape software to screen drug-disease core targets.Critical targets pathway enrichment analyses are conducted using the Metascape database.Using AutoDock Vina and Pymol software,docking validation and visualization of active components and core targets were carried out.Results:PCR was obtained for ten compounds with 105 AS-related targets.Rhein,quercetin,beta-sitosterol,and luteolin may be drug candidates,and the genes for AKT1,TNF,IL-6,EGFR,TP53,IL-1,RELA,and VEGFA are potential therapeutic targets,according to network analysis.PCR might modulate the AGE/RAGE,PI3K/Akt,IL-17 and NF-ᴋB signaling pathways against the development of AS.Molecular docking indicated that quercetin has high affinity for AKT1 and TNF gene targets.Conclusion:This study provides rare information and scientific basis for further exploration of PC in the treatment of AS.

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.

Exploring the mechanism of active ingredients of Smilacis Glabrae Rhizoma in treating migraine headache through network pharmacology and animal experiments

Background:To explore the potential mechanism of action of the active ingredients of Smilacis Glabrae Rhizoma(SGR)in the treatment of migraine using network pharmacology and in vivo experiments.Methods:Through the search of Traditional Chinese Medicines Systems Pharmacology Database and Analysis Platform,Genecards,Drugbank and other databases,we obtained active ingredients,targets of SGR and related disease targets of migraine,and took the intersection for protein-protein interactions analysis.After constructing the network diagram,network topology analysis was performed to derive the core targets and key active ingredients,and Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed.Finally,molecular docking was performed and validated by in vivo experiments.In vivo experiments,18 male BALB/c mice were selected,and the SGR group was fed with SGR drinking tablet concentrate,and nitroglycerin injection was used to construct a mouse model of migraine.Enzyme-linked immunosorbent assay test was used to detect the levels of TNF-α,IL-1β,IL-6,and AKT1 in plasma.Results:The results showed that the core targets of SGR for the treatment of migraine were TNF-α,IL-1β,IL-6,and AKT1.These core targets and key active ingredients had better binding ability.Compared with the blank group,the number of head scratching in the model group increased.Compared with the model group,there was a significant reduction of the number of head scratching in the SGR group.In comparison with the blank group,the protein level in the plasma in the model group was markedly higher.Compared with the model group,the protein level in the SGR group was significantly lower.Conclusion:SGR has the characteristics of improving migraine based on multi-targets,multi-components and multi-pathways,and the mechanism of action may be related to the inhibition of the release of inflammatory factors,neuron protection,and interference with apoptosis and other processes.

Qualitative and Quantitative Analysis of Rhizoma of Polygonum cuspidatum

Aim To establish reliable methods for evaluating the quality of rhizoma of Polygonum cuspidatum( Huzhang in Chinese). Methods TLC and HPLC were employed for the chemical identification and content determination,respectively. Results A qualitative TLC method and a quantitative HPLC method with piceid as the reference substance were established, respectively. With piceid as the reference substance and ethyl acetate-methanol-formic acid-water ( 19:3:0.5:1) as the mobile phase, a TLC method for the identification of Huzhang from the commonly used crude drugs of the same family was also set up. Conclusion The established TLC method can reasonably appraise the quality of the drug and easily distinguish Huzhang from the other commonly used crude drugs of the same family. The HPLC method for determining piceid is simple, reproducible, accurate, and feasible.

Studies on the Basic Principles for the Processing of Rhizoma Cibotii Part Ⅰ Influence of Rhizoma Cibotii and Its Processed Samples on Thrombin Induced Rabbit Platelet Aggregation

比较研究了狗脊及其不同炮制品对凝血酶诱导的兔血小板聚集作用的影响 ,各炮制品均有抑制血小板聚集作用 ,抗血小板聚集作用砂烫品 >盐制品 >酒蒸品 >单蒸品 >

转录组测序分析黄连总碱抗脑缺血的作用机制

背景:黄连清热燥湿、泻火解毒,黄连及其成分对脑缺血有显著的保护作用,基于网络药理学和转录组测序探究黄连总碱抗脑缺血的作用机制。目的:在明确黄连总碱对脑缺血大鼠保护作用的基础上,基于网络药理学和转录组测序技术探讨黄连总碱干预脑缺血的作用机制。方法:将SD大鼠随机分为假手术组、缺血再灌注组、阳性药物组和黄连总碱组,后3组采用改良线栓法制备大脑中动脉阻塞的缺血再灌注模型,假手术组不插入线栓,其余手术操作相同。通过TTC染色、Longa 5神经功能缺损评分、苏木精-伊红染色、尼氏染色评价黄连总碱对脑缺血再灌注模型大鼠的脑保护作用。对假手术组、缺血再灌注组、黄连总碱组大鼠脑组织进行转录组测序,通过差异表达基因筛选、基因本体论分析、京都基因与基因组百科全书分析以及转录组学与网络药理学关联分析,阐明黄连总碱干预脑缺血的作用特点,最后通过ELISA检测及免疫荧光染色对黄连总碱干预脑缺血的关键靶点进行验证。结果与结论:①黄连总碱可降低缺血再灌注模型大鼠Longa 5神经功能缺损评分及脑梗死面积,增加神经元、尼氏小体数目;②黄连总碱治疗后差异表达基因的基因本体论功能富集分析包括炎症反应、分裂原活化蛋白激酶级联的正调控等生物学过程;京都基因与基因组百科全书通路富集分析主要涉及白细胞介素17信号通路、神经活性配体-受体相互作用、环磷腺苷信号通路等通路;③转录组分析发现黄连总碱主要调控的基因有前列腺素内过氧化物合酶2、脑源性神经营养因子、瞬间受体电位A1等;④采用网络药理学分析发现,黄连总碱中9个成分可能通过87个成分靶点关联到过氧化物合酶2、脑源性神经营养因子及瞬间受体电位A1而发挥作用;⑤ELISA检测及免疫荧光染色结果进一步证实,黄连总碱调控脑缺血再灌注模型大鼠过氧化物合酶2、脑源性神经营养因子和瞬间受体电位A1的表达;⑥提示黄连总碱能明显改善脑缺血再灌注模型大鼠损伤,可能通过调控过氧化物合酶2、脑源性神经营养因子和瞬间受体电位A1而发挥作用。

Study on Rhizoma Chuanxiong based on capillary electrophoresis with amperometric detection

A high-performance capillary electrophoresis with amperometric detection(CE-AD) method has been developed for the analysis of seven bioactive ingredients,namely ferulic acid(FA),vanillin,vanillic acid,p-hydroxybenzoic acid,caffeic acid,gallic acid and protocatechuic acid,in Rhizoma Chuanxiong.The effects of several factors such as the acidity and concentration of running buffer,the separation voltage,the applied potential to working electrode and the injection time were investigated.Under the optimum condit...

Effects of Gastrodiae rhizoma on proliferation and differentiation of human embryonic neural stem cells

Objective:To investigate the effects of Gastrodiae rhizoma,a dried root of Gastrodia elata Blume,on proliferation and differentiation of human NSCs derived from embryonic stem cells.Methods:A 70%ethanol extract of Gastrodiae rhizoma(EEGR) was estimated with4-hydroxybenzyl alcohol as a representative constituent by HPLC.Results:MTT assay showed that the treatment with EEGR increased the viability of NSCs in growth media.Compared to contro1,EEGR increased the number of dendrites and denritic spines extended from a differentiated NSC.Whereas EEGR decreased the mRNA expression of Nestin,it increased that of Tuj1 and MAP2 in NSCs grown in differentiation media.Immunocytochemical analysis using confocal microscopy also revealed the increased expression of MAP2 in dendrites of EEGR-treated NSCs.Furthermore,EEGR decreased mRNA expression of Sox2 in NSCs grown even in growth media.Conclusions:In conclusion,our study demonstrates for the first time that EEGR induced proliferation and neuronal differentiation of NSCs,suggesting its potential benefits on NSC-based therapies and neuroregeneration in various neurodegenerative diseases and brain Injuries.

Effects of Rhizoma Acori Tatarinowii extracts on gamma-aminobutyric acid type A receptor alpha 1 subunit brain expression during development in a recurrent seizure rat model

Extracts from Rhizoma Acori Tatarinowii （Grassleaf Sweetflag Rhizome, Shichangpu） have been shown to improve learning and memory, reduce anxiety, allay excitement, and suppress seizures. Rhizoma Acori Tatarinowii extracts interact with y-aminobutyric acid and activate the y-aminobutyric acid type A receptor, although few studies have addressed the precise effects of v-aminobutyric acid type A receptor al subunit. In the present study, y-aminobutyric acid type A receptor al subunit protein expression in the cerebral cortex and hippocampus, and pathological scores of brain injury, were significantly greater following recurrent seizures, but significantly decreased following treatment with Rhizoma Acori Tatarinowii extracts. These results indicated that Rhizoma Acori Tatarinowii extracts down-regulated y-aminobutyric acid type A receptor al subunit protein expression in the cerebral cortex and hippocampus and protected seizure-induced brain injury during development.

Study of Total Alkaloids from Rhizoma Coptis Chinensis on Experimental Gastric Ulcers

Objective： To study the effects of total alkaloids （TA） extracted from Rhizoma Coptis Chinensis on experimental gastric ulcer models. Methods： Four kinds of experimental ulcer models were established respectively by water-immersion stress, intragastric ethanol, acetic acid erosion, and pylorus ligation. The anti-ulcer effects of TA were evaluated, and compared with that of berberine （Bet） and cimetidine （Oim）. Results： TA showed significant inhibitory effects on ulcerative formation induced by water-immersion stress, intragastric ethanol, and pylorus l igation in dose-dependent manner, and showed therapeutic effect on acetic acid erosion-inducing ulcer, in comparison with the control group. The anti-ulcer activity of Bet was less than TA containing equal content of Bet. TA significantly reduced the free acidity, total acidity and total acid output, but didn＇t affect the gastric juice volume, gastric pepsin activity, adherent mucus quantity of stomach wall and free mucus dissolving in gastric juice. The suppressive activities of TA on gastric acid secretion didn＇t occur when it was administered into dodecadactylon at a dose of 360 mg/kg wt. Moreover, when compared with Oim, the inhibitory effect of TA on gastric acid secretion isn＇t proportional to the inhibitory effects on the formation of the 4 kinds of experimental ulcers. Conclusion： TA is a potent candidate in therapeutic drugs for treating gastric ulcer. Its anti-ulcer effective components and mechanism is not only related to Bet and inhibition of gastric acid, but also to other ingredients of TA and mechanism so far unknown.

Volatile components of Rhizoma Alpiniae Officinarum using three different extraction methods combined with gas chromatography-mass spectrometry

Volatile components from Rhizoma Alpiniae Officinarum were respectively extracted by three methods including hydrodistillation, headspace solid-phase microextraction （HS-SPME） and diethyl ether extraction. A total of 40 （hydrodistillation）, 32 （HS-SPME） and 37 （diethyl ether extraction） compounds were respectively identified by gas chromatography-mass spectrometry （GC/MS） and 22 compounds were overlapped, including β-farnesene, 7-muurolene, 2,6-dimethyl-6- （4-methyl-3-pentenyl）bicyclo[3.1.1]hept-2-ene, eucalyptol and cadina-1（10）, 4-diene and so forth, varying in relative contents. HS-SPME is fast, sample saving and solvent-free and it also can achieve similar profiles as those from hydrodistillation and solvent extraction. Therefore, it can be the priority for extracting volatile components from medicinal plants.

Effects of Cinnamon Granules on Pharmacokinetics of Berberine in Rhizoma Coptidis Granules in Healthy Male Volunteers

The effects of Cinnamon granules on pharmacokinetics of berberine in Rhizoma Coptidis granules in healthy male volunteers,and the compatibility mechanism of Jiao-Tai-Wan（JTW） composed of Rhizoma Coptidis granules and Cinnamon granules were investigated.The concentration of berberine in plasma of healthy male volunteers was determined directly by high performance liquid chromatogra-phy（HPLC） after an oral administration of Rhizoma Coptidis granules alone or combined with Cinnamon granules（JTW）.The plasma concentration-time curves of berberine were plotted.The data were analyzed with Drug and Statistics（DAS） 2.0 pharmacokinetic program（Chinese Pharmacology Society） to obtain the main pharmacokinetic parameters.The results showed that the plasma concentration-time curve of berberine was described by a two-compartment model.The Cmax,Tmax,t1/2 and CLz/F of berberine in Rhizoma Coptidis granules were 360.883 μg/L,2.0 h,3.882 h,119.320 L.h-1.kg-1 respectively,and those of berberine in JTW were 396.124 μg/L,1.5 h,4.727 h,57.709 L.h-1.kg-1 respectively.It was suggested that Rhizoma Coptidis granules combined with Cinnamon granules could increase the plasma concentration of berberine,promote berberine absorption and lengthen the detention time of berberine in healthy male volunteers.

Rapid analysis of components in Rhizoma Anemarrhenae by HPLC-DAD-MS and HPLC-DAD-TOFMS

A global quality control method based on high performance liquid chromatography(HPLC)coupled with diode array detection(DAD),single quadrupole mass spectrometry(MS)and time-of-flight mass spectrometry(TOFMS)was developed for simultaneous determination of seven major components(mangiferin,neomangiferin,timosaponin E1,timosaponin E,timosaponin BⅡ,timosaponin BⅢ,and timosaponin AⅢ)and identification of most components in extracts of Rhizoma Anemarrhenae(RA).HPLC analysis was performed on an Agilent SB-C18 column(4.6 mm×150 mm,5 μm)by gradient elution using acetonitrile and water-acetic acid(100∶0.05,v/v)as the mobile phase.Seven major components in RA were successfully separated.This quantitative method was fully validated in respect of the following performance criteria:linearity,precision,repeatability,stability,accuracy,limits of detection(LOD)and quantification(LOQ).A formula database of known compounds in RA was established,against which,most of the reported components in this herbal extract were identified effectively based on the extract masses acquired by TOFMS.This qualitative and quantitative method was successfully used to analyze the components in 10 batches of RA samples collected from different regions in China.This global quality control method,which consisted of HPLC-DAD-MS assay of seven major components and unambiguous identification of nineteen components,is suitable for routine quantification and comprehensive quality control of RA.

Qualitative and Quantitative Analysis for the Quality Control of Rhizoma Coptidis by HPLC-DAD and HPLC-ESI-MS

For quality control purpose, an approach of fingerprinting and simultaneous quantification of five major bioactive constituents of Rhizoma Coptidis was established via a high-performance liquid chromatograph coupled with a photodiode array UV detector（HPLC-DAD） and an electrospray ionization mass spectrometer（HPLC-ESI/MS） The compounds were identified on the basis of the comparison of their mass spectra with literature data and those of standard samples and quantified by the HPLC-DAD method. Baseline separation was achieved on an XTerra C18 column（5 μm, 250 mm×4.6 mm i. d.） with linear gradient elution of formate buffer（consisting of 0.5% formic acid, adjusted to pH=4.5 with ammonia） and acetonitrile（consisting of 0.2% formic acid and 0.2% triethylamine）. The me- thod was validated for linearity（r^2〉0.9995）, repeatability（RSD〈3.1%）, intra- and inter-day precision（RSD〈1.8%） with recovery（99.9%-105.1%）, limits of detection（0.15-0.35 μg/mL）, and limits of quantification（0.53-0.82 μg/mL）. The similarities of 32 batches of Rhizoma Coptidis and their classification according to their manufacturers were based on the retention time and peak areas of the characteristic compounds. The five compounds were selected for quality assessment ofRhizoma coptidis via partial least squares analysis（PLS）.

Comprehensive metabolic profiling of Alismatis Rhizoma triterpenes in rats based on characteristic ions and a triterpene database

Alismatis Rhizoma(AR)is widely used in Chinese medicine,and its major bioactive components,triterpenes,reportedly possess various pharmacological activities.Therefore,it is very important to study the metabolism of triterpenes in vivo.However,the metabolism of AR triterpene extract has not been comprehensively elucidated due to its complex chemical components and metabolic pathways.In this study,an ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry method,which was based on the characteristic ions from an established database of known triterpenes,was used to analyze the major metabolites in rats following the oral administration of Alismatis Rhizoma extracts(ARE).As a result,a total of 233 constituents,with 85 prototype compounds and 148 metabolites,were identified for the first time.Hydrogenation,oxidation,sulfate and glucuronidation conjugation were the major metabolic pathways for triterpenes in AR.In addition,the mutual in vivo transformation of known ARE triterpenes was discovered and confirmed for the first time.Those results provide comprehensive insights into the metabolism of AR in vivo,which will be useful for future studies on its pharmacodynamics and pharmacokinetics.Moreover,this established strategy may be useful in metabolic studies of similar compounds.

Review of Effects of Active Components of Gastrodiae Rhizoma on Central Nervous System

In order to explore the pharmacological effects of active components of Gastrodiae Rhizoma on the central nervous system,through consulting related literatures,this paper summarized the pharmacological effects of the active components of Gastrodiae Rhizoma.The pharmacological effects of the active components of Gastrodiae Rhizoma on the central nervous system include improving the ability of learning and memory,analgesia,sedation and hypnosis,anticonvulsant,anti-aging,anti-anxiety and neuroprotective effects.The active components of Gastrodiae Rhizoma have certain curative effect in the treatment of some diseases of the central nervous system.At present,it has been widely used clinically.There are no toxic and side effects at conventional doses,and it is worth popularizing and applying.

Active Ingredients and Mechanism of Action of Rhizoma Coptidis against Type 2 Diabetes Based on Network-Pharmacology and Bioinformatics

A pharmacological network of"component/target/pathway"for Rhizoma coptidis against type 2 diabetes(T2D)was established by network-pharmacology,and the active components of Rhizoma coptidis and its mechanism were explored.A literature-based and database study of the components of Rhizoma coptidis was carried out and screened by ADME paramcters.The targets of Rhizoma coptidis were predicted by the ligand similarity method.Related pathways were analyzed with databases,and software was used to construct a "component/target path" network.The mechanism was further confirmed by GEO database with R software.A total of 12 active components were screened from Rhizoma coptidis,involving 57 targets including MAPKI,STAT3,INSR,and 38 signaling pathways were associated with T2D.Related signaling pathways included essential pathways for T2D such as insulin resistance,and pathways that had indirect effect on T2D.It was suggested that Rhizoma coptidis may exert its effects against T2D through multi-component,multi-target,and multi-pathway forms.

Untiring Researches for Alternative Resources of Rhizoma Paridis

Rhizoma Paridis(RP,M®),a traditional Chinese medicine,is the rhizoma of Paris polyphylla var.yunnanensis(PPY)or P.polyphylla var.chinensis which are widely used as important raw materials for several Chinese patent drugs.However,the wild resources of these herbs have become less and less due to their slow-growing characteristics and previously excessive excavation.This review covers untiring investigations on alternative resources of RP by our research group over the past decades,including non-medicinal parts of PPY as well as other plants of Liliaceae and Liliflorae families.The arial parts of PPY and the whole plants of Trillium kamtschaticum might be alternative resources for RP based on the fact that they shared the same or similar saponins and bioactivities.