Simultaneous Determination of Six Active Components of Smilacis Glabrae Rhizome by UPLC-DAD

[Objectives] To determine the content of neoastilbin,astilbin,neoisoastilbin,isoastilbin,engeletin,and resveratrol in Smilacis glabrae rhizome. [Methods] The ultra-high-performance liquid chromatography method with diode array detection( UPLC-DAD) was developed. Analysis conditions as follows: waters Acquity UHPLC HT3 C18 chromatographic column( 2.1 × 100 mm),column temperature 30 ℃,mobile phase acetonitrile( A)-0.1% phosphoric acid( B) gradient elution,flow rate 0.2 mL/min,and detection wavelength 327 nm. [Results]Results indicated that the content of astilbin,neoastilbin,isoastilbin,neoisoastilbin,engeletin,and resveratrol in 10 samples of Smilacis glabrae rhizome was 1.313-6.1755 mg,0.013-3.699 mg,0.146-0.975 mg,0.137-0.7 mg,0.147-0.797 mg,0.077-0.112 mg,respectively. [Conclusions]This method is high efficient,convenient and accurate,and can be used as the method for quality control of Smilacis glabrae rhizome.

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.

Total glucosides of Rhizoma Smilacis Glabrae:a therapeutic approach for psoriasis by regulating Th17/Treg balance

Total glucosides of Rhizoma Smilacis Glabrae(RSG)are selective immunosuppressants that exhibit primary efficacy in the treatment of rheumatoid arthritis through targeted inhibition of activated T cells.In this study,we aimed to investigate the potential application of RSG in the treatment of psoriasis and elucidate its mechanism of action and material basis.Our findings revealed significant improvements upon administration of RSG in an imiquimod(IMQ)-induced psoriasis model.These improvements were characterized by a remarkable increase in the number of tail scales in mice and a substantial amelioration of skin erythema,ulceration,and flaking.By transcriptome sequencing and T-cell flow sorting assay,we identified notable effects of RSG on the modulation of various cellular processes.Specifically,RSG prominently down-regulated the Th17/Treg ratio in damaged skin tissues and reduced the proportion of G2 phase cells.Furthermore,RSG exhibited a stimulatory effect on the proliferation and differentiation of epithelial cells.Of particular interest,we discovered thatβ-sitosterol,sitostenone,stigmasterol,smiglanin,and cinchonain Ib displayed potent inhibitory effects on the IL-17-mediated inflammatory response in HaCaT cells.In summary,our study highlights the therapeutic potential of RSG in the treatment of psoriasis,attributed to its ability to regulate the Th17/Treg balance.These findings contribute to the development of new indications for RSG and provide a solid theoretical foundation for further exploration in this field.

正交试验优选土茯苓总黄酮的最佳提取工艺

[目的]优选土茯苓总黄酮的最佳提取工艺。[方法]在单因素试验的基础上,以土茯苓总黄酮的收率为指标,采用正交试验法优选土茯苓总黄酮的最佳提取工艺条件。[结果]土茯苓总黄酮的最佳提取工艺为:料液比为1∶20,乙醇浓度为60%,提取温度为95℃,回流提取2次,每次2 h;在此条件下,总黄酮的收率为8.34%。[结论]该工艺操作简单、稳定、可行。

小花棘豆中毒对家兔脏器指标的影响

为探讨小花棘豆(Oxytropis glabra DC)对家兔脏器指标的影响,试验挑选健康22只,分为两组,其中,试验组12只[按照10 g/(d·kg体重)的剂量饲喂小花棘豆干粉];对照组10只(饲喂青干草和少量精料),试验期为70 d,试验结束后取其脏器,计算脏器指数。结果显示,与正常对照相比,试验组家兔脑指数、肝脏指数、肾脏指数、肾上腺指数、睾丸指数、卵巢指数和肺脏指数增大,心脏指数、脾脏指数和胰脏指数下降,部分指标差异达到显著或极显著(P<0.05或P<0.01),与小花棘豆中毒相关性最强的指标为脑指数;但小花棘豆中毒对家兔胃肠道指标无明显影响。结果表明,小花棘豆中毒导致家兔肝、肾、脑等实质性器官损伤。

药用植物草珊瑚AFLP反应体系的建立

从草珊瑚(Sarcandra glabra)的幼嫩叶片中提取基因组DNA,建立草珊瑚AFLP反应体系,对AFLP反应体系中模板DNA的浓度、基因组DNA的双酶切时间、预扩增产物的稀释倍数和引物组合的筛选等关键因素进行摸索。优化的草珊瑚AFLP反应体系为模板DNA的用量20 ng/μL、酶切反应时间4 h、预扩增产物稀释15倍,初步筛选出8对较为适合草珊瑚AFLP分析的引物组合。

三角梅在咸宁市不同环境条件下越冬的抗寒表现

三角梅(Bougainvillea glabra Choisy)在咸宁市因冬季低温进入被迫休眠,同品种同类型的三角梅在不同环境所致的不同开花状态下进入休眠的早晚不同,开花状态的三角梅进入休眠相对较晚。三角梅在越冬期蒸腾强度呈现逐渐减弱的趋势,越冬期因叶片蒸腾和土面水分蒸发使盆土相对含水量日下降0.5%左右。当气温低于-4℃时三角梅受冻;如果没有叶片的防冻保护,三角梅在-2℃就受到低温和寒风的危害,受害程度与风力的大小有关。

小花棘豆中毒对和田羊内脏器官α-甘露糖苷酶的影响

探讨小花棘豆(Oxytropis glabra DC)中毒对和田羊内脏器官α-甘露糖苷酶(AMA)的影响,进一步揭示小花棘豆的毒性作用机理。将12只和田羊随机分为3组,即对照组、试验Ⅰ组和试验Ⅱ组。试验组分别按10 g/kg和20 g/kg的剂量饲喂小花棘豆,饲喂至出现典型中毒症状。屠宰后每组随机采集试验羊的肝脏、肾脏、脾脏、心脏和肺脏,检测和田羊内脏器官AMA活性及表达的变化。结果表明,和田羊肝脏、肾脏、脾脏、心脏和肺脏中高尔基体α-甘露糖苷酶Ⅱ(AMA1)和溶酶体α-甘露糖苷酶(AMA2)均有表达。试验Ⅰ组和田羊各器官的AMA2的表达均与对照组差异不显著(P>0.05),试验Ⅱ组和田羊肝脏和肾脏AMA2的表达显著低于对照(P<0.05),但其余器官AMA2的表达均与对照组差异不显著(P﹥0.05);试验Ⅱ组和田羊肝脏和肾脏AMA1的表达极显著低于对照(P<0.01),脾脏AMA1的表达显著低于对照(P<0.05);试验Ⅰ组和田羊肝脏、肾脏AMA1的表达显著低于对照(P<0.05),其余器官AMA1的表达均与对照组差异不显著(P>0.05)。不同剂量小花棘豆均可降低和田羊内脏中AMA活性及其m RNA表达量,肝脏和肾脏是其主要作用的靶区。

小花棘豆中毒对绵羊运氧能力和抗氧化损伤的研究

探讨小花棘豆(Oxytropis glabra DC)对绵羊抗氧化能力的影响,揭示小花棘豆对绵羊的毒性作用机理。将16只绵羊随机分为4组,即对照组、试验Ⅰ组、试验Ⅱ组和试验Ⅲ组,3个试验组分别按照每千克体重5、10、20 g的剂量饲喂小花棘豆,至典型临床症状出现,通过采血检测绵羊红细胞和抗氧化指标的变化。结果显示,与正常对照相比,中毒绵羊血液红细胞数(RBC)、红细胞压积(HCT)、血红蛋白(HGB)、平均红细胞血红蛋白含量(MCH)、平均红细胞体积(MCV)、甘油三酯(TG)、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、黄嘌呤氧化酶(XOD)活性显著下降,红细胞分布宽度(RDW)、总胆固醇(TC)、丙二醛(MDA)、一氧化氮(NO)、一氧化氮合酶(NOS)、低密度脂蛋白(LDL-C)和极低密度脂蛋白(VLDL-C)含量均显著上升。结果表明,小花棘豆中毒可显著影响绵羊血液运氧和抗氧化能力,并影响绵羊的脂代谢。

三角梅在咸宁地区盆栽育花技术初探

三角梅(Bougainvillea glabra Choisy)在咸宁地区盆栽需采用人为催花措施方能繁花似锦。将三角梅在春季用小盆盆栽,夏季进行干旱胁迫以抑制营养生长,达到一定时间或程度时随即扩盆松土、施肥浇水,可取得较好的催花效果。此外,摘心、拉枝、强光高温对三角梅具有一定的催花效果。培育三角梅整齐的花枝花冠还需适时轻剪和秋季保温。

Approaches and Perspectives of the Westward Spread of Traditional Chinese Medicine: A Case Study of the Radicis Chynae

Some Western scholars have re-examined the concept of“Chinese medicine”and its knowledge system under the influence of global history research methods in recent years,in an attempt to understand the factors that led to the spread of Chinese medicine around the world,and what kind of Chinese medicine is constituted outside of China.Thus,researchers have studied the initial stage of traditional Chinese medicine(TCM)’s entry into the western world,tracing its roots and observing new knowledge systems formed in the process of cross-cultural communication.Responding to the research of Western scholars,this paper studies the Radicis Chynae(《中国根书简》Letters on Chinese Root),a monograph written by Andreas Vesalius,a famous European anatomist who lived in the 16th century.The author of this article examined the understanding and interpretation of Chinese medicine by the European intelligentsia from 16th to 19th century,investigated the influence of the westward spread of TCM on the scientific revolution and medical progress in Europe,and analyzed its relationship with the rise of Sinology in Europe.This article discusses the knowledge interaction between Chinese medicine and the formation of modern European medicine from the perspective of global history and cross-culture.

A Novel Phenylpropanoid-substituted Catechin Glycoside and a New Dihydrochalcone from Sarcandra glabra

A novel phenylpropanoid-substituted catechin glycoside glabraoside A 1 and a new dihydrochalcone 3＇-（7＂-allylphenyl）-2＇,4＇,4＂-trihydroxy-6＇-methoxydihydrochalcone 2 were isolated from the herbs of Sarcandra glabra. Their structures were elucidated on the basis of spectroscopic analyses and chiroptical methods.

A Flavonol Glycoside from Smilax glabra

A new flavonol glycoside, named neosmitilbin was isolated form the rhizome of Smilax glabra. Its structure and absolute configuration were elucidated on the basis of spectroscopic studies.

Role and Mechanism of Action of Exogenous Salicylic Acid and Sodium Molybdate in Improving Cold Tolerance of Bougainvillea glabra

This study was conducted to comprehensively evaluate the effects of salicylic acid and sodium molybdate on cold tolerance of an ornamental plant Bougainvillea glabra and to provide a theoretical guidance for landscape maintenance.B.glabra plants were treated with 0.5 mmol/L salicylic acid and 2.0 μmol/L alone or in combination,and then exposed to low temperature stress before physiological indices were measured.The results showed that all salicylic acid and sodium molybdate treatments reduced the relative conductivity and malondialdehyde( MDA) content of B.glabra to varying extents under the stress of low temperature,and more significant effect was achieved by using the two agents in combination.Oxygen free radicals production rate increased with decreasing temperature from 20 to 6 ℃,but declined with temperature decreasing from 3 to-3 ℃.The SOD activity of the control( CK) was significantly lower than that of other treatments at 0 and-3 ℃.The treatments with salicylic acid and sodium molybdate alone and in combination increased POD activity of B.glabra plants,especially at 0 ℃,as the POD activity of treatments T1,T2 and T3 was significantly higher than that of CK at 0 ℃.In addition,under low temperature stress,the contents of soluble sugar,starch and proline increased initially and decreased subsequently with temperature decreasing.The soluble sugar content at 3 ℃,starch and proline contents at 0 and-3 ℃ in treatments with salicylic acid and sodium molybdate alone and in combination were significantly higher than those of CK.All above results proved that salicylic acid and sodium molybdate are able to improve cold tolerance of B.glabra,and better effect can be achieved by using them together.

Study on chromatographic fingerprint of sarcandra glabra (Thunb.) by microwave-assisted extraction coupled to HPLC/DAD

Microwave-assisted extraction(MAE)was used for extraction of effective components of sarcandra glabra(Thunb.),and then chromatographic fingerprint of sarcandra glabra(Thunb.)was studied by high performance liquid chromatography/diode array detector(HPLC/DAD).The conditions of MAE were optimized by an orthogonal experiment,and then the authentication and validation of the chromatographic fingerprint were conducted.Nine peaks were identified as common peaks in the fingerprint chromatograms,and isofraxidin was considered as a reference compound and quantified.Relative standard deviations of retention time and peak area of each component were less than 3% and 8%,respectively.Similarity and difference analysis were conducted by use of PCA and relation coefficient.Twenty batches of sarcandra glabra(Thunb.)samples from two different producing areas could be classified into two different groups in the PCA model.The results showed that MAE-HPLC/DAD method was simple,efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra(Thunb.),which could provide more reliable and precise information for quality evaluation.

Hydroalcoholic extract of licorice(Glycyrrhiza glabra L.) root attenuates ethanol and cerulein induced pancreatitis in rats

Objective:To evaluate the therapeutic potential of hydroalcoholic extract of licorice root against ethanol and cerulein induced chronic pancreatitis in rats.Methods:The phytochemical profile of hydroalcoholic extract of licorice root was determined by high performance liquid chromatography(HPLC)and gas chromatography coupled to mass spectrometry(GC-MS).Chronic pancreatitis was induced in male albino Wistar rats by feeding them a diet containing ethanol(0%-36%of total calories)for 4 weeks and cerulein(20μg/kg b.wt,i.p.)thrice a week for 3 weeks.Lipase and amylase in serum,lipid peroxides and antioxidants including reduced glutathione,glutathione peroxidase,superoxide dismutase and catalase in pancreas were determined.Inflammatory response was measured by myeloperoxidase in the pancreas,caspase-1 and the concentrations of IL-1 as and liver was carried oαand IL-18 in serum.Moreover,histological evaluation of the pancreut.Results:Different flavonoids and saponins were identified in the hydroalcoholic extract of licorice root through HPLC and GC-MS.A marked increase in the levels of serum lipase,amylase,lipid peroxides,caspase-1,myeloperoxidase,IL-1 in the levels of antioxidants were observed after ethanoα,and IL-18 and a marked decrease l and cerulein administration.Treatment with hydroalcoholic extract of licorice root attenuated these changes.In addition,histological observation confirmed the protective effect of the extract in the pancreas and liver against inflammatory changes induced by ethanol and cerulein.Conclusions:The licorice root extract attenuates ethanol and cerulein induced pancreatitis in rats probably due to its antioxidant phytonutrients since ethanol and cerulein-induced production of reactive oxygen species contributes to severe inflammation in the pancreas.

Ethyl acetate extract of Smilax glabra Roxb roots and its major active compound astilbin promote osteoblastogenesis in vitro by upregulating bone cell differentiation-associated genes

Objective:To investigate the osteoblastogenic activity of the ethyl acetate(EtOAc)extract of Smilax glabra Roxb roots and its major active compound astilbin.Methods:Astilbin was isolated from EtOAc extract using silica gel chromatography combined with fraction crystallization.Chemical structure of astilbin was determined by analysis of the spectroscopic data in comparison with the literature.MTT method was used to detect the toxicity.Alkaline phosphatase(ALP)activity was determined by the spectrophotometric method at 405 nm using p-nitrophenyl phosphate as a substrate.Calcium deposition was stained with alizarin red-S,distained with cetylpyridium chloride,and quantified at 562 nm.In silico model for astilbin-ALP interaction was analyzed using AutoDock 4.2.6.The changes in expression of osteoblast differentiation related genes were determined using quantitative real-time PCR.Results:Both the EtOAc extract and astilbin had no toxicity toward osteoblast MC3T3-E1 cells at 5.0,10,25,and 50μg/mL.At 25μg/mL,they enhanced ALP activity and mineralization of osteoblasts up to 30%and 55%for the EtOAc extract and 22%and 41%for astilbin,respectively.Molecular docking analysis of astilbin-ALP interaction revealed Arg167,Asp320,His324,and His437 were key residues participating in hydrophobic interaction;meanwhile,His434 and Thr436 residues were involved in hydrogen bond formation in the active site of human tissue-nonspecific ALP.Moreover,the expression level of genes opn,col1,osx,and runx2 were up-regulated in astilbin treated samples with the fold changes as 2.2;3.7;4.1;2.3,respectively at 10μg/mL(P<0.05).Conclusions:The EtOAc extract and its major compound astilbin exhibit osteoblastogenic activity by up-regulating important markers for bone cell differentiation.It could be a new and promising osteogenic agent with dual actions for therapeutic applications.

2D NMR Spectroscopic Studies of the Toxic Alkaloid from Oxytropis Glabra D C.

The Oxytropis, legume genus, is native in the northwest of China, and some of its members have been reported to be toxic to grazing livestock. In the course of studying toxic reason of this legume, a toxic alkaloid was isolated from Oxytropis glabra D C. grown in the Inner Mongolian Autonomous Region. The LD+（50） of （—）-thermopsine（Ⅰ） was shown to be 89.98 mg/kg（in mice）. This paper reports an approach to the structure elucidation of（-）-thermopsine by combining 2D NMR[;H-;H and;H-;C （one-bond and long-range） COSY] and other physicochemical methods. The stereochemistry of compound Ⅰ is also discussed.

Development of a validated UPLC-qTOF-MS/MS method for determination of bioactive constituent from Glycyrrhiza glabra

An ultra-performance liquid chromatography quadrupole time of flight mass spectrometry （UPLC-qTOF-MS/MS） method was developed and validated for the simultaneous determination of glycyrrhizin and glycyrrhetic acid. These analytes were separated on a reverse phase C18 column using a mobile phase of acetonitrile：2% acetic acid in water （75：25, v/v） with a flow rate of 200 μL/min. The qTOF-MS was operated under multiple reaction monitoring （MRM） mode using the electrospray ionization （ESI） technique with positive ion polarity. A comparison of three different extraction techniques i.e. accelerated solvent extraction （ASE）, extraction under ultrasonic waves （USW） and the classical extraction by percolation （CE） method was done and quantification of these extracts was also carried out by the proposed method.

Preparation of Polyclonal Antibody Against Swainsonine and Its Effects in Treatment of Oxytropis glabrapoisoned Rabbits

This study was conducted to optimize immunization program to gain the serum against swainsonine （SW） with high titer and to analyze the effects of polychinal antibody in treatment of loco disease of rabbits. Eighteen rabbits were randomly separated into control group, poisoning group and curing group, with six rabbits in each group. Rabbits in poisoning group and curing group were fed 10 g/（kg ~ d） powder of Oxytropis glabra DC every morning for 70 d till the rabbits in poisoning group began to die. From the 21＇t to 24＇h d of O. glabra administration, each of the rabbits in curing group was injected with 1 mL of polyclonal anti- body against SW every day. Blood samples were collected before O. glabra challenge, and then once every 7 d for the serological and immunological evaluation of serum. The resuhs showed that the serum alkaline phosphatase （AKP） activity and SW content in poisoning groups were significantly higher than those in control group on the 7th d （P 〈0.05 ） ; the serum aspartate transaminase （AST） and lactate dehydrogenase （LDH） activity in poisoning groups were significantly higher than those in control group on the 14th d （P 〈 0.05 ）, while the serum ct - mannosidase （AMA） and rate of E-rosette formation in poisoning groups were signifi- cantly lower than those in control group （ P 〈 0.05 ） ; the serum blood urea nitrogen （BUN）, creatinine （ CRE ） and glucose （ GLU ） concentration in poisoning groups were significantly higher than those in control group on the 21＇t d （P 〈 0.05 ）. By injecting serum against swainsonine, the serum AKP, LDH, AST, alanine transaminase （ALT） activity and contents of BUN, CRE, GLU, SW were reduced and serum AMA and rate of E-rosette formation were increased in curing group, by comparing with those in poisoning group. These results in this experiment indicated that polyclonal antibody against SW had good value of preventing from O. glabra DC.