Effect of pH on Antioxidant and Phytochemical Activities of Mulhatti Roots (Glycyrrhiza glabra L.)

Glycyrrhiza glabra L.is the most widely used herb in the ancient history of Ayurvedic medicine,as a medicinal value as well as an aromatic herb,and it is commonly known as Mulhatti.Mulhatti roots are useful for medically and are also a good source of phytoproducts and secondary metabolites present in Mulhatti roots are triterpenoid saponin,glycosides,glycyrrhizin,prenylated biaurone,licoaagrone,7-acetoxy-2-methylisoflavone,4-methylcoumarin,liqcoumarin,glycyrrhetinic acid,quercetin,liquiritigenin,isoliquiritigenin,etc.This study was carried out to study the evaluation of phenolic compounds,2,2-diphenyl-1-picrylhydrazyl(DPPH)free radical activity and general antioxidant capacity of water extracts of Mulhatti roots prepared at different pH values,namely 2,4,7 and 9.Data have shown great differences in terms of results.Most of the phenolic compounds are at pH 7(19.25),followed by pH 9(17.25),pH 2(14.62)and pH 4(8.89 mg GAE/g),respectively.Similarly,the flavonoid data also showed variations,the maximum has been present in pH 2(5.39),then pH 7(3.02),pH 9(1.79)and pH 4(1.40 mg CE/g),respectively.The value for DPPH IC50 free radical scavenging activity was the lowest at pH 7(82.22),followed by pH 2(110.40),pH 4(111.99)and pH 9(146.24μg/mL)and IC50 reference standard(ascorbic acid)was 59.52μg/mL in distilled water.The total capacity of the antioxidant was the highest at pH 2(9.93)followed by pH 4(5.54),pH 7(5.34)and pH 9(4.23 mg AAE/g).According to current research,pH 7 is the best for phytochemicals as well as antioxidants that catch harmful radicals.

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.

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.

Molecular docking analysis to validate the efficacy of phytocompounds of traditional Siddha herb Glycyrrhiza glabra against enzyme tyrosinase for the treatment of vitiligo

Background: Siddha system is one of the ancient medical systems which originated in Indiamore than 4000 years ago had given many formularies and treatment methods for variousdiseases especially skin diseases and autoimmune diseases. One of the most well-knownmembers of the Fabaceae family and a popular medicinal plant is Athimathuram(Glycyrrhiza glabra). Glycyrrhiza glabra root powder (Athimathuram chooranam) fromSiddha herbal formulation mentioned in the texts to treat vitiligo. The study is aimed toexecute the In Silico computational studies of phytoconstituents of Siddha herbAthimathuram chooranam (Glycyrrhiza glabra) against Tyrosinase enzyme for the treatmentof vitiligo. Method: Making use of Auto Dock 4, docking calculations were performed.Gasteiger partial charges were applied to the ligand atoms. In order to simulate docking, theSolis & Wets local search approach and the Lamarckian genetic algorithm were employed.The ligand molecules’ initial locations, orientations, and torsion angles were determined atrandom. During docking, all rotatable torsions were freed. Results: By interacting withamino acids located on the active site of the tyrosinase enzyme, bioactive substances likeGlycyrrhizin, Liquirtin, Glabridin, Umbelliferone, and Glabrene included in the Siddhaformulation exhibit substantial binding activity against the target protein. Conclusion: Itwas concluded that these compounds might exert promising anti-vitiligo properties.

SCoT与EST-SSR标记检测甘草属(Glycyrrhiza L.)植物遗传多样性的比较研究

为探讨SCoT、EST-SSR标记技术在甘草属植物遗传多样性研究的适用性,采用SCoT、EST-SSR分子标记对甘草属8自然居群的4种1变种共计14个个体进行基因组DNA的多态性检测。实验筛选出的18对SCoT通用引物共扩增出313条带,其中312条为多态性条带,平均多态率(PPB)为99.68%,平均有效等位基因数(Ne)和位点品均期望杂合度(He)等遗传参数与EST-SSR标记无显著差异。多态性检测效率高的标记为SCoT,E=17.333,Ai=21.145,高于EST-SSR,E=6.909,Ai=8.719。SCoT与EST-SSR标记聚类分析结果趋势相似,均将14份材料划分为3组,与基于形态学分类结果一致。2种标记相比,EST-SSR(0.820)产生的平均相似系数范围大于SCoT(0.785),EST-SSR标记个体间遗传差异检测能力略高于SCoT。结果表明SCoT和EST-SSR两种标记均可揭示甘草属种间与种内的遗传多样性水平以及亲缘关系,是进行甘草属植物自然群体的遗传结构、种间基因渐渗等研究的有效分子标记。其中,SCoT标记多态性高,信息量大,更适于甘草属植物种质资源的遗传多样性分析;个体间遗传差异检测上EST-SSR标记效果更佳,更利于疑难种鉴定及亲本分析。

Use of Support Vector Regression Based on Mean Impact Value Model to Identify Active Compounds in a Combination of Curcuma longa L.and Glycyrrhiza extracts

A support vector regression based on the mean impact value (MIV) model was constructed to identify the bioactive compounds inhibiting proliferation of HeLa cells in a combination of turmeric (Curcuma longa L.) and liquorice (Glycyrrhiza) extracts. The quantitative chemical fingerprint from 50 batches of turmeric and liquorice extracts was established using high performance liquid chromatography hyphenated to an ultraviolet visible detector. Qualitative results were obtained using ultra performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry. A total of 46 peaks (peaks 1–15 from turmeric and 16–46 from liquorice) were selected as “common peaks” for analysis. The inhibitory effect of the combined extracts on HeLa cells was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. It was found that 15 compounds (peaks: 8, 12, 30, 24, 46, 11, 14, 9, 3, 1, 44, 18, 7, 45 and 43) possessing high absolute MIV exhibited a significant correlation with the cytotoxicity against HeLa cells; most of these have already been confirmed with potential cytotoxicity in previous research. The important potential application of the present model can be extended to help discover active compounds from complex herbal medicine prior to traditional bioassay-guided separation. It is considered that this could be a useful tool for re-developing herbal medicine based on the use of these active compounds. © 2017, Tianjin University and Springer-Verlag Berlin Heidelberg.

INHIBITORY EFFECT OF GLYCYRRHIZA URALENSIS FISH AND CHELIDONIUM MAJUS L ON GASTROCARCINOGENESIS . INDUCED BY N-METHYL-N' -NITRO-N-NITROSOGUANIDINE

In order to investigate the antagonistic effect of Glycyrrhlza Uralensis Fish (GUF) and Chelidonium maJus L (CML) on gastrccarcinogenesls Induced by MNNG in Wastar rats, we treated the rats with MNNG alone (group 1) and with MNNG plus GUF and CML (group 2 and 3) respectively. The Incidence of Infiltrating adenocarcinoma of the glandular stomach and duodenum in group 2 was significantly lower than that in group 1 (26.7% vs. 67.8%). The differentiation and aggresslvenees of carcinomas occured in group 2 were much better and mild than those in group 1. Present study also demonstrated that the Inhibitory effect of CML on proliferation of human stomach carcinoma cell line MGC-803 was very remarkable; in addition, GUF and CML were able to antagonise the mutagenlc activation of MNNG. These results suggest that GUF and CML may be empoyed In prevention of gastric carcinoma.

NaCl与Na_2CO_3胁迫对光果甘草幼苗叶片叶绿体色素含量的影响

以水培光果甘草幼苗作为试验材料,分别进行不同浓度NaCl[0.2%,0.5%,0.8%(w/v)]与Na2CO3[0.01%,0.05%(w/v)]胁迫处理,以完全营养液培养作为对照,在处理第5天后测定各处理叶片中叶绿素a,叶绿素b,类胡萝卜素含量并计算叶绿素a与叶绿素b的比值。结果表明:在NaCl胁迫处理下,甘草叶片chla、chab、car含量均以0.2%NaCl为最高,之后随盐浓度升高而下降,而cha/chlb值则随盐浓度的升高而升高,在Na2CO3处理下,甘草叶片chla、chlb、car含量随Na2CO3胁迫处理浓度的升高而升高,而cha/chlb值则呈现下降的变化趋势。无论是NaCl处理还是Na2CO3处理各处理浓度间上述指标差异均达显著或极显著水平。

黄药子配伍甘草对LO2细胞膜通透性影响的研究

目的:研究甘草配伍能减轻黄药子对人肝细胞(LO2)膜通透性的影响。方法:激光扫描共聚焦显微镜(LSCM)对经二乙酸荧光素(FDA)染色后的LO2细胞进行实时动态荧光强度检测。通过比较荧光强度的降低率来考察黄药子及其甘草配伍后含药血清在短时间内对细胞膜通透性的影响。结果:LSCM记录细胞荧光强度变化,在10 min扫描停止时,黄药子各组荧光强度降低率与空白组有统计学差异(P<0.05),而甘草配伍组对荧光强度的影响较黄药子组有统计学差异(P<0.05),与空白组无差异(P>0.05)。结论:黄药子含药血清短时间内即影响肝细胞膜通透性,甘草配伍后可明显减弱损伤作用,黄药子引起肝损伤的机制可能是其对肝细胞膜的直接损伤作用,甘草配伍减毒作用与此途径有关。

银屑冲剂中原儿茶酸的HPLC法测定

目的:建立银屑冲剂中原儿茶酸的含量测定方法。方法:采用反相高效液相色谱法,色谱柱:C_(18)(250 mm×4.6 mm,5 μm);流动相:甲醇-水-冰醋酸(15:85:0.5);流速:1.0 mL.min^(-1);检测波长:260 nm。结果:原儿茶酸在0.1032～0.516μg范围内呈良好线性关系,r=0.99996,平均回收率97.27%,RSD为1.4%。结论:建立的方法简便可行,重现性好,提高了银屑冲剂质量控制方法。

马钱子与甘草配伍前后士的宁和马钱子碱的HPLC分析

目的:研究马钱子配伍甘草对马钱子中主要生物碱的影响。方法:采用高效液相色谱法分别对马钱子配伍甘草前后主要生物碱成分士的宁和马钱子碱的含量进行测定。结果:马钱子配伍甘草后其主要生物碱士的宁和马钱子碱的含量均有不同程度的降低。结论:马钱子配伍甘草可降低毒性,为进一步阐明马钱子合理配伍用药提供了科学实验依据。

新疆光果甘草黄酮对人肝癌Bel-7402细胞增殖的抑制活性研究

目的探讨新疆光果甘草(Glycyrrhiza glabra L)总黄酮及单体成分光甘草定(glabridin,Gb)对人肝癌Bel-7402细胞增殖的抑制活性。方法采用乙醇提取法从光果甘草制备总黄酮提取物(GF-A);经聚酰胺富集法精制而制备黄酮含量高于50%的标准化提取物(GF-B);从GF-B中纯化制备单体成分光甘草定(Gb);以人肝癌Bel-7402细胞株作为体外模型,用MTT法测定各样品对肝癌细胞增殖的抑制活性。结果制备和鉴定3种提取物:GF-A(总黄酮含量31.18%)、GF-B(含量52.5%)和Glabridin单体;3种物质对人肝癌Bel-7402细胞的增殖显示明显的抑制活性,其浓度在25～250μg/mL范围内对肿瘤细胞的最高抑制率分别达67.92%、84.28%和90.11%。结论人肝癌Bel-7402细胞对光果甘草总黄酮的抗癌活性具有一定的敏感性;单体Glabridin可能是光果甘草总黄酮中抗癌活性较高的有效成分之一。

UPLC-Q-Exactive Orbitrap-MS法分析光果甘草地上部分黄酮类成分

[目的]对光果甘草地上部分(茎、叶)的黄酮类化学成分进行分析,为进一步开发利用光果甘草地上部分及阐明其药效物质基础提供科学依据。[方法]将光果甘草地上部分醇提样品水沉,分为水溶性、脂溶性部位;采用超高效液相色谱-四级杆-静电场轨道阱联用(UPLC-Q-Exactive Orbitrap-MS)法检测样品,使用Xcalibur软件处理分析数据,将得到的成分分子质量、多级图谱碎片信息结合文献、数据库及标准品进行指认。[结果]从光果甘草地上部分共检测出100个黄酮类成分,有97种在光果甘草地上部分被首次发现;其中水溶性样品检测出61种,脂溶性样品检测出69种。[结论]通过UPLC-Q-Exactive Orbitrap-MS法,对光果甘草地上部分建立了快速、准确的定性方法,同时证实黄酮类成分含量丰富,为进一步开发利用提供依据。

HPLC测定光果甘草中光甘草定的含量

目的:采用HPLC测定光果甘草粗黄酮浓缩物中光甘草定的含量。方法:以FL2200-II为高效液相色谱仪,采用Ultimate XB-C18色谱柱,以水和乙腈为流动相,进行梯度洗脱,检测波长280nm,流速1.0mL.min-1。结果:光甘草定在9.5～475μg.mL-1内线性良好,r=0.9998;测得某光果甘草粗黄酮浓缩物中光甘草定的平均含量为3.58%,RSD=1.23%,平均加标回收率99.04%,RSD=0.26%。结论:利用反相高效液相色谱对甘草粗黄酮浓缩物中的光甘草定进行含量测定,方法简便,准确度高,重现性好,可应用于光甘草定的含量测定。

甘草提取物对雷公藤甲素损伤后人肝L-02细胞中UGT1A、MRP2表达的影响

目的:考察甘草提取物(GE)对雷公藤甲素(TP)损伤后人肝L-02细胞中尿苷二磷酸葡萄糖醛酸转移酶1A(UGT1A)、多药耐药相关蛋白2(MRP2)表达的影响,研究甘草对TP的减毒机制。方法:采用MTT法测定0(空白对照)、40、80、160 nmol/L TP分别培养L-02细胞12、18、24 h后的细胞存活率。将L-02细胞分为空白对照组(空白培养基)、模型对照组(80 nmol/L TP)和GE预处理组(分别以30、60、90 mg/L GE预处理24 h后再加入80 nmol/L TP),继续培养18 h后,采用MTT法检测各组的细胞存活率。在上述分组(GE预处理组的GE浓度为60 mg/L)基础上增设利福平(RIF)组(阳性对照,10μmol/L RIF预处理24 h后再加入80nmol/L TP),培养24 h后,检测各组细胞中UGT1A、MRP2蛋白表达。结果:TP对细胞活性的抑制作用与浓度和时间呈正相关。与空白对照组比较,模型对照组的细胞存活率明显降低(P<0.05),细胞中MRP2蛋白表达明显减弱(P<0.01)。与模型对照组比较,30、60、90 mg/L GE预处理组的细胞存活率均明显升高(P<0.01);60 mg/L GE预处理组细胞中UGT1A和MRP2蛋白表达均明显增强(P<0.01)。结论:GE预处理可减轻TP对人肝L-02细胞的损伤,其减毒机制可能与提高细胞中UGT1A、MRP2的表达有关。

Effect of the Nutritional Supplement VIUSlD Vet on the Productivity of Fattening Pigs in a Low-Input Farming System

The objective of the study was to evaluate the effect of the nutritional supplement VIUSID vet on the mortality and the productivity behaviour of fattening pigs in a low-input farming system. Two experiments were carried out at a pig farm in the municipality of Jatibonico, Sancti Spiritus province, Cuba. The farm has a low level of specialization, pigs are fed using local resources such as sugarcane （Saccharum officinarum） juice and locally produced concentrated feed. In both studies, the treatment consisted in administering together with the feed 2.0 g of VIUSID vet per Kg of feed. Results showed that in low-input rearing conditions, supplying VIUSID vet significantly reduced deaths （17.65%） in the pigs treated, favoured leukocyte and lymphocyte counts, and significantly improved weight gain （16.33%） and feed efficiency （14.41%）.

Effect of Hoechunyangkyeok-San Extract on Melanogenesis

Forsythia fructus has been shown to have antioxidative, anti-inflammatory, antibacterial, anti-aging and whitening effects. Hoechunyangkyeok-san (Forsythia viridissima-prescription) is a traditional herbal medicine, which has been clinically used for treating febrile and inflammatory disorders. This work was carried out to investigate the skin whitening effects of Forsythia viridissima-prescription extract (a hydrolyzed extract of Hoechunyangkyeok-san: SID White HYC) on skin. The effects of SID White HYC were assessed the melanin contents in B161 melanoma cells and the pigmented equivalent with HMB45 and Fontana Masson staining in 3D skin model. Then, we examined the expression of major pigment enzymes regulating melanin synthesis and melanosome transport related proteins in B16F1 cells. SID White HYC significantly inhibited the melanin synthesis (56.7% and 30.6% inhibition at 100 μg/mL, intracellular and secreted, respectively) in B16F1 cells and 3D skin model. In addition, western blotting analysis showed that SID White HYC reduced the expression of melanin synthesis and melanosome transport related proteins in B16F1 cells. In clinical trials, the cream containing 0.05% SID White HYC showed skin depigmentation effect without any irritation. These results suggest that SID White HYC may be useful inhibition of melanogenesis and melanosome transport. Therefore, SID White HYC may have potential as a skin-whitening ingredient in cosmetics.

多基原甘草质量标准探讨

目的分析多基原甘草质量现状及存在的问题,探索制约因素并提出解决方案。方法通过和企业座谈、实地调研、文献查询及甘草质量标准研究工作,分析当前甘草产业现状、药用标准问题及发展契机。结果以《中华人民共和国药典》(2020年版)含量测定标准,我国胀果甘草、光果甘草中甘草苷含量不合格率偏高,但3种基原的甘草存在其他具有药理作用的黄酮类化学成分,仅以甘草苷作为质量控制指标评价不够全面。结论建议进一步提高和完善《中华人民共和国药典》中胀果甘草和光果甘草质量标准,采用多种黄酮类成分作为甘草质量评价指标,进而推进我国甘草种植业发展。

中药甘草渣制备羧甲基纤维素的工艺优化及其应用

本研究旨在利用光果甘草渣合成羧甲基纤维素(CMC),以减少甘草资源的浪费并提升其应用价值。以低共熔溶剂提取后的新疆光果甘草渣为原料,采用两步法合成了CMC,考察不同合成条件(NaOH质量分数、氯乙酸钠与甘草渣质量比、醚化温度和醚化时间)对合成CMC取代度的影响并优化了反应条件,对其理化性质进行结构表征,并利用沙土保水实验和草莓保鲜实验验证甘草渣CMC的应用价值。结果表明,最优条件为NaOH质量分数50%、氯乙酸钠和甘草渣质量比1.2∶1、醚化温度60℃、醚化时间3 h,该条件下制得的CMC取代度达到0.76。在沙土中添加制备的CMC可以显著提高沙土的持水能力,减少水分蒸发,48 h后保水率可达45.7%。所制备的CMC生物降解性良好,同时还可以促进微生物的生长。采用制备的CMC膜溶液包裹草莓可将草莓的保质期延长至7 d,具备良好的应用前景。

光果甘草生殖生物学特性的初步研究

对光果甘草的花部特征、花粉活力、柱头可授性和花粉-胚珠比(P/O)进行了初步研究.结果表明:(1)光果甘草雄蕊异型,雌雄蕊异位,有利于避免自花授粉;(2)开花当天花粉活力在上午12:00最高,为91.8%,14:00迅速下降,16:00到达最低,16:00以后逐渐上升;(3)柱头可授性可持续4 d,以开花当天可授性最强;(4)花粉-胚珠比(P/O)为1 751.1±217.002,表明其繁育系统类型为兼性异交.