Carthami Flos, as a traditional blood-activating and stasis-resolving drug, possesses anti-tumor, anti-inflammatory, and immunomodulatory pharmacological activities. Flavonoid glycosides are the main bioactive components in Carthamus tinctorius. Glycosyltransferase deserves to be studied in depth as a downstream modification enzyme in the biosynthesis of active glycoside compounds. This study reported a flavonoid glycosyltransferase CtUGT49 from C. tinctorius based on the transcriptome data, followed by bioinformatic analysis and the investigation of enzymatic properties. The open reading frame(ORF) of the gene was 1 416 bp, encoding 471 amino acid residues with the molecular weight of about 52 kDa. Phylogenetic analysis showed that CtUGT49 belonged to the UGT73 family. According to in vitro enzymatic results, CtUGT49 could catalyze naringenin chalcone to the prunin and choerospondin, and catalyze phloretin to phlorizin and trilobatin, exhibiting good substrate versatility. After the recombinant protein CtUGT49 was obtained by hetero-logous expression and purification, the enzymatic properties of CtUGT49 catalyzing the formation of prunin from naringenin chalcone were investigated. The results showed that the optimal pH value for CtUGT49 catalysis was 7.0, the optimal temperature was 37 ℃, and the highest substrate conversion rate was achieved after 8 h of reaction. The results of enzymatic kinetic parameters showed that the K_m value was 209.90 μmol·L~(-1) and k_(cat) was 48.36 s~(-1) calculated with the method of Michaelis-Menten plot. The discovery of the novel glycosyltransferase CtUGT49 is important for enriching the library of glycosylation tool enzymes and provides a basis for analyzing the glycosylation process of flavonoid glycosides in C. tinctorius.
Carthamus tinctorius/chemistry* ; Phylogeny ; Flavonoids/analysis* ; Glycosides/analysis* ; Glycosyltransferases/genetics* ; Anti-Inflammatory Agents ; Chalcones

In order to solve the problem of weak correlation between quality control components and efficacy of Glycyrrhizae Radix et Rhizoma, this study detected the interaction between small molecular chemical components of Glycyrrhizae Radix et Rhizoma and total proteins of various organs of mice by fluorescence quenching method to screen potential active components. The 27 chemical components in Glycyrrhizae Radix et Rhizoma were detected by HPLC and their deletion rates in 34 batches of Glycyrrhizae Radix et Rhizoma were calculated. Combined with the principle of component effectiveness and measurability, the potential quality markers(Q-markers) of Glycyrrhizae Radix et Rhizoma were screened. RAW264.7 macrophage injury model was induced by microplastics. The cell viability and nitric oxide content were detected by CCK-8 and Griess methods. The levels of inflammatory factors(TNF-α, IL-1β, IL-6, CRP) and oxidative stress markers(SOD, MDA, GSH) were detected by the ELISA method to verify the activity of Q-markers. It was found that the interaction strength between different chemical components and organ proteins in Glycyrrhizae Radix et Rhizoma was different, reflecting different organ selectivity and 18 active components were screened out. Combined with the signal-to-noise ratio of the HPLC chromatographic peaks and between-run stability of the components, seven chemical components such as liquiritin apioside, liquiritin, isoliquiritin apioside, isoliquiritin, liquiritigenin, isoliquiritigenin and ammonium glycyrrhizinate were finally screened as potential Q-markers of Glycyrrhizae Radix et Rhizoma. In vitro experiments showed that Q-markers of Glycyrrhizae Radix et Rhizoma could dose-dependently alleviate RAW264.7 cell damage induced by microplastics, inhibit the secretion of inflammatory factors, and reduce oxidative stress. Under the same total dose, the combination of various chemical components could synergistically enhance anti-inflammatory and antioxidant effects compared with the single use. This study identified Q-markers related to the anti-inflammatory and antioxidant effects of Glycyrrhizae Radix et Rhizoma, which can provide a reference for improving the quality control standards of Glycyrrhizae Radix et Rhizoma.
Mice ; Animals ; Antioxidants/analysis* ; Microplastics/analysis* ; Plastics/analysis* ; Rhizome/chemistry* ; Drugs, Chinese Herbal/analysis* ; Glycyrrhiza/chemistry* ; Anti-Inflammatory Agents/analysis*

OBJECTIVETo study the anti-inflammatory and antioxidant effects of Petasites tricholobus and its phenolic components.

METHODPhenolic compounds were separated purified by column chromatographic methods such as macroporous resin D-101, silica gel, ODS, MCI GEL CHP 20P, Sephadex LH-20. Their structures were identified on the basis of physicochemical property and multiple spectral data.

RESULTNineteen phenolic compounds were separated from 95% ethanol extracts from P. tricholobus Franch. and identified as sulfonated benzyl glucoside (1), 3-(4beta-D-glucopyranosyloxy-3, 5-dimethoxy) -phenyl-2E-propenol (2), dihydrosyringin (3), tangshenosides II (4), 4-hydroxy-2,6-dimethoxyphenol-1-O-beta-D-glucopyranoside (5), 4-hydroxymethyl-2, 6-dimethoxyphenyl-1-O-beta-D-glucopyranoside (6), arbutin (7), rutin (8), kaempferol-3-O-alpha-L-rhamnopyranosyl-(1 --> 6)-beta-D-glucopyranoside (9), quercetin-3-O-beta-D-glucopyranoside (10), kaempferol-3-O-beta-D-glucopyranoside (11), afzelin (12), petasiphenol (13), caffeic acid (14), chlorogenic acid (15), 2-hydroxy-5-acetylbenzoic acid (16), p-hydroxy-benzoic acid (17), protocatechuic aldehyde (18) , and p-hydroxy-phenylpropionic acid (19).

CONCLUSIONAbove result shows that phenolic compounds contained in P. tricholobus mainly include simple phenols, phenolic glycosides, coffee acid and flavonoid glycosides. Among them, compound 1 was separated from the composite family for the first time; compounds 2-7, 9, 11, 12, 16, 19 were separated from the genus Petasites for the first time, and the others were separated from the plant for the first time. These compounds have been proved to have pharmacological effects such as anti-inflammation, antibiosis, antioxidantion.

Anti-Inflammatory Agents ; analysis ; isolation & purification ; Antioxidants ; analysis ; isolation & purification ; Asteraceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Phenols ; analysis ; isolation & purification

UPLC-ESI-Orbitrap-MS/MS was used to analyze,identify and attribute the chemical constituents in Pudilan Antiphlogistic Oral Liquid. The analysis was performed on an Agilent Eclipse XDB-C18(4.6 mm × 150 mm,3.5 μm) with a gradient mobile phase of methanol-0.1% formic solution system at the flow rate of 0.5 m L·min-1. The sample volume was 2 μL. The column temperature was30 ℃. The high-resolution orbitrap mass spectrometry was used as detector,with electrospray ion source in both positive and negative models,and the MS scanning ranged between m/z 50 and 2 000. Based on the analysis of mass spectrometry and literature reports,79 compounds were confirmed,including 30 alkaloids,28 organic acids,18 flavonoids and 3 coumarins. Finally,39 compounds,such as rutin,esculetin,gallic acid,caffeic acid,cichoric acid,were identified from Taraxacum mongolicum; 11 compounds,such as baicalin,baicalein,apigenin,chrysin,oroxylin A,were identified from Scutellaria baicalensis; 13 compounds,such as arginine,proline,hypoxanthine,epigoitrin,indirubin,were identified from Isatis indigotica; and 18 compounds,such as dehydrocheilanthifoline,oxysanguinarine,corynoline,protopine,spallidamine,were identified from Corydalis bungeana. After the analysis of chemical model and attribution,the contents of some compounds were high in Pudilan Antiphlogistic Oral Liquid,such as baicalin,wogonoside,baicalein,wogonin,apigenin,chrysin,skullcapflavonⅡ,oroxylin A,cichoric acid,chlorogenic acid,caffeic acid,esculetin,dehydrocheilanthifoline,dihydrosanguinarine,protopine,corynoline and indirubin. The established method is simple,accurate,rapid,sensitive and reproducible,and thus suitable for the qualitative identification and quantitative determination of Pudilan Antiphlogistic Oral Liquid,which lays a foundation for the systematic quality control and the establishment of whole-course traceability system of active ingredients.
Anti-Inflammatory Agents ; analysis ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Phytochemicals ; analysis ; Tandem Mass Spectrometry

Plants have been used as a source for food material and natural remedies for the treatment of vast range of diseases. Nature provides us remedies for the treatment of various types of disorders ranging from simple ailments to complicated diseases. Plants are known to possess different pharmacological activities due to the presence of various phytoconstituents. Flavonoids are one of the main active phytoconstituents found in fruits, vegetables, wines, tea and cocoa. Flavonoids exhibit various pharmacological activities such as antioxidant, anti-inflammatory, anti-allergic, antibacterial, oestrogenic, cytotoxic antitumoural, hepatoprotective, antithrombotic and antiviral activity. Diosmetin (3', 5, 7-trihydroxy-4'-methoxyflavone), the aglycone part of the flavonoid glycosides diosmin occurs naturally in citrus fruit. Although it is found in herbal medicines and plays an important role in the treatment of various ailments, only limited scientific researches have been conducted. The aim of this review is to collect all available scientific literature published on diosmetin and combine it into this paper. This review contains an overview of pharmacological activities, isolation techniques and analytical techniques for diosmetin. Thus, valuable information provided in the present review will help researchers in developing alternative methods for the treatment of diseases from diosmetin.
Anti-Bacterial Agents ; analysis ; chemistry ; pharmacokinetics ; pharmacology ; Anti-Inflammatory Agents ; analysis ; chemistry ; pharmacokinetics ; pharmacology ; Antineoplastic Agents ; analysis ; chemistry ; pharmacokinetics ; pharmacology ; Antioxidants ; analysis ; chemistry ; pharmacokinetics ; pharmacology ; Flavonoids ; analysis ; chemistry ; pharmacokinetics ; pharmacology ; Humans

OBJECTIVETo investigate the effect of Angelicae Pubescentis Radix (APR) on the activity of endocannabinoid hydrolase and N-acylethanolamine-hydrolyzing acid amidase (NAAA), and to demonstrate the mechanism of anti-inflammatory effect of APR by in vitro lipopolysaccharide (LPS)-induced inflammation model.

METHODAPR essential oil was extracted by steam distillation, and the chemical components were identified by GC-MS. Enzymatic activity was performed by using recombinant NAAA-overexpressing protein and detected by LC-MS. Lipids were extracted by methonal/chloroform mixure and analyzed by LC-MS. mRNA and protein expression levels of proinflammatory genes were examined by Real time-PCR and ELISA assay kit, respectively. The content of nitro oxide (NO) was detected by Griess reaction.

RESULTTwenty active components were identified from APR essential oil which inhibited NAAA activity in a dose-dependent manner. On the LPS-induced RAW264.7 cells, APR essential oil reversed LPS-suppressed N-palmitoylethanolamide (PEA) contents in a dose-dependent manner and reduced LPS-induced proinflammatory genes, TNF-alpha and IL-6. Moreover, APR essential oil reduced the mRNA expression of iNOS, subsequently reduced the release of NO, a classic inflammatory marker.

CONCLUSIONThe research demonstrated that the effect of APR on inflammation is mediated by the inhibition of NAAA activity, which increase the cellular endobioactor PEA levels and decrease proinflammatory factor. The results suggest that APR can serve as a nature NAAA inhibitor.

Amidohydrolases ; antagonists & inhibitors ; Angelica ; chemistry ; Animals ; Anti-Inflammatory Agents ; pharmacology ; Enzyme Inhibitors ; pharmacology ; Lipopolysaccharides ; pharmacology ; Mice ; Oils, Volatile ; analysis ; pharmacology

OBJECTIVETo assess the analgesic and anti-inflammatory effects of Tongjingbao optimal formula and analyze its active components.

METHODAnimals were divided into the model group, the Tongjingbao granule group and the Tongjingbao optimal formula group. The mice dysmenorrhea model was induced by oxytocin, and their content of blood calcium and MDA, NO, PGE2 in uterus were determined to assess the analgesic and anti-inflammatory effects of different components in Tongjingbao optimal formula and their impacts.

RESULTAll components of Tongjingbao optimal formula could extend the dysmenorrhea incubation period of mice with dysmenorrhea, reduce their average writhing time, increase the writhing inhibition rate, lessen the content of blood calcium and MDA, PGE2 in uterus, and enhance the content of NO in uterus.

CONCLUSIONAll components of Tongjingbao optimal formula have the analgesic and anti-inflammatory effects, and different components show a synergistic effect in treating dysmenorrheal in many links.

Analgesics ; administration & dosage ; analysis ; Animals ; Anti-Inflammatory Agents ; administration & dosage ; analysis ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; Dysmenorrhea ; drug therapy ; Female ; Humans

The paper is to report the establishment of three methods for determination of methyl salicylate-2-O-beta-D-galactopyranoside (1-4)-beta-D-glucopyranoside (MSG) by HPLC, UV or potentiometric titration. The results determined by the three methods turned out to be of no significant difference (P>0.05). The method was chosen according to MSG difference test demands.
Anti-Inflammatory Agents ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Glycosides ; analysis ; chemistry ; Molecular Structure ; Potentiometry ; methods ; Reproducibility of Results ; Salicylates ; analysis ; chemistry ; Sensitivity and Specificity ; Spectrophotometry, Ultraviolet ; methods

PURPOSE: The purpose of this study was to determine the impact of aspirin, nonsteroidal anti-inflammatory drugs (NSAIDs), statin, and cyclooxygenase 2 (COX-2) inhibitor on the development of kidney, prostate, and urothelial cancers by analyzing the Korean National Health Insurance Service–National Sample Cohort (NHIS-NSC) database. MATERIALS AND METHODS: Among a representative sample cohort of 1,025,340 participants in NHIS-NSC database in 2002, we extracted data of 799,850 individuals who visited the hospital more than once, and finally included 321,122 individuals aged 40 and older. Following a 1-year washout period between 2002 and 2003, we analyzed 143,870 (male), 320,861 and 320,613 individuals for evaluating the risk of prostate cancer, kidney cancer and urothelial cancer developments, respectively, during 10-year follow-up periods between 2004 and 2013. The medication group consisted of patients prescribed these drugs more than 60% of the time in 2003. To adjustfor various parameters of the patients, a multivariate Cox regression model was adopted. RESULTS: During 10-year follow-up periods between 2004 and 2013, 9,627 (6.7%), 1,107 (0.4%), and 2,121 (0.7%) patients were diagnosed with prostate cancer, kidney cancer, and urothelial cancer, respectively. Notably, multivariate analyses revealed that NSAIDs significantly increased the risk of prostate cancer (hazard ratio [HR], 1.35). Also, it was found that aspirin (HR, 1.28) and statin (HR, 1.55) elevated the risk of kidney cancer. No drugs were associated with the risk of urothelial cancer. CONCLUSION: In sum, our study provides the valuable information for the impact of aspirin, NSAID, statin, and COX-2 inhibitor on the risk of prostate, kidney, and urothelial cancer development and its survival outcomes.
Anti-Inflammatory Agents ; Anti-Inflammatory Agents, Non-Steroidal ; Aspirin* ; Cohort Studies ; Cyclooxygenase 2 ; Follow-Up Studies* ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors* ; Kidney ; Kidney Neoplasms ; Korea* ; Multivariate Analysis ; National Health Programs ; Prostate ; Prostatic Neoplasms

Dexamethasone sodium phosphate (DSP) is increasingly used in the treatment of ocular inflammatory diseases by systemic, periocular, and recently, intravitreal injection. We have developed a method for the determination of vitreous levels of DSP by reverse phase HPLC. In this method, co-elution of vitreous proteins with DSP is resolved by a prior sample clean-up procedure using Waters Sep-Pak C18 cartridge; the protein is separated and eluted with water while DSP, paraben and prednisone are eluted with methanol. DSP in the resulting sample is then separated by reverse phase HPLC and quantified by UV absorption at 254 nm. The recovery of DSP through the sample clean-up is 68.9 +/- 3.0%. DSP quantitation is linear from 0.1 mg to 1.0 mg per 1.0 ml vitreous. This method provides a simple, sensitive and reliable technique for determining the vitreous levels of DSP.
Animals ; Anti-Inflammatory Agents/*analysis ; Chromatography, High Pressure Liquid/*methods ; Dexamethasone/*analogs & derivatives/analysis ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity ; Vitreous Body/*chemistry