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Objective:To investigate the effects of polyethylene glycol 400 （PEG400） on the pharmacokinetics and anti-inflammatory effect of baicalin， and to preliminarily explore the anti-inflammatory effects of baicalin and its main metabolite baicalein 6-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucuronide （B6G） by molecular docking. Method:Rats were randomly divided into two groups with water and PEG400 as the dissolving matrix， and rats were administrated the equal dose of baicalin aqueous solution （baicalin+water group） and baicalin PEG400 solution （baicalin+PEG400 group）. After the plasma samples were processed at different time periods， the concentrations of baicalin and B6G in rat plasma were determined by ultra performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）， and pharmacokinetic parameters were processed by DAS 3.2.2 software. Mice were randomly divided into a blank group （normal saline， 20 mL·kg<sup>-1</sup>）， aspirin group （dose of 0.2 g·kg<sup>-1</sup>）， baicalin/baicalin+PEG400 high and low dose （3.0， 1.5 g·kg<sup>-1</sup>） groups， after continuous administration for 7 days， the mouse ear swelling and foot swelling models were established， and the swelling degree and swelling inhibition rate were calculated. Result:The pharmacokinetic study showed that compared with baicalin+water group， the plasma concentrations of baicalin and B6G increased after administration of baicalin PEG400 solution， and the area under the curve （AUC<sub>0-</sub><italic><sub>t</sub></italic>） increased by 2.36， 1.97 times， and the peak concentration （<italic>C</italic><sub>max</sub>） increased by 2.12， 1.65 times， respectively. The results of mouse ear and foot swelling inflammation models showed that the anti-inflammatory effect was enhanced after intragastric administration of baicalin PEG400 solution. In addition， molecular docking results showed that baicalin and B6G could site bind to multiple target proteins ［tumor necrosis factor （TNF）-<italic>α</italic>， interleukin （IL）-6， IL-1<italic>β</italic>， prostaglandin E<sub>2</sub> （PGE<sub>2</sub>） and nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）］ with higher affinity， which was superior to the positive drug aspirin. Conclusion:PEG400 can increase the plasma concentration of baicalin and its main metabolite B6G， and enhance the anti-inflammatory effect. Baicalin and B6G can form strong hydrogen bonds with various inflammatory factors and of nuclear transcription factors， it is speculated that baicalin and B6G jointly play an anti-inflammatory role.
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The aim of this study was to investigate the effects of polyethylene glycol (PEGs) with different molecular weights (MW: 400, 1 000, 4 000) on the pharmacokinetics of baicalin, and preliminarily analyze its mechanism. Rats were gavaged with baicalin (168 mg·kg-1) + aqueous solution or baicalin + PEGs solution and plasma samples were collected from 0 to 24 h after administration. The concentration of baicalin and its main metabolite baicalein 6-O-β-D-glucuronide (B6G) were determined at different time points by UPLC-MS/MS, and the pharmacokinetic parameters were calculated with DAS 3.0 software. The results showed that PEGs with different molecular weights could effectively increase the AUC0-t of baicalin and B6G, increase the Cmax, and prolong the t1/2, effectively increasing the concentration of baicalin and B6G in vivo. The mechanism may be by promoting the activity of uridine diphosphate glucuronosyl-transferases 1A8 (UGT1A8) and 1A9 (UGT1A9), thereby increasing the transformation rate of baicalin and B6G. The rate of metabolism of B6G was faster than that of baicalin, suggesting that PEGs had a higher affinity for UGT1A8, and PEG400 had the most significant effect. The purpose of this study was to provide a basis for the clinical safe use of baicalin and other flavonoids and the design of new dosage forms with the participation of PEGs. The animal experiment protocol in this study was approved by the Experimental Animal Ethics Committee of Guizhou Medical University.
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Objective::To investigate in vivo and in vitro metabolites of coptisine and their metabolic pathways. Method::SD rats were given coptisine by single gavage (dose of 25 mg·kg-1). Urine and feces from 0 h to 48 h, bile from 0 h to 24 h, and plasma and brain tissue samples at 0.25, 1, 2 h after administration were collected.In vitro metabolism was incubated with rat liver microsomes and intestinal flora.The metabolites were analyzed and identified by the high-resolution HPLC-MS/MS technique.The liquid chromatography separation was carried out on ZORBAX SB-C18 column (4.6 mm×150 mm, 5 μm) with acetonitrile-0.1% formic acid solution as the mobile phase for gradient elution, the flow rate was 1.0 mL·min-1, and column temperature was 25 ℃.The mass spectra were obtained in positive and negative ion mode with electrospray ionization (ESI), the scanning range was m/z 50-1 200.The relative molecular weight was determined according to the quasi-molecular ion peaks.The structures of metabolites were elucidated by comparing the data with literature data, including main ion peaks, UV spectrum and HPLC retention time information. Result::A total of 17 metabolites were identified in each sample, including 11 phase Ⅰ metabolites and 6 phase Ⅱ metabolites.The pathways to these metabolites were hydroxylation, demethylation, dehydrogenation, sulfation and glucuronide conjugation. Conclusion::Coptisine can produce metabolic reaction of phase Ⅰ and phase Ⅱ in rat, and metabolites are predominantly present in urine, and the main metabolic site is liver.Coptisine is poorly absorbed and rarely metabolized in gastrointestinal tract, so it is mostly excreted through feces by prototype.This experiment can provide material basis for the pharmacodynamics and pharmacology of coptisine.
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Objective::To investigate the effect of polyethylene glycol 400 (PEG400) on rat bile excretion of baicalin and its main metabolite [baicalein 6-O-β-D-glucuronide (B6G)], and to analyze its mechanism of action. Method::Rats were randomly divided into baicalin+ water group and baicalin+ PEG400 group, the anesthesia was induced by intraperitoneal injection of 10% chloral hydrate (dose of 4 mL·kg-1) to prepare a rat bile duct intubation model. After the rats were fully awake, rats were given baicalin aqueous solution and baicalin PEG400 solution with dose of 168 mg·kg-1 for baicalin, respectively. And bile was collected from 0 h to 12 h after administration. UPLC-MS/MS was used to determine the concentration of drug excreted through bile at different time periods. Thermo Hypersil GOLD C18 column was used with acetonitrile (A)-0.1% formic acid solution (B) as the mobile phase for gradient elution (0-9 min, 90%-27%B; 9-10 min, 27%-90%B; 10-12 min, 90%B), the flow rate was 0.3 mL·min-1, the column temperature was 30 ℃, the injection volume was 5 μL. The mass spectra were obtained in positive ion mode with electrospray ionization (ESI). The effects of PEG400 on the activities and expressions in rat liver of uridine diphosphate glucuronyltransferase (UGT) 1A8 and UGT1A9 were studied in vitro incubation assay and enzyme linked immunosorbent assay (ELISA). Result::Compared with the baicalin+ water group, in the baicalin+ PEG400 group, the bile cumulative excretions of baicalin and B6G increased by 1.8 times and 2.1 times within 12 h, respectively. PEG400 increased the enzyme activities of UGT1A8 and UGT1A9 by 2.0 times and 1.5 times, and their concentrations in liver were increased by 2.2 times and 1.3 times, respectively. Conclusion::PEG400 can significantly increase the bile excretion of baicalin and its main metabolite B6G by enhancing the activities and expressions of UGT1A8 and UGT1A9, and its promoting effect on bile excretion of B6G is greater than that of baicalin, which provides a basis for the rational clinical application of PEG400 and the design of new dosage forms of flavonoids such as baicalin.
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Objective: To explore the potential Q-markers between crude Scutellaria baicalensis (CSR) and wine-processed S. baicalensis (WSR) based on "components-targets-metabolism" network analysis. Methods: According to the differential components between CSR and WSR, the network relationship of "components-target-metabolomics" was constructed combining network pharmacology and metabolomics. The correlation analysis was then conducted between flavonoids glycosides, and aglycones in S. baicalensis, between differential components and endogenous metabolites to predict the potential quality markers. Results: In this study, combining the results of network pharmacology and metabolomics, baicalin and oroxylin A-7-O-glucuronide were regarded as the quality markers of CSR; Baicalein and wogonin were considered as the quality markers of WSR. Conclusion: It is crucial wine-processed mechanism of S. baicalensis that glutinous rice wine can promote the dissolution and absorption of aglycones. Overall, identification of the differences between Chinese herbal decoction pieces and it processed product, combining analysis of network pharmacology and metabolomics, which provides a demonstration for the investigation of quality markers of Chinese herbal pieces.
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Objective: To explore the network regulation mechanism of Tanreqing Capsule (TRQC) on the treatment of coronavirus disease 2019 (COVID-19). Methods: Potential targets of the 19 major constituents in TRQC were predicted by the Swiss Target Prediction server and TCMSP database. Gene ontology (GO) function enrichment and pathway analysis of the targets were analyzed by Omicsbean analytic system and String 10 database. Finally, Cytoscape 3.6.1 software was used to construct the network pharmacology map. Results: A total of 19 compounds affected 68 pathways such as IL-17 signaling pathway, T cell receptor signaling pathway, arachidonic acid metabolism, cAMP signaling pathway, PI3K-Akt signaling pathway, influenza A, etc, by acting on 163 related targets, which associated with anti-inflammation, immune regulation, antipyresis, eliminating phlegm, relieving cough and asthma, analgesia, antibacterial and antiviral, and sedation. The network of "compound-target-pathway-pharmacological action-efficacy" was also constructed. Conclusion: The major constituents in TRQC, including scutellarin, baicalin, oroxylin-7-O-glucuronide, chrysin-7-O-glucuronide, forsythin, forsythiaside E, forsythiaside D, chlorogenic acid, caffeic acid, isochlorogenic acid A, ursodeoxycholic acid and chenodeoxycholic acid, may interfere with efficacy-related biological processes associated with antipyresis, eliminating phlegm and removing toxin by acting on key protein like TNF, EGFR, NOS3, PTGS2, IL2, GABBR1, MAPK14, ADRB2, REN, VCAM1, ACHE, PTPRC, etc.
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OBJECTIVE：To isolate and purify baicalin metabolites from rat bile ，and to study its effect on the proliferation of liver cancer HepG 2 cells. METHODS ：Totally of 6 SD rats were collected ，anesthetized and then intubated with bile ducts. They were given baicalin （168 mg/kg）intragastrically after the rats were awake as well as the bile sample 1 was collected during 24 h after intragastric administration. After processed ，bile sample 1 was preliminarily analyzed by HPLC. Another 5 SD rats were anesthetized and intubated in the same way as above ，and then given baicalin intragastrically （400 mg/kg）. The bile sample 2 was collected during 48 h after intragastric administration. After processed ，the bile sample 2 was isolated and purified by semi-preparative HPLC. The isolated metabolites were identified by using UV ，IR，MS and NMR ，as well as based on physical and chemical properties. MTT method combined with high content cell imaging analysis system was used to study the effect of the metabolites on the proliferation of HepG 2 cells. RESULTS ：Baicalin was mainly metabolized into metabolite 1 and metabolite 2 through bile. After isolation and purification ， they were identified as oroxylin A- 7-O-β-D-glucuronide and 2726719792@qq.com baicalein 6-O-β-D-glucuronide. The results of MTT assay showed that the metabolite 2 had a significant inhibitory effect 分析。E-mail：gaoxl@gmc.edu.cn on the proliferation of HepG 2 cells，and its IC 50 was 90 µg/mL；the results of high content cell imaging analysis showed that at a certain concentration metabolite 2 may inhibit proliferation by changing the mitochondrial distribution and membrane permeability of HepG 2 cells（studies on the pharmacological activities of metabolism 1 had been reported ，it was skipped in this study ）. CONCLUSIONS ：In this study ，two baicalin bile metabolites were successfully isolated and identified ，of which baicalein 6-O-β-D-glucuronide has a significant inhibitory effect on the proliferation of HepG 2 cells.
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The quality evaluation method for standard decoction of Chinese herbal slices is the basis for the quality evaluation of granules and preparations of classical formula(decoction)of traditional Chinese medicine. This study aimed to establish a method for the determination of quercetin-3-O-glucuronic acid in Nelumbinis Folium(NF)and its standard decoction, so as to provide reference for the quality control of NF and its standard decoction. Fifteen batches of representative NF were collected to prepare standard decoction, and the parameters of dry extract rate, transfer rate of index component, and pH value were calculated. HPLC was used to establish the content determination method for quercetin-3-O-glucuronic acid in NF and its standard decoction. The concentration range of quercetin-3-O-glucuronic acid in the standard decoction of NF was 1.09-3.06 g·L~(-1), while the concentration range of nuciferine was 0.01-0.17 g·L~(-1). The average extraction rate of NF standard decoction was(14.4±2.6)%, the average transfer rate of quercetin-3-O-glucuronic acid was(70.7±18.6)%, and the average transfer rate of nuciferine was(9.6±5.4)%. Compared with Nuciferine, quercetin-3-O-glucuronic acid had a high content and stable transfer rate in standard decoction, and was recommended to be the quality control marker for NF and its standard decoction. This paper establishes a quality evaluation method for NF standard decoction, and can provide reference for the quality control of all preparations derived from NF and its decoction.
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Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Flowers/chemistry , Medicine, Chinese Traditional , Nelumbo/chemistry , Quality ControlABSTRACT
Objective: To establish multiwavelength HPLC fingerprint of Aurea helianthus from different batches, and combine quantitative analysis, similarity evaluation, cluster analysis, and principal component analysis to evaluate the quality of A. helianthus. Methods: The chromatographic column was Phenomenex Kinetex C18 (250 mm × 4.6 mm, 5 μm). The mobile phase was composed of 0.08% phosphoric acid water (A) and acetonitrile (B) in gradient elution at a flow rate of 1.0 mL/min, the detection wavelength was set at 260 nm for protocatechuic acid during 0-8 min, 324 nm for caffeic acid during 8-15 min, 360 nm for rutin, hyperin, isoquercitrin, gossypetin-8-O-β-D-glucuronide, myricetin, quercetin-3’-O-glucoside, and quercetin during 15-60 min. The column temperature was set at 30 oC. And the HPLC fingerprint of A. helianthus was established by the similarity evaluation system for chromatographic fingerprint of TCM (Version 2004A) and SPSS19.0, which was used for similarity evaluation, cluster analysis, and principal component analysis. Results: A total of 25 common peaks were confirmed of A. helianthus HPLC fingerprint, and nine peaks were identified which were determined. The similarity of 16 batches of samples was between 0.879 and 0.983; The results of cluster analysis showed that A. helianthus was clustered into two groups, indicating that there were differences in the similarity; Ranked the quality of A. helianthus based on the main component composite score. Conclusion: The method is simple and accurate, which can be used for the comprehensive quality evaluation research of the medicinal materials of A. helianthus.
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Objective: To study the antioxidant chemical constituents and antioxidant activity of Patrinia villosa. Methods: The 70% ethanol-water extract of the herb was separated by silica gel column chromatography, ODS column chromatography and sephadex column chromatography. Then, the compound were further purified and extracted by semi-preparative HPLC. Their structures were elucidated by physiochemical property and spectral analysis. DPPH and ABTS methods were used to determine the antioxidant bioactivities of the isolated compounds. Results: A total of ten compounds were isolated and synthesized, including chlorogenic acid butyl ester (1), 3,4-di-O-caffeoyl quinic acid methyl ester (2), luteolin-7-O-rutinoside (3), 1β-O-β-D-glucopyranosy- 15-O-(p-hydroxylphenylacetate)-5α,6βH-eudesma-3,11(13)-dien-12,6α-olide (4), 3,4-di-O-caffeoyl quinic acid ethyl ester (5), 4,5-di-O-caffeoyl quinic acid methyl ester (6), 4,5-di-O-caffeoyl quinic acid n-butyl ester (7), luteolin-7-O-β-D-glucuronide methyl ester (8), luteolin-7-O-β-D-glucuronide ethyl ester (9), and apigenin-7-O-β-D-glucuronide methyl ester (10). The DPPH radical scavenging IC50 of compounds 3, 8, and 9 were (23.95 ± 0.71), (73.09 ± 0.33), and (25.06 ± 0.65) μmol/L, respectively. The ABTS radical scavenging IC50 was (7.13 ± 0.07), (11.48 ± 0.21), (5.15 ± 0.08) mol/L, respectively. Conclusion: Eight compounds except compounds 3 and 8 are obtained from this species for the first time. Compounds 3, 8, and 9 had significant antioxidant activity.
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Managing the dysregulated host response to infection remains a major challenge in sepsis care. Chinese treatment guideline recommends adding XueBiJing, a five-herb medicine, to antibiotic-based sepsis care. Although adding XueBiJing further reduced 28-day mortality modulating the host response, pharmacokinetic herb-drug interaction is a widely recognized issue that needs to be studied. Building on our earlier systematic chemical and human pharmacokinetic investigations of XueBiJing, we evaluated the degree of pharmacokinetic compatibility for XueBiJing/antibiotic combination based on mechanistic evidence of interaction risk. Considering both XueBiJing‒antibiotic and antibiotic‒XueBiJing interaction potential, we integrated informatics-based approach with experimental approach and developed a compound pair-based method for data processing. To reflect clinical reality, we selected for study XueBiJing compounds bioavailable for drug interactions and 45 antibiotics commonly used in sepsis care in China. Based on the data of interacting with drug metabolizing enzymes and transporters, no XueBiJing compound could pair, as perpetrator, with the antibiotics. Although some antibiotics could, due to their inhibition of uridine 5'-diphosphoglucuronosyltransferase 2B15, organic anion transporters 1/2 and/or organic anion-transporting polypeptide 1B3, pair with senkyunolide I, tanshinol and salvianolic acid B, the potential interactions (resulting in increased exposure) are likely desirable due to these XueBiJing compounds' low baseline exposure levels. Inhibition of aldehyde dehydrogenase by 7 antibiotics probably results in undesirable reduction of exposure to protocatechuic acid from XueBiJing. Collectively, XueBiJing/antibiotic combination exhibited a high degree of pharmacokinetic compatibility at clinically relevant doses. The methodology developed can be applied to investigate other drug combinations.
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Objective: By investigating the material composition, anti-oxidant and hypoglycemic activity of acetonitrile-water extracts of Origanum vulgare leaf (OL) in different harvest time, to acquire the scientific data for the utilization of OL. Methods: OL was extracted with acetonitrile and water (1: 1). The contents of total phenols and total flavonoid were measured by Foline-Phenol reagent method and AlCl3 colorimetry. The main compositional analysis was performed using LC-QTOF-MS technology. Meanwhile, the anti-oxidation of OL extracts in different harvest time was evaluated by total anti-oxidant capacity assay kit with ABTS method and ferric reducing anti-oxidant potential assay (FRAP). The methods of 2-NBDG glucose uptake and α-glycosides inhibition were applied to evaluate hypoglycemic activity of OL extracts. Results: The six main components in OL extract were identified to be origanoside I, luteolin 7-O-glucuronide, apigenin 7-O-glucuronide, rosmarinic acid, lithospermic acid and origanoside I derivative. The contents of total phenols and the total flavonoids were the highest in OL extracts harvested in July and September, respectively. There was good in vitro anti-oxidant and hypoglycemic activity for OL extracts harvested in different times. Among them, the best anti-oxidant activity was observed in OL extracts harvested in July, while the best hypoglycemic activity was observed in OL extracts harvested in October. Conclusion: OL has potential anti-oxidant and hypoglycemic activity, which is affected by the harvest time.
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Objective To study the chemical constituents of Gendarussa vulgaris. Methods The chemical constituents were isolated and purified with silica column chromatography and gel chromatography, etc. Their structures were identified by physico- chemical properties and various spectroscopic methods including NMR spectrum, MS, UV, etc. Results A total of 16 compounds were isolated and elucidated as daucosterol (1), 6″-O-acetylisavitexin (2), 9,10-dihydroxy-4,7-megastigmadien-3-one (3), 22E,24R-ergosta- 7,22-diene-3β,5α,6β,9α-tetraol (4), isorhamnetin (5), quercetin (6), eleutheroside E (7), gusanlung A (8), gusanlung B (9), betulin (10), isovitexin-2″-O-rhamnoside (11), isovitexin (12), genkwanin (13), apigenin (14), quercetin 3-O-β-D-glucuronide (15), and 2-(4-hydroxy-3-methoxyphenyl)-3-(2-hydroxy-5-methoxyphe-nyl)-3-oxo-1-propanol (16). Conclusion Compounds 3, 5, 11, 12 and 15 are isolated from G. vulgaris for the first time. Compound 2, 4, 13, and 16 are isolated from the plant of genus for the first time.
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Objective To study the chemical constituents and bioactivities of leaves of Torreya grandis. Methods The chemical constituents were isolated by MCI-Gel CHP-20, Diaion HP-20, Toyopearl HW-40, Sephadex LH-20, RP-18 and silica gel column chromatographic methods. Their structures were identified on the basis of physicochemical and spectroscopic analysis. The cytotoxic activity and antitumor activity of compounds 1-4 were investigated by lethal-to-prawn larva bioactivity determining method and MTT assay. Results Ten compounds were isolated from EtOAc extract and n-BuOH of leaves of T. grandis. Their structures were identified as torreyagrandate (1), hinokiol (2), 4-epiagathadial (3), 3,4-dihydroxybenzoic acid 3-O-β-D-glucoside (4), dehydroabietic acid (5), trans-communic acid (6), cis-communic acid (7), (2-methoxy-1,4-phenylene) dimethanol (8), pinoresinol (9), and β-sitosterol (10). The bioactivity experiment indicated that compounds 1-4 possessed certain cytotoxic activity towards brine shrimp, and LC50 value were 7.7, 8.0, 8.8, and 4.2 μg/mL, respectively. In addition, it was found that compound 4 presented remarkable inhibitory effect on two kinds of cells of human liver cancer cells of Huh7 (67%) and HepG2 (69%) at a dose of 10 μg/mL. Conclusion All compounds are isolated from leaves of T. grandis for the first time expect compound 10. Compounds 1-4 exhibite certain cytotoxic activity, and compound 4 displays stronger antitumor activity towards Huh7 and HepG2 liver cancer cells.
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Flavonol glycoside is in clinical trials for treatment of hyperlipidemia. An accurate and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of flavonol glycoside (M0), aglycone (M1) and glucuronide conjugate (M2) in rat plasma. d6-Flavonol glycoside was used as internal standard (IS). After extraction from the plasma by protein precipitation, the analytes and internal standard were separated on a XDB C18 column (50 mm×4.6 mm, 1.8 μm) using a gradient elution procedure. The mobile phase consisted of methanol and water (0.2% formic acid) at a flow rate of 0.6 mL·min−1. The total run time was 4.5 min. Positive electrospray ionization was performed using multiple reaction monitoring (MRM) with transitions of m/z 461.3 → m/z 299.1 for M0, m/z 299.1 → m/z 283.1 for M1, m/z 475.0 → m/z 299.1 for M2, and m/z 467.3 → m/z 305.1 for d6-flavonol glycoside. The method was validated and successfully applied to the pharmacokinetics study of flavonol glycoside in SD rats which were given flavonol glycoside (30 mg·kg−1) by gavage. The Cmaxof M0 is (341 ±106) ng·mL−1 and AUC0−t is (1 960 ±725) h·ng·mL−1, while the Cmaxof M2 is (1 720 ±843) ng·mL−1and AUC0−t is (8 510 ±2 920) h·ng·mL−1. The results suggest that flavonol glycoside existed mainly in the form of M0 and M2 in rats. After flavonol glycoside being hydrolyzed by the intestinal flora, it was absorbed in the form of aglycone and further metabolized to M2 after the first-pass effect. In this paper, the main metabolites of flavonol glycoside in rat plasma were determined for the first time, which provided a basis for the design of clinical pharmacokinetic experiment.
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OBJECTIVE To investigate the protective effect and mechanisms of luteolin-7-O-β-d-glucuronide (LGU) on oxygen glucose deprivation (OGD)-induced H9C2 cardiomyocytes injury. METH-ODS The protective effect of LGU on OGD-induced H9C2 cardiomyocytes death were investigated by MTT assay. The microfilament change of H9C2 cardiomyocytes was detected by phalloidin staining and the lactate dehydrogenase (LDH) leakage rate was also detected by LDH kit. In order to explore the possible mechanisms of LGU, ATP content, intracellular Ca2+fluorescent intensity and concentra-tion, mitochondrial membrane potential (MMP)and the expressions of apoptosis-related proteins were detected by ATP kit,CLSM(Fluo-3/AM probe),Ca2+kit,CLSM(JC-1 probe)and western blotting meth-od, respectively. RESULTS The inhibition of H9C2 cardiomyocyte survival rate inducedby OGD was improvedby pretreated with LGU in a concentrationdependent manner. The microfilaments injury as well as the increase of LDH leakage rate were also improvedby pretreated with LGU.The ATP content was significantly decreased,intracellular Ca2+fluorescent intensity and concentration were significantly increased and the MMP was significantly decreased 4 hafter OGD. LGU significantly reversed the de-crease of intracellular ATP content,the increase of Ca2+fluorescent intensity and concentration and the decrease of MMP.The release of cytochrome C,the expressionsof caspase-9 and caspase-3 in H9C2 cardiomyocytes were increased 16 h after OGD.LGUsignificantly inhibited the changes of these apop-tosis-related proteins. CONCLUSION LGU has a significant protective effect against OGD-induced H9C2 cardiomyocytes injury through inhibiting calcium overload,increasing ATP content,improving mi-tochondrial function and inhibiting apoptosis.
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OBJECTIVE To investigate the neuroprotective effect and possible mechanisms of lute-olin-7-O-β-D-glucuronide (LGU) against focalcerebral ischemic injury. METHODS The focal cerebral ischemic injury model was established by middle cerebral artery occlusion (MCAO). Male Sprague Dawley rats were randomly divided into sham group,model group(MCAO),LGU group(0.24,0.72 and 2.16 mg·kg-1)and positive control group(Edaravone at 5 mg·kg-1).LGU was injected intravenously 30 min after MCAO.Neurological severity score,infarct volume and brain water content were detected 24 h after MCAO and the levels of Na+-K+ATPase,Ca2+ATPase,TNF-α and IL-1β were detected to explore the possible mechanisms.For the therapeutic time window test,LGU(0.72 mg·kg-1)was injected intrave-nously 0.5, 2, 4, 6, 8, 10 and 12 h respectively after MCAO. To evaluate motion behavior, LGU were injected intravenously 30 min after MCAO and once per day during detection period. The changes of motor coordination were detected by rotating rod method and grip strength analysis, and the changes of gaits were detected using DigiGait Imaging System. RESULTS LGU improved the neurological severity score, infarct volume ratio and brain water content. The therapeutic time window of LGU for cerebral infarction and brain edema was at least 6 h and for neurological dysfunction was 12 h.LGU also prolonged the latency on rotarod, increased the forelimb tension and improved 8 gait parameters, including stance duration,stride length,stance width,paw area,paw area variability,gait symmetry,ataxia coefficient and tau propulsion.Furthermore,LGU increased Na+-K+-ATPase and Ca2+-ATPase levels in the cortex and hippocampus in the ischemic side,reduced the levels of TNF-α and IL-1β in the serum. CONCLUSION LGU has a significant neuroprotective effect against cerebral ischemic injury via improving energy metabolism and reducing inflammation.
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Abstract Activity of hepatic metabolic enzymes of glucuronidation and sulfation of 4-nitrophenol (PNP) and biliary excretion of its glucuronide (PNP-G) and sulfate (PNP-S) conjugates have been investigated in control and streptozotocin (STZ)-induced diabetic rats. 500 µM PNP solution was luminally perfused in a cannulated jejunal loop for 90 minutes. It was found that biliary excretion of PNP-G was significantly decreased in the diabetic rats. This effect of STZ could be completely reversed by administration of rapid-acting insulin. Activity of hepatic UDP-glucuronyltransferase and ß-glucuronidase was also depressed by the STZ pretreatment. Administration of insulin antagonized the inhibitory action of STZ on UDP-glucuronyltransferase, but the reduced activity of ß-glucuronidase was not reversed. Biliary excretion of PNP-S was also depressed in the diabetic rats. Whereas, different effects of insulin administration were observed. Namely, the lower biliary excretion rate of PNP-S was not changed after administration of insulin. Activity of the sulfotransferase and the arylsulfatase enzymes was not altered either by STZ pretreatment or by insulin administration. Biliary excretion of PNP was also significantly depressed by STZ and this depression was not changed after insulin administration. The results call attention to hepatobiliary circulation of low molecular weight xenobiotics and their glucuronide and sulfate conjugates
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Animals , Male , Rats , Diabetes Mellitus, Experimental/chemically induced , Hepatobiliary Elimination , Streptozocin , Hepatobiliary Elimination/immunologyABSTRACT
Objective: To study the metabolites from rats urine after ig administration of calycosin-7-O-β-D-glucopyranoside. Methods: The constituents were isolated and purified by preparative liquid chromatographic technique, and the structures were identified by spectroscopic analyses including ESI-MS, NMR, and 2D-NMR. Results: Five metabolites were isolated from rats urine after ig administration of calycosin-7-O-β-D-glucopyranoside. They were identified as calycosin (M1), 3', 4', 7-trihydroxyisoflavone (M2), daidzein (M3), calycosin-3'-O-β-D-glucuronide (M4), and calycosin-3'-O-β-D-glucuronide methyl ester (M5). Conclusion: M5 is a new compound.
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Objective To develop a sensitive and reproducible liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to evaluate the pharmacokinetic behavior of berberrubine (BRB) and its glucuronide (BRBG) in rats. Methods BRB, BRBG and tetrahydroberberine (THB, internal standard) were isolated by liquid-liquid extraction in rat biological samples. Chromatographic separation was achieved on an Agilent Zorbax Eclipse Plus C18 (2.1 mm × 50 mm, 3.5-Micron) with a gradient mobile phases primarily containing acetonitrile, water with 0.1% formic acid and 5 mm ammonium acetate. The analytes were monitored by MS/MS in positive electrospray ionization mode. Herein, the feasibility of new developed method was validated with respect to specificity, linearity, precision, accuracy, stability, extraction efficiency and matrix effect. The appropriate method was used for the pharmacokinetic study in rats. Results The new developed method could be applied to the pharmacokinetic study of BRB in rats. BRB and BRBG showed good linearity over the ranges of 2-1000 ng/mL and 5-2000 ng/mL, respectively, and precision was no more than 15%. The accuracy, specificity and stability could be acceptable. Conclusion The new method is sensitive and reproducible. In pharmacokinetic study, BRB showed nonlinear elimination property. Meanwhile, BRB was rapidly absorbed and widely distributed in various tissues with the highest exposure of BRB in kidney and liver. The absolute bioavailability of BRB was determined to be 8.2% and at the dose of 40 mg/kg, a total of 44% BRB was excreted in urine and feces.