As its tuberous alkaloids induce valuable pharmacological effects,Pinellia ternata has considerable clinical value.However,its production currently fails to meet its demand.In vitro microtuber culture,combined with sa...As its tuberous alkaloids induce valuable pharmacological effects,Pinellia ternata has considerable clinical value.However,its production currently fails to meet its demand.In vitro microtuber culture,combined with salicylic acid(SA)elicitation,may provide an effective alternative to traditional field production.In this study,an in vitro P.ternata microtuber induction system was developed and used to evaluate SA-induced elicitation of alkaloid accumulation.The quality of in vitro microtubers was assessed by total alkaloid measurement,a cytotoxicity assay,and transcriptomic analysis.With or without SA treatment,P.ternata microtuber induction was achieved within 60 d using petiole-derived explants,with a microtuber proliferation rate of approximately 17 microtubers per explant.The total alkaloid content of in vitro microtubers elicited with 100μM SA was equivalent to that of field-grown tubers,while those not treated with SA contained lower alkaloid content.The cytotoxicity assay showed preliminary cytotoxic effects of SA-treated microtubers against the breast cancer cell line SUM159,comparable to field-grown tubers.Transcriptomic analysis revealed many differentially expressed genes(DEGs)in SA-treated in vitro microtubers.Six and twelve DEGs were annotated to the tropane,piperidine,and pyridine alkaloid pathway and the isoquinoline alkaloid pathway,respectively.RT-qPCR confirmed that the genes encoding spermidine synthase(c64642_g1),hyoscyamine(6S)-dioxygenase(c62620_g1),catechol oxidase(c61704_g3),monoamine oxidase(c65996_g3),and aspartate transaminase(c71069_g1)were significantly induced by SA.This study advances the production of P.ternata microtubers without field cultivation and provides considerable genetic information regarding SA-promoted alkaloid accumulation in P.ternata.展开更多
Hyperactivation of DNA repairing pathway is highly associated with the chemosensitivity and chemoresistance of cancer cells.In this manuscript,guided by cascaded one strain many compounds-global natural products socia...Hyperactivation of DNA repairing pathway is highly associated with the chemosensitivity and chemoresistance of cancer cells.In this manuscript,guided by cascaded one strain many compounds-global natural products social molecular networking(OSMAC-GNPS)strategy,a pair of epimeric environmental-induced metabolites were isolated from Aspergillus sp.EGF 15-0-3.Structurally,sterpiperazines A(1)and B(2)represent the first steroid-based indole alkaloids with unprecedented backbones.Biologically,compound 1 could be identified as a novel tyrosyl-DNA phosphodiesterase 1(Tdp1)inhibitor with a unique mechanism distinct from the reported modulators,and was able to significantly enhance the sensitivity of NCI-H460 cells to the clinic chemotherapeutic drug through inhibiting the DNA repairment and enhanced the DNA damage of cancer cells.展开更多
Coptis chinensis Franch.and Panax ginseng C.A.Mey.are traditional herbal medicines with millennia of documented use and broad therapeutic applications,including anti-diabetic properties.However,the synergistic effect ...Coptis chinensis Franch.and Panax ginseng C.A.Mey.are traditional herbal medicines with millennia of documented use and broad therapeutic applications,including anti-diabetic properties.However,the synergistic effect of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng on type 2 diabetes mellitus(T2DM)and its underlying mechanism remain unclear.The research demonstrated that the optimal ratio of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng was 4∶1,exhibiting maximal efficacy in improving insulin resistance and gluconeogenesis in primary mouse hepatocytes.This combination demonstrated significant synergistic effects in improving glucose tolerance,reducing fasting blood glucose(FBG),the weight ratio of epididymal white adipose tissue(eWAT),and the homeostasis model assessment of insulin resistance(HOMA-IR)in leptin receptor-deficient(db/db)mice.Subsequently,a T2DM liver-specific network was constructed based on RNA sequencing(RNA-seq)experiments and public databases by integrating transcriptional properties of disease-associated proteins and protein-protein interactions(PPIs).The network recovery index(NRI)score of the combined treatment group with a 4∶1 ratio exceeded that of groups treated with individual components.The research identified that activated adenosine 5'-monophosphate-activated protein kinase(AMPK)/acetyl-CoA carboxylase(ACC)signaling in the liver played a crucial role in the synergistic treatment of T2DM,as verified by western blot experiment in db/db mice.These findings demonstrate that the 4∶1 combination of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng significantly improves insulin resistance and glucose and lipid metabolism disorders in db/db mice,surpassing the efficacy of individual treatments.The synergistic mechanism correlates with enhanced AMPK/ACC signaling pathway activity.展开更多
Pyrrolizidine alkaloids(PAs)and their N-oxides(PANOs)are phytotoxins produced by various plant species and have been emerged as environmental pollutants.The sorption/desorption behaviors of PAs/PANOs in soil are cruci...Pyrrolizidine alkaloids(PAs)and their N-oxides(PANOs)are phytotoxins produced by various plant species and have been emerged as environmental pollutants.The sorption/desorption behaviors of PAs/PANOs in soil are crucial due to the horizontal transfer of these natural products from PA-producing plants to soil and subsequently absorbed by plant roots.This study firstly investigated the sorption/desorption behaviors of PAs/PANOs in tea plantation soils with distinct characteristics.Sorption amounts for seneciphylline(Sp)and seneciphylline-N-oxide(SpNO)in three acidic soils ranged from 2.9 to 5.9μg/g and 1.7 to 2.8μg/g,respectively.Desorption percentages for Sp and SpNO were from 22.2%to 30.5%and 36.1%to 43.9%.In the mixed PAs/PANOs systems,stronger sorption of PAs over PANOs was occurred in tested soils.Additionally,the Freundlich models more precisely described the sorption/desorption isotherms.Cation exchange capacity,sand content and total nitrogen were identified as major influencing factors by linear regression models.Overall,the soils exhibiting higher sorption capacities for compounds with greater hydrophobicity.PANOs were more likely to migrate within soils and be absorbed by tea plants.It contributes to the understanding of environmental fate of PAs/PANOs in tea plantations and provides basic data and clues for the development of PAs/PANOs reduction technology.展开更多
Aconitum(Ranunculaceae)has a long-standing history in traditional Chinese medicine(TCM),where it has been widely used to treat conditions such as rheumatoid arthritis(RA),myocardial infarction,and heart failure.Howeve...Aconitum(Ranunculaceae)has a long-standing history in traditional Chinese medicine(TCM),where it has been widely used to treat conditions such as rheumatoid arthritis(RA),myocardial infarction,and heart failure.However,the potency of Aconitum alkaloids,the primary active components of Aconitum,also confers substantial toxicity.Therefore,assessing the efficacy and toxicity of these Aconitum alkaloids is crucial for ensuring clinical effectiveness and safety.Metabolomics,a quantitative method for analyzing low-molecular-weight metabolites involved in metabolic pathways,provides a comprehensive view of the metabolic state across multiple systems in vivo.This approach has become a vital investigative tool for facilitating the evaluation of their efficacy and toxicity,identifying potential sensitive biomarkers,and offering a promising avenue for elucidating the pharmacological and toxicological mechanisms underlying TCM.This review focuses on the applications of metabolomics in pharmacological and toxicological studies of Aconitum alkaloids in recent years and highlights the significant role of metabolomics in exploring compatibility detoxification and the mechanisms of TCM processing,aiming to identify more viable methods for characterizing toxic medicinal plants.展开更多
From the seeds of Peganum harmala L.,three new alkaloids ofβ-carboline were isolated.Among them,peganumiums A(1)and B(2)were dimers with specific new scaffolds,all with long conjugated systems.Peganumium A and peganu...From the seeds of Peganum harmala L.,three new alkaloids ofβ-carboline were isolated.Among them,peganumiums A(1)and B(2)were dimers with specific new scaffolds,all with long conjugated systems.Peganumium A and peganumium C(3)were ionic alkaloid salts and peganumium B was a hexacycliccondensed alkaloid.The biosynthetic pathways of the three compounds above were also speculated.A preliminary cytotoxicity assay revealed that peganumium B had strong in vitro antiproliferative ability against a variety of cancer cells.The analysis of^(1)H nuclear magnetic resonance(NMR)metabolomics suggested that the antiproliferative mechanism of peganumium B could be associated with the biosynthesis of phenylalanine,tyrosine and tryptophan,the metabolism of glycine,serine,and threonine,the metabolism of taurine and hypotaurine,and the metabolism of nicotinate and nicotinamide.In addition,peganumium B could reduce the mitochondrial content of body-wall muscle cells of a Caenorhabditis elegans(C.elegans)strain in vivo.展开更多
Alduronic acid lactones and glyconolactones are highly functionalized and versatile chiral building blocks.Herein,we describe a novel approach to these compounds via decarboxylative oxygenation of uronic acids.The tra...Alduronic acid lactones and glyconolactones are highly functionalized and versatile chiral building blocks.Herein,we describe a novel approach to these compounds via decarboxylative oxygenation of uronic acids.The transformations proceed using Selectfluor and TEMPO as oxidants,either in the presence of catalytic amounts of Ag_(2)CO_(3)or in the absence of this catalyst.The methodology provides structurally diverse alduronic acid lactones and enables the preparation of rare sugar glyconolactones from easily available D-C-glycosides.Based on the^(18)O-labeling experiments,control experiments,and isolation of the key intermediates,a radical-polar crossover reaction mechanism is proposed.The utility of this method is demonstrated through efficient conversions of alduronic acid lactones into polyhydroxylated cyclic alkaloids and castanospermine-type architectures.展开更多
Inflammation plays a pivotal role in the etiology and progression of various diseases.In traditional Chinese medicine,the whole plants of Thesium chinense Turcz.and its preparations(e.g.Bairui Granules)have been emplo...Inflammation plays a pivotal role in the etiology and progression of various diseases.In traditional Chinese medicine,the whole plants of Thesium chinense Turcz.and its preparations(e.g.Bairui Granules)have been employed to manage inflammatory conditions.While flavonoids were previously considered the primary anti-inflammatory components,other potentially active constituents have been largely overlooked and not thoroughly investigated.This study presents a novel finding that the total alkaloids of T.chinense(BC-Alk)are potent active substances underlying the traditional and clinical applications of T.chinense and Bairui Granules as anti-inflammatory agents.UPLC-MS/MS analysis identified the composition of BC-Alk as quinolizidine alkaloids.The anti-inflammatory efficacy of BC-Alk was evaluated using a lipopolysaccharide(LPS)-induced lung inflammation model in mice.Results demonstrated that BC-Alk significantly mitigated LPS-induced lung inflammation,attenuated the overproduction of IL-1β and the overproduction of inflammatory factors(TNF-α),and ameliorated lung tissue hyperplasia in mice in vivo.Mechanistic studies in vitro revealed that BC-Alk upregulated the expression of Nrf2 and its downstream proteins NQO1 and glutamate-cystine ligase and modifier subunit(GCLM),inhibited NF-κB phosphorylation,and suppressed NLRP3 activation.Collectively,these findings indicate that BC-Alk exerts potent inhibitory effects against lung inflammation by modulating Nrf2,NF-κB,and NLRP3 pathways.This study provides new insights into the anti-inflammatory constituents of T.chinense and Bairui Granules.展开更多
The rapid screening of bioactive constituents within traditional Chinese medicine(TCM)presents a significant challenge to researchers.Prevailing strategies for the screening of active components in TCM often overlook ...The rapid screening of bioactive constituents within traditional Chinese medicine(TCM)presents a significant challenge to researchers.Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents.To address this limitation,a fishing strategy based on offline two-dimensional liquid chromatography(2D-LC)combined with surface plasmon resonance(SPR)was utilized to screen bioactive trace components targeting peroxiredoxin 3(PRDX3),using Uncaria alkaloids(UAs)as a case study.Initially,an orthogonal preparative offline 2D-LC system combining a positively charged C_(18)column and a conventional C_(18)column under disparate mobile phase conditions was constructed.To fully reveal the trace alkaloids,132D fractions of UAs were prepared,and their components were characterized using mass spectrometry(MS).Subsequently,employing PRDX3 as the targeting protein,a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether(GSM)serving as a positive control for binding.Employing this refined strategy,29 candidate binding alkaloids were fished from the 132D fractions.Notably,combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone.Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids,with isovallesiachotamine(IV),corynoxeine N-oxide(CO-N),and cadambine(CAD)demonstrating the highest affinity for PRDX3.Their interactions were further validated through molecular docking analysis.Subsequent intracellular H_(2)O_(2)measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H_(2)O_(2)clearance.In conclusion,this study introduced an innovative strategy for the identification of active trace components in TCM.This approach holds promise for accelerating research on medicinal components within this field.展开更多
Aconiti Lateralis Radix Praeparata(Fuzi)represents a significant traditional Chinese medicine(TCM)that exhibits both notable pharmacological effects and toxicity.Various processing methods are implemented to reduce th...Aconiti Lateralis Radix Praeparata(Fuzi)represents a significant traditional Chinese medicine(TCM)that exhibits both notable pharmacological effects and toxicity.Various processing methods are implemented to reduce the toxicity of raw Fuzi by modifying its toxic and effective components,primarily diterpenoid alkaloids.To comprehensively analyze the chemical variations between different Fuzi products,ultra-high performance liquid chromatography-linear ion trap quadrupole Orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS)was employed to systematically characterize Shengfuzi,Heishunpian and Baifupian.A total of 249 diterpenoid alkaloids present in Shengfuzi were identified,while only 111 and 61 in Heishunpian and Baifupian were detected respectively,indicating substantial differences among these products.An untargeted metabolomics approach combined with multivariate statistical analysis revealed 42 potential chemical markers.Through subsequent validation using 52 batches of commercial Heishunpian and Baifupian samples,8 robust markers distinguishing these products were identified,including AC1-propanoic acid-3OH,HE-glucoside,HE-hydroxyvaleric acid-2OH,dihydrosphingosine,N-dodecoxycarbonylvaline and three unknown compounds.Additionally,the MS imaging(MSI)technique was utilized to visualize the spatial distribution of chemical constituents in raw Fuzi,revealing how different processing procedures affect the chemical variations between Heishunpian and Baifupian.The distribution patterns of different diterpenoid alkaloid subtypes partially explained the chemical differences among products.This research provides valuable insights into the material basis for future investigations of different Fuzi products.展开更多
Peganum harmala L.(P.harmala)is a significant economic and medicinal plant.The seeds of P.harmala have been extensively utilized in traditional Chinese medicine,Uighur medicine,and Mongolian medicine,as documented in ...Peganum harmala L.(P.harmala)is a significant economic and medicinal plant.The seeds of P.harmala have been extensively utilized in traditional Chinese medicine,Uighur medicine,and Mongolian medicine,as documented in the Drug Standard of the Ministry of Health of China.Twelve novel tryptamine-derived alkaloids(1−12)and eight known compounds(13−20)were isolated from P.harmala seeds.Compounds 1 and 2 represent the first reported instances of tryptamine-derived heteromers,comprising tryptamine and aniline fragments with previously undocumented C-3−N-1′linkage and C-3−C-4′connection,respectively.Compounds 3−5 were identified as indole-quinazoline heteromers,exhibiting a novel C-3 and NH-1′linkage between indole and quinazoline-derived fragments.Compound 6 demonstrates the dimerization pattern of C-C linked tryptamine-quinazoline dimer.Compound 8 represents a tryptamine-derived heterodimer with a distinctive carbon skeleton,featuring an unusual spiro-tricyclic ring(7)and conventional bicyclic tryptamine.Compounds 9−11 constitute novel 6/5/5/5 spiro-tetracyclic tryptamine-derived alkaloids presenting a unique ring system of tryptamine-spiro-pyrrolizine.Compounds 1−3 and 6−11 were identified as racemates.Compounds 2,7,9,10,and 12 were confirmed via X-ray crystallographic analysis.All isolated compounds(1−20)exhibited varying degrees of antiviral efficacy against respiratory syncytial virus(RSV).Notably,the anti-RSV activity of compound 12(IC505.01±0.14μmol·L^(−1))surpassed that of the positive control(ribavirin,IC506.23±0.95μmol·L^(−1)),as validated through plaque reduction and immunofluorescence assays.The identification of anti-RSV compounds from P.harmala seeds may enhance the development and application of this plant in antiviral therapeutic products.展开更多
Anisodus tanguticus (Maxim.) Pascher, a medicinal plant in the Solanaceae family, is widely distributed across the Qinghai-Tibet Plateau. Its medicinal properties, particularly the tropane alkaloids, are influenced by...Anisodus tanguticus (Maxim.) Pascher, a medicinal plant in the Solanaceae family, is widely distributed across the Qinghai-Tibet Plateau. Its medicinal properties, particularly the tropane alkaloids, are influenced by various ecological factors, but the underlying ecological mechanisms remain poorly understood. This study aimed to investigate how key environmental variables influence both the morphological traits and tropane alkaloid contents of A. tanguticus, with the goal of providing data to support the sustainable cultivation and management of this species. We collected samples from 71 sites across its natural habitat, analyzing the effects of factors such as soil nutrients, altitude, and climate variables on plant traits and alkaloid composition. Statistical methods including Pearson correlation analysis, multiple regression, random forest analysis, and structural equation modeling were used to identify key environmental drivers. Our results indicate that available phosphorus significantly affects aboveground traits, while Cu concentration is most influential for root development. Altitude and longitude were found to be the main determinants of biomass accumulation. Regarding alkaloid content, Mg concentration in the soil was closely linked to anisodine levels, while altitude and latitude were the primary factors influencing anisodamine and atropine content, respectively. These findings provide essential insights into the ecological factors that govern the growth and medicinal compound production in A. tanguticus. Our research not only contributes to understanding the plant’s ecological requirements but also offers practical guidelines for selecting optimal cultivation conditions to enhance both yield and alkaloid quality, supporting sustainable use and conservation of this valuable medicinal resource.展开更多
ChuanWu(CW),the dried mother root of Aconitum carmichaelii Debx.,is a well-known traditional Chinese medicine(TCM)recognized for its potent efficacy but inherent toxicity,primarily due to its alkaloid content.Traditio...ChuanWu(CW),the dried mother root of Aconitum carmichaelii Debx.,is a well-known traditional Chinese medicine(TCM)recognized for its potent efficacy but inherent toxicity,primarily due to its alkaloid content.Traditional and modern detoxification methods for CW include proper processing,rational compatibility,and specialized decoction techniques,among which honey-boiled CW is particularly distinctive.However,research on the detoxification mechanism of honey-boiled CW remains limited.This study investigated this mechanism by analyzing alkaloid transformation and supramolecular aggregation.Honey-boiled and water-boiled CW preparations were compared.Ultra-high-performance liquid chromatography-tandem mass spectrometry was used to analyze CW alkaloids,specifically diester alkaloids(DDAs),monoester alkaloids(MDAs),and non-esterified diterpenoid alkaloids(NDAs).Transmission electron microscopy was employed to observe and identify supramolecular aggregates in the honey-boiled CW decoction.In vivo absorption of water-boiled,honey-boiled,and NADES-boiled CW was compared.Median lethal dose(LD_(50))tests assessed toxicity,including hepatotoxicity and nephrotoxicity.In vitro experiments evaluated the safety,anti-inflammatory,and analgesic effects of CWmedicated serum on RAW264.7 cells,with in vivo validation in mice.Results showed that honey promoted the conversion of highly toxic DDAs to less toxic MDAs and prevented MDAs from hydrolyzing into NDAs.Honey-boiled CW formed approximately 250 nm supramolecular aggregates that encapsulated MDAs,inhibiting their conversion to NDAs.These encapsulated MDAs acted as a stable delivery system with higher bioavailability than free benzoylmesaconine.Subsequent mouse experiments confirmed that honey-boiled CW significantly increased the LD_(50)of CW while reducing hepatotoxicity and nephrotoxicity.Additionally,honey-boiled CW significantly improved cell safety and enhanced anti-inflammatory and analgesic effects.Our findings reveal that honey-boiled CW exhibits a potent detoxification mechanism by influencing alkaloid transformation and facilitating the formation of supramolecular aggregates.This study lays the groundwork for developing detoxification or synergistic strategies within honey-boiled TCM.展开更多
Benzylisoquinoline alkaloids(BIAs)are a structurally diverse group of plant metabolites renowned for their pharmacological properties.However,sustainable sources for these compounds remain limited.Consequently,researc...Benzylisoquinoline alkaloids(BIAs)are a structurally diverse group of plant metabolites renowned for their pharmacological properties.However,sustainable sources for these compounds remain limited.Consequently,researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches.CYP80B,a family of cytochrome P450(CYP450)enzymes,plays a crucial role in BIA biosynthesis.Previously reported CYP80Bs are known to catalyze the 3′-hydroxylation of(S)-Nmethylcoclaurine,with the N-methyl group essential for catalytic activity.In this study,we successfully cloned a full-length CYP80B gene(St CYP80B)from Stephania tetrandra(S.tetrandra)and identified its function using a yeast heterologous expression system.Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that St CYP80B could catalyze Nmethylcoclaurine and coclaurine into their respective 3'-hydroxylated products.Notably,St CYP80B exhibited an expanded substrate selectivity compared to previously reported wildtype CYP80Bs,as it did not require an N-methyl group for hydroxylase activity.Furthermore,St CYP80B displayed a clear preference for the(S)-configuration.Co-expression of St CYP80B with the CYP450 reductases(CPRs,StCPR1,and StCPR2),also cloned from S.tetrandra,significantly enhanced the catalytic activity towards(S)-coclaurine.Site-directed mutagenesis of St CYP80B revealed that the residue H205 is crucial for coclaurine catalysis.Additionally,St CYP80B exhibited tissue-specific expression in plants.This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.展开更多
Background and objective:Both type 2 diabetes mellitus(T2DM)and metabolic dysfunction-associated steatotic liver disease(MASLD)are known to be influenced by environmental and lifestyle factors.Ramulus Mori(Sangzhi)alk...Background and objective:Both type 2 diabetes mellitus(T2DM)and metabolic dysfunction-associated steatotic liver disease(MASLD)are known to be influenced by environmental and lifestyle factors.Ramulus Mori(Sangzhi)alkaloids(SZ-A)are effective hypoglycemic agents.Recent studies suggest that SZ-A may improve T2DM,MASLD,and metabolic syndrome,but the underlying mechanisms remain unclear.This study aimed to investigate whether SZ-A can modulate hepatic lipid metabolism and gut microbiota in a mouse model of T2DM combined with MASLD.Methods:A combined T2DM-MASLD mouse model was established using a high-fat diet and streptozotocin injection.Liver morphology and histology were assessed using a portable small-animal ultrasound imaging system,hematoxylin and eosin(H&E)staining,and Oil Red O staining.Serum levels of triglycerides(TG),total cholesterol(TC),low-density lipoprotein(LDL),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)were measured using standard assay kits.Gut microbiota composition was analyzed by 16S rRNA sequencing,and hepatic lipid metabolites were profiled using liquid chromatography-mass spectrometry(LC-MS)MS.Results:SZ-A improved liver function by ameliorating morphological and structural abnormalities,reducing lipid droplet accumulation,and lowering serum levels of TG,TC,LDL,ALT,and AST.It also led to decreased hepatic ultrasound echo intensity compared to the kidney.Additionally,SZ-A helped restore gut microbiota balance,including a partial reversal of the Firmicutes/Bacteroidetes ratio.Lipidomic analysis revealed that SZ-A downregulated most TG and diglycerides(DG),while upregulating phosphatidylcholine(PC)and phosphatidylethanolamine(PE)in the model group.Conclusions:SZ-A partially alleviates liver injury in T2DM-MASLD mice by modulating hepatic lipid metabolism and gut microbiota composition.展开更多
The antiepileptic effect of pinellia total alkaloids（PTA） on penicillin（PNC） chronically kindled rats was investigated. We investigated the effects of PTA on Glu,Asp,Gly andγ-aminobutyric acid（GABA） concentrati...The antiepileptic effect of pinellia total alkaloids（PTA） on penicillin（PNC） chronically kindled rats was investigated. We investigated the effects of PTA on Glu,Asp,Gly andγ-aminobutyric acid（GABA） concentrations and the expression level of cerebral GABA_A receptor in hippocampus.The influence of PTA on epilepsy seizure latency and degree in PNC chronically kindled rats were observed.High performance liquid chromatography（HPLC） was adopted to measure the concentrations of Glu, Asp,Gly and GABA in hippocampus. Reverse transcriptase-polymerase chain reaction （RT-PCR） was used to determine the expression of cerebral GABAA receptor mRNA. Compared with normal rats, the levels of GABA and Gly decreased obviously while the level of Glu and Asp increased significantly in model rats. The cerebral GABAA receptor mRNA level was also decreased at the same time. The difference was statistically different compared to the control group. PTA could prolong the latent period of the penicillin induced seizure and weaken the extent of seizure, compared with the model group without PTA treatment. Moreover, PTA increased the level of GABA and the expression level of GABAA receptor, while decreased the level of Glu significantly. However, it had no obvious effect on the level of Gly and Asp. Pre-treatment of PTA can also increase the GABAA receptor mRNA level. In conclusion, PTA could alleviate the PNC chronically kindled rat seizure. It increased the GABA level and the expression of GABAA receptor, and it decreased the Glu concentration.展开更多
Two novel hydroperoxylated Lycopodium alkaloids, 11alpha-hydroperoxyphlegmariurine B (1) and 7-hydroperoxyphlegmariurine B (2), along with a known compound, phlegmanurine B (3), were isolated from the total alkaloid f...Two novel hydroperoxylated Lycopodium alkaloids, 11alpha-hydroperoxyphlegmariurine B (1) and 7-hydroperoxyphlegmariurine B (2), along with a known compound, phlegmanurine B (3), were isolated from the total alkaloid fraction of the Chinese medicinal herb Huperzia serrata (Thunb.) Trev. Their structures and relative configurations were elucidated on the basis of spectroscopic analyses.展开更多
Three new napelline-type C20-diterpenoid alkaloids,named aconicarmichinium A and B trifluoroacetates（1 and 2） and aconicarmichinium C chloride（3）,were isolated from an aqueous extract of ＂fu zi＂,the lateral root...Three new napelline-type C20-diterpenoid alkaloids,named aconicarmichinium A and B trifluoroacetates（1 and 2） and aconicarmichinium C chloride（3）,were isolated from an aqueous extract of ＂fu zi＂,the lateral roots of Aconitum carmichaelii.Their structures were elucidated by extensive spectroscopic data analysis.Compounds 1-3 represent the first examples of napelline-type C20 diterpenoid alkaloid alcohol iminiums,of which the structures were fully characterized.In addition,transformation and equilibration between the alcohol iminiums（1-3） and the aza acetals la-3a were investigated by measurements of the NMR spectra in protic and aprotic deuterium solvents including alkali pyridine-d5,along with evaporation under reduced pressure and gradual additions of TFA,AcOH,and HC1.The results demonstrated that the transformation and equilibration were solvent-,base-,and acid-dependent.Especially,in aqueous biological fluid,these C20-diterpenoid alkaloids would more likely exist as the alcohol iminiums accompanied by anion counterparts in biosystems to increase their solubility, bioavailability, transportations, and functions.The absolute configurations of 1-3 were confirmed by X-ray crystallographic analysis of 2a.展开更多
Two new norditerpenoid alkaloids, orthocentrine (1) and deacetylswinanine A (2), together with a known alkaloid, swinanine A (3), were isolated from the whole plants of Delphinium orthocentrum Franch. The structures o...Two new norditerpenoid alkaloids, orthocentrine (1) and deacetylswinanine A (2), together with a known alkaloid, swinanine A (3), were isolated from the whole plants of Delphinium orthocentrum Franch. The structures of the new alkaloids (1 and 2) were elucidated as 7,10_dihydroxy_8,14,16_trimethoxy_19,20_didehydro_aconitane (7β,8β,14α,16β) (1) and 20_ethyl_2,3_didehydro_6,10_dihydroxy_7,8_methylenedioxy_1,14,16_trimethoxy_aconitane (1α,6β,14α,16β) (2) by spectroscopic evidence and chemical transformation.展开更多
A reversed-phase high-performance liquid chromatography （RP-HPLC） method with diode array detector was established to simultaneously determine nine bioactive compounds in Wuzhuyu decoction, namely dehydroevodiamine,...A reversed-phase high-performance liquid chromatography （RP-HPLC） method with diode array detector was established to simultaneously determine nine bioactive compounds in Wuzhuyu decoction, namely dehydroevodiamine, isorhanmetin-7-O- rutinoside, 10-hydroxyrutaecarpine, diosmetin-7-O-13-D-glucopyranoside, evodiamine, rutaecarpine, 1-metbyl-2-n-nonyl-4（1H） quinolone, evocarpine, and dihydroevocarpine. The RP-HPLC assay was performed on a reversed-phase Cl8 column with a gradient elution. The mobile phases consisted of methanol and water containing 1% （v/v） acetic acid. The flow rate was 1.0 mL/min and the detection wavelength was set at 300 nm. All calibration curves showed good linearity （r2〉0.9941） within test ranges. The results of intra-day precision, inter-day precision, and accuracy were satisfactory. The overall recoveries were in the range of 90.13%-102.48%. The method was successfully applied to quantify nine bioactive compounds in six samples. Baseline resolution for all nine major compounds was achieved. This method could be used as a suitable quality control assay for Wuzhuyu decoction.展开更多
基金supported by the Key Project of Natural Science Research for Colleges and Universities in Anhui Province(2024AH051661,2023AH050345)Excellent Scientific Research and Innovation Team of University in Anhui Province(2022AH010029).
文摘As its tuberous alkaloids induce valuable pharmacological effects,Pinellia ternata has considerable clinical value.However,its production currently fails to meet its demand.In vitro microtuber culture,combined with salicylic acid(SA)elicitation,may provide an effective alternative to traditional field production.In this study,an in vitro P.ternata microtuber induction system was developed and used to evaluate SA-induced elicitation of alkaloid accumulation.The quality of in vitro microtubers was assessed by total alkaloid measurement,a cytotoxicity assay,and transcriptomic analysis.With or without SA treatment,P.ternata microtuber induction was achieved within 60 d using petiole-derived explants,with a microtuber proliferation rate of approximately 17 microtubers per explant.The total alkaloid content of in vitro microtubers elicited with 100μM SA was equivalent to that of field-grown tubers,while those not treated with SA contained lower alkaloid content.The cytotoxicity assay showed preliminary cytotoxic effects of SA-treated microtubers against the breast cancer cell line SUM159,comparable to field-grown tubers.Transcriptomic analysis revealed many differentially expressed genes(DEGs)in SA-treated in vitro microtubers.Six and twelve DEGs were annotated to the tropane,piperidine,and pyridine alkaloid pathway and the isoquinoline alkaloid pathway,respectively.RT-qPCR confirmed that the genes encoding spermidine synthase(c64642_g1),hyoscyamine(6S)-dioxygenase(c62620_g1),catechol oxidase(c61704_g3),monoamine oxidase(c65996_g3),and aspartate transaminase(c71069_g1)were significantly induced by SA.This study advances the production of P.ternata microtubers without field cultivation and provides considerable genetic information regarding SA-promoted alkaloid accumulation in P.ternata.
基金supported by the National Natural Science Foundation of China(Nos.82273845,82304331,and 82360695)Guangxi Natural Science Foundation project(No.2023GXNSFBA026305).
文摘Hyperactivation of DNA repairing pathway is highly associated with the chemosensitivity and chemoresistance of cancer cells.In this manuscript,guided by cascaded one strain many compounds-global natural products social molecular networking(OSMAC-GNPS)strategy,a pair of epimeric environmental-induced metabolites were isolated from Aspergillus sp.EGF 15-0-3.Structurally,sterpiperazines A(1)and B(2)represent the first steroid-based indole alkaloids with unprecedented backbones.Biologically,compound 1 could be identified as a novel tyrosyl-DNA phosphodiesterase 1(Tdp1)inhibitor with a unique mechanism distinct from the reported modulators,and was able to significantly enhance the sensitivity of NCI-H460 cells to the clinic chemotherapeutic drug through inhibiting the DNA repairment and enhanced the DNA damage of cancer cells.
基金supported by the Pioneer and Leading Goose R&D Program of Zhejiang Province(No.2024C03106)the National Natural Science Foundation of China(No.U23A20513)+1 种基金Ningbo Top Medical and Health Research Program(No.2022030309)the Innovation Team and Talents Cultivation Program of the National Administration of Traditional Chinese Medicine(No.ZYYCXTD-D-202002).
文摘Coptis chinensis Franch.and Panax ginseng C.A.Mey.are traditional herbal medicines with millennia of documented use and broad therapeutic applications,including anti-diabetic properties.However,the synergistic effect of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng on type 2 diabetes mellitus(T2DM)and its underlying mechanism remain unclear.The research demonstrated that the optimal ratio of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng was 4∶1,exhibiting maximal efficacy in improving insulin resistance and gluconeogenesis in primary mouse hepatocytes.This combination demonstrated significant synergistic effects in improving glucose tolerance,reducing fasting blood glucose(FBG),the weight ratio of epididymal white adipose tissue(eWAT),and the homeostasis model assessment of insulin resistance(HOMA-IR)in leptin receptor-deficient(db/db)mice.Subsequently,a T2DM liver-specific network was constructed based on RNA sequencing(RNA-seq)experiments and public databases by integrating transcriptional properties of disease-associated proteins and protein-protein interactions(PPIs).The network recovery index(NRI)score of the combined treatment group with a 4∶1 ratio exceeded that of groups treated with individual components.The research identified that activated adenosine 5'-monophosphate-activated protein kinase(AMPK)/acetyl-CoA carboxylase(ACC)signaling in the liver played a crucial role in the synergistic treatment of T2DM,as verified by western blot experiment in db/db mice.These findings demonstrate that the 4∶1 combination of total alkaloids from Coptis chinensis and total ginsenosides from Panax ginseng significantly improves insulin resistance and glucose and lipid metabolism disorders in db/db mice,surpassing the efficacy of individual treatments.The synergistic mechanism correlates with enhanced AMPK/ACC signaling pathway activity.
基金supported by the earmarked fund for the Modern Agro-Industry Technology Research System (No.CARS-19)the Innovative Research Team in Chinese Academy of Agricultural Sciences (No.CAAS ASTIP-2014-TRICAAS).
文摘Pyrrolizidine alkaloids(PAs)and their N-oxides(PANOs)are phytotoxins produced by various plant species and have been emerged as environmental pollutants.The sorption/desorption behaviors of PAs/PANOs in soil are crucial due to the horizontal transfer of these natural products from PA-producing plants to soil and subsequently absorbed by plant roots.This study firstly investigated the sorption/desorption behaviors of PAs/PANOs in tea plantation soils with distinct characteristics.Sorption amounts for seneciphylline(Sp)and seneciphylline-N-oxide(SpNO)in three acidic soils ranged from 2.9 to 5.9μg/g and 1.7 to 2.8μg/g,respectively.Desorption percentages for Sp and SpNO were from 22.2%to 30.5%and 36.1%to 43.9%.In the mixed PAs/PANOs systems,stronger sorption of PAs over PANOs was occurred in tested soils.Additionally,the Freundlich models more precisely described the sorption/desorption isotherms.Cation exchange capacity,sand content and total nitrogen were identified as major influencing factors by linear regression models.Overall,the soils exhibiting higher sorption capacities for compounds with greater hydrophobicity.PANOs were more likely to migrate within soils and be absorbed by tea plants.It contributes to the understanding of environmental fate of PAs/PANOs in tea plantations and provides basic data and clues for the development of PAs/PANOs reduction technology.
基金supported by the National Natural Science Foundation of China (No.82274223)。
文摘Aconitum(Ranunculaceae)has a long-standing history in traditional Chinese medicine(TCM),where it has been widely used to treat conditions such as rheumatoid arthritis(RA),myocardial infarction,and heart failure.However,the potency of Aconitum alkaloids,the primary active components of Aconitum,also confers substantial toxicity.Therefore,assessing the efficacy and toxicity of these Aconitum alkaloids is crucial for ensuring clinical effectiveness and safety.Metabolomics,a quantitative method for analyzing low-molecular-weight metabolites involved in metabolic pathways,provides a comprehensive view of the metabolic state across multiple systems in vivo.This approach has become a vital investigative tool for facilitating the evaluation of their efficacy and toxicity,identifying potential sensitive biomarkers,and offering a promising avenue for elucidating the pharmacological and toxicological mechanisms underlying TCM.This review focuses on the applications of metabolomics in pharmacological and toxicological studies of Aconitum alkaloids in recent years and highlights the significant role of metabolomics in exploring compatibility detoxification and the mechanisms of TCM processing,aiming to identify more viable methods for characterizing toxic medicinal plants.
基金Financial support from Beijing Natural Science Foundation(No.7232283)。
文摘From the seeds of Peganum harmala L.,three new alkaloids ofβ-carboline were isolated.Among them,peganumiums A(1)and B(2)were dimers with specific new scaffolds,all with long conjugated systems.Peganumium A and peganumium C(3)were ionic alkaloid salts and peganumium B was a hexacycliccondensed alkaloid.The biosynthetic pathways of the three compounds above were also speculated.A preliminary cytotoxicity assay revealed that peganumium B had strong in vitro antiproliferative ability against a variety of cancer cells.The analysis of^(1)H nuclear magnetic resonance(NMR)metabolomics suggested that the antiproliferative mechanism of peganumium B could be associated with the biosynthesis of phenylalanine,tyrosine and tryptophan,the metabolism of glycine,serine,and threonine,the metabolism of taurine and hypotaurine,and the metabolism of nicotinate and nicotinamide.In addition,peganumium B could reduce the mitochondrial content of body-wall muscle cells of a Caenorhabditis elegans(C.elegans)strain in vivo.
基金financial support from the Marine S&T Fund of Shandong Province for Qingdao Marine Science and Technology Center(No.2022QNLM030003-2)the National Key Research&Development Program of China(No.2022YFA2104902)the National Natural Science Foundation of China(Nos.21977088 and22377114)。
文摘Alduronic acid lactones and glyconolactones are highly functionalized and versatile chiral building blocks.Herein,we describe a novel approach to these compounds via decarboxylative oxygenation of uronic acids.The transformations proceed using Selectfluor and TEMPO as oxidants,either in the presence of catalytic amounts of Ag_(2)CO_(3)or in the absence of this catalyst.The methodology provides structurally diverse alduronic acid lactones and enables the preparation of rare sugar glyconolactones from easily available D-C-glycosides.Based on the^(18)O-labeling experiments,control experiments,and isolation of the key intermediates,a radical-polar crossover reaction mechanism is proposed.The utility of this method is demonstrated through efficient conversions of alduronic acid lactones into polyhydroxylated cyclic alkaloids and castanospermine-type architectures.
基金supported by the National Natural Science Foundation of China(No.82274065)Toxicology of Traditional Chinese Medicines,a High-Level Priority Subject in Chinese Medicine in National Administration of Traditional Chinese Medicine(No.ZYYZDXK-2023296)Cutting Edge Development Fund of Advanced Medical Research Institute(No.GYY2023QY01)。
文摘Inflammation plays a pivotal role in the etiology and progression of various diseases.In traditional Chinese medicine,the whole plants of Thesium chinense Turcz.and its preparations(e.g.Bairui Granules)have been employed to manage inflammatory conditions.While flavonoids were previously considered the primary anti-inflammatory components,other potentially active constituents have been largely overlooked and not thoroughly investigated.This study presents a novel finding that the total alkaloids of T.chinense(BC-Alk)are potent active substances underlying the traditional and clinical applications of T.chinense and Bairui Granules as anti-inflammatory agents.UPLC-MS/MS analysis identified the composition of BC-Alk as quinolizidine alkaloids.The anti-inflammatory efficacy of BC-Alk was evaluated using a lipopolysaccharide(LPS)-induced lung inflammation model in mice.Results demonstrated that BC-Alk significantly mitigated LPS-induced lung inflammation,attenuated the overproduction of IL-1β and the overproduction of inflammatory factors(TNF-α),and ameliorated lung tissue hyperplasia in mice in vivo.Mechanistic studies in vitro revealed that BC-Alk upregulated the expression of Nrf2 and its downstream proteins NQO1 and glutamate-cystine ligase and modifier subunit(GCLM),inhibited NF-κB phosphorylation,and suppressed NLRP3 activation.Collectively,these findings indicate that BC-Alk exerts potent inhibitory effects against lung inflammation by modulating Nrf2,NF-κB,and NLRP3 pathways.This study provides new insights into the anti-inflammatory constituents of T.chinense and Bairui Granules.
基金supported by Shanghai Institute of Materia Medica-Shanghai University of Traditional Chinese Medicine(SIMM-SHUTCM)Traditional Chinese Medicine Innovation Joint Research Program,China(Grant No.:2022,E2G808H096)the National Key Research and Development Program of China(Grant Nos.:2022YFC3501704 and 2023YFC3504205)Sanming Project of Medicine in Shenzhen,China(Grant No.:ZZYSM202106004).
文摘The rapid screening of bioactive constituents within traditional Chinese medicine(TCM)presents a significant challenge to researchers.Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents.To address this limitation,a fishing strategy based on offline two-dimensional liquid chromatography(2D-LC)combined with surface plasmon resonance(SPR)was utilized to screen bioactive trace components targeting peroxiredoxin 3(PRDX3),using Uncaria alkaloids(UAs)as a case study.Initially,an orthogonal preparative offline 2D-LC system combining a positively charged C_(18)column and a conventional C_(18)column under disparate mobile phase conditions was constructed.To fully reveal the trace alkaloids,132D fractions of UAs were prepared,and their components were characterized using mass spectrometry(MS).Subsequently,employing PRDX3 as the targeting protein,a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether(GSM)serving as a positive control for binding.Employing this refined strategy,29 candidate binding alkaloids were fished from the 132D fractions.Notably,combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone.Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids,with isovallesiachotamine(IV),corynoxeine N-oxide(CO-N),and cadambine(CAD)demonstrating the highest affinity for PRDX3.Their interactions were further validated through molecular docking analysis.Subsequent intracellular H_(2)O_(2)measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H_(2)O_(2)clearance.In conclusion,this study introduced an innovative strategy for the identification of active trace components in TCM.This approach holds promise for accelerating research on medicinal components within this field.
基金supported by the Qi-Huang Chief Scientist Program of the National Administration of Traditional Chinese Medicine(2020)the National Key Research and Development Program of China(No.2022YFC3501705)+1 种基金Shanghai Sailing Program(No.23YF1447500)the China Postdoctoral Science Foundation(No.2023M732335).
文摘Aconiti Lateralis Radix Praeparata(Fuzi)represents a significant traditional Chinese medicine(TCM)that exhibits both notable pharmacological effects and toxicity.Various processing methods are implemented to reduce the toxicity of raw Fuzi by modifying its toxic and effective components,primarily diterpenoid alkaloids.To comprehensively analyze the chemical variations between different Fuzi products,ultra-high performance liquid chromatography-linear ion trap quadrupole Orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS)was employed to systematically characterize Shengfuzi,Heishunpian and Baifupian.A total of 249 diterpenoid alkaloids present in Shengfuzi were identified,while only 111 and 61 in Heishunpian and Baifupian were detected respectively,indicating substantial differences among these products.An untargeted metabolomics approach combined with multivariate statistical analysis revealed 42 potential chemical markers.Through subsequent validation using 52 batches of commercial Heishunpian and Baifupian samples,8 robust markers distinguishing these products were identified,including AC1-propanoic acid-3OH,HE-glucoside,HE-hydroxyvaleric acid-2OH,dihydrosphingosine,N-dodecoxycarbonylvaline and three unknown compounds.Additionally,the MS imaging(MSI)technique was utilized to visualize the spatial distribution of chemical constituents in raw Fuzi,revealing how different processing procedures affect the chemical variations between Heishunpian and Baifupian.The distribution patterns of different diterpenoid alkaloid subtypes partially explained the chemical differences among products.This research provides valuable insights into the material basis for future investigations of different Fuzi products.
基金supported by the National Natural Science Foundation of China(Nos.82104015,82003609,and 81973190)the Guangdong Basic and Applied Basic Research Foundation(Nos.2020A1515110415,2020A1515110453,and 2020B1515020033)+1 种基金the Characteristic Innovation Project of General Universities in Guangdong Province(No.2024KTSCX167)the Science and Technology Planning Project of Guangzhou City(No.202201010485).
文摘Peganum harmala L.(P.harmala)is a significant economic and medicinal plant.The seeds of P.harmala have been extensively utilized in traditional Chinese medicine,Uighur medicine,and Mongolian medicine,as documented in the Drug Standard of the Ministry of Health of China.Twelve novel tryptamine-derived alkaloids(1−12)and eight known compounds(13−20)were isolated from P.harmala seeds.Compounds 1 and 2 represent the first reported instances of tryptamine-derived heteromers,comprising tryptamine and aniline fragments with previously undocumented C-3−N-1′linkage and C-3−C-4′connection,respectively.Compounds 3−5 were identified as indole-quinazoline heteromers,exhibiting a novel C-3 and NH-1′linkage between indole and quinazoline-derived fragments.Compound 6 demonstrates the dimerization pattern of C-C linked tryptamine-quinazoline dimer.Compound 8 represents a tryptamine-derived heterodimer with a distinctive carbon skeleton,featuring an unusual spiro-tricyclic ring(7)and conventional bicyclic tryptamine.Compounds 9−11 constitute novel 6/5/5/5 spiro-tetracyclic tryptamine-derived alkaloids presenting a unique ring system of tryptamine-spiro-pyrrolizine.Compounds 1−3 and 6−11 were identified as racemates.Compounds 2,7,9,10,and 12 were confirmed via X-ray crystallographic analysis.All isolated compounds(1−20)exhibited varying degrees of antiviral efficacy against respiratory syncytial virus(RSV).Notably,the anti-RSV activity of compound 12(IC505.01±0.14μmol·L^(−1))surpassed that of the positive control(ribavirin,IC506.23±0.95μmol·L^(−1)),as validated through plaque reduction and immunofluorescence assays.The identification of anti-RSV compounds from P.harmala seeds may enhance the development and application of this plant in antiviral therapeutic products.
基金supported by Qinghai Science and Technology Achievement Transformation Project(2021-SF-149)Natural Science Fund of Education Department of Anhui Province(2024AH051669)+2 种基金Innovation and Entrepreneurship Training Program for College Students(S202410373040)Anhui Provincial College Student Innovation and Entrepreneurship Project(20221502032)School-Level Quality Engineering Project of Huaibei Normal University(2024xjsxyj003).
文摘Anisodus tanguticus (Maxim.) Pascher, a medicinal plant in the Solanaceae family, is widely distributed across the Qinghai-Tibet Plateau. Its medicinal properties, particularly the tropane alkaloids, are influenced by various ecological factors, but the underlying ecological mechanisms remain poorly understood. This study aimed to investigate how key environmental variables influence both the morphological traits and tropane alkaloid contents of A. tanguticus, with the goal of providing data to support the sustainable cultivation and management of this species. We collected samples from 71 sites across its natural habitat, analyzing the effects of factors such as soil nutrients, altitude, and climate variables on plant traits and alkaloid composition. Statistical methods including Pearson correlation analysis, multiple regression, random forest analysis, and structural equation modeling were used to identify key environmental drivers. Our results indicate that available phosphorus significantly affects aboveground traits, while Cu concentration is most influential for root development. Altitude and longitude were found to be the main determinants of biomass accumulation. Regarding alkaloid content, Mg concentration in the soil was closely linked to anisodine levels, while altitude and latitude were the primary factors influencing anisodamine and atropine content, respectively. These findings provide essential insights into the ecological factors that govern the growth and medicinal compound production in A. tanguticus. Our research not only contributes to understanding the plant’s ecological requirements but also offers practical guidelines for selecting optimal cultivation conditions to enhance both yield and alkaloid quality, supporting sustainable use and conservation of this valuable medicinal resource.
基金supported by National Natural Science Foundation of China(Grant Nos.:82230117,82474195,and 82173992)Youth Medical Innovation Research Project of China(Grant No.:P24021887623)+1 种基金Nanjing Medical University(Grant Nos.:TZKY20230104,and 2024KF0292)Science and Technology Support Projectof Taizhou(Grant No.:TS202420).
文摘ChuanWu(CW),the dried mother root of Aconitum carmichaelii Debx.,is a well-known traditional Chinese medicine(TCM)recognized for its potent efficacy but inherent toxicity,primarily due to its alkaloid content.Traditional and modern detoxification methods for CW include proper processing,rational compatibility,and specialized decoction techniques,among which honey-boiled CW is particularly distinctive.However,research on the detoxification mechanism of honey-boiled CW remains limited.This study investigated this mechanism by analyzing alkaloid transformation and supramolecular aggregation.Honey-boiled and water-boiled CW preparations were compared.Ultra-high-performance liquid chromatography-tandem mass spectrometry was used to analyze CW alkaloids,specifically diester alkaloids(DDAs),monoester alkaloids(MDAs),and non-esterified diterpenoid alkaloids(NDAs).Transmission electron microscopy was employed to observe and identify supramolecular aggregates in the honey-boiled CW decoction.In vivo absorption of water-boiled,honey-boiled,and NADES-boiled CW was compared.Median lethal dose(LD_(50))tests assessed toxicity,including hepatotoxicity and nephrotoxicity.In vitro experiments evaluated the safety,anti-inflammatory,and analgesic effects of CWmedicated serum on RAW264.7 cells,with in vivo validation in mice.Results showed that honey promoted the conversion of highly toxic DDAs to less toxic MDAs and prevented MDAs from hydrolyzing into NDAs.Honey-boiled CW formed approximately 250 nm supramolecular aggregates that encapsulated MDAs,inhibiting their conversion to NDAs.These encapsulated MDAs acted as a stable delivery system with higher bioavailability than free benzoylmesaconine.Subsequent mouse experiments confirmed that honey-boiled CW significantly increased the LD_(50)of CW while reducing hepatotoxicity and nephrotoxicity.Additionally,honey-boiled CW significantly improved cell safety and enhanced anti-inflammatory and analgesic effects.Our findings reveal that honey-boiled CW exhibits a potent detoxification mechanism by influencing alkaloid transformation and facilitating the formation of supramolecular aggregates.This study lays the groundwork for developing detoxification or synergistic strategies within honey-boiled TCM.
文摘Benzylisoquinoline alkaloids(BIAs)are a structurally diverse group of plant metabolites renowned for their pharmacological properties.However,sustainable sources for these compounds remain limited.Consequently,researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches.CYP80B,a family of cytochrome P450(CYP450)enzymes,plays a crucial role in BIA biosynthesis.Previously reported CYP80Bs are known to catalyze the 3′-hydroxylation of(S)-Nmethylcoclaurine,with the N-methyl group essential for catalytic activity.In this study,we successfully cloned a full-length CYP80B gene(St CYP80B)from Stephania tetrandra(S.tetrandra)and identified its function using a yeast heterologous expression system.Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that St CYP80B could catalyze Nmethylcoclaurine and coclaurine into their respective 3'-hydroxylated products.Notably,St CYP80B exhibited an expanded substrate selectivity compared to previously reported wildtype CYP80Bs,as it did not require an N-methyl group for hydroxylase activity.Furthermore,St CYP80B displayed a clear preference for the(S)-configuration.Co-expression of St CYP80B with the CYP450 reductases(CPRs,StCPR1,and StCPR2),also cloned from S.tetrandra,significantly enhanced the catalytic activity towards(S)-coclaurine.Site-directed mutagenesis of St CYP80B revealed that the residue H205 is crucial for coclaurine catalysis.Additionally,St CYP80B exhibited tissue-specific expression in plants.This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.
基金supported by the Fund for Independent Innovation of Hypoglycemic Drugs from Basic Research to Clinical Application(070500020373).
文摘Background and objective:Both type 2 diabetes mellitus(T2DM)and metabolic dysfunction-associated steatotic liver disease(MASLD)are known to be influenced by environmental and lifestyle factors.Ramulus Mori(Sangzhi)alkaloids(SZ-A)are effective hypoglycemic agents.Recent studies suggest that SZ-A may improve T2DM,MASLD,and metabolic syndrome,but the underlying mechanisms remain unclear.This study aimed to investigate whether SZ-A can modulate hepatic lipid metabolism and gut microbiota in a mouse model of T2DM combined with MASLD.Methods:A combined T2DM-MASLD mouse model was established using a high-fat diet and streptozotocin injection.Liver morphology and histology were assessed using a portable small-animal ultrasound imaging system,hematoxylin and eosin(H&E)staining,and Oil Red O staining.Serum levels of triglycerides(TG),total cholesterol(TC),low-density lipoprotein(LDL),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)were measured using standard assay kits.Gut microbiota composition was analyzed by 16S rRNA sequencing,and hepatic lipid metabolites were profiled using liquid chromatography-mass spectrometry(LC-MS)MS.Results:SZ-A improved liver function by ameliorating morphological and structural abnormalities,reducing lipid droplet accumulation,and lowering serum levels of TG,TC,LDL,ALT,and AST.It also led to decreased hepatic ultrasound echo intensity compared to the kidney.Additionally,SZ-A helped restore gut microbiota balance,including a partial reversal of the Firmicutes/Bacteroidetes ratio.Lipidomic analysis revealed that SZ-A downregulated most TG and diglycerides(DG),while upregulating phosphatidylcholine(PC)and phosphatidylethanolamine(PE)in the model group.Conclusions:SZ-A partially alleviates liver injury in T2DM-MASLD mice by modulating hepatic lipid metabolism and gut microbiota composition.
基金Natural Science Foundation of Shanxi Province (Grant No.20041109).
文摘The antiepileptic effect of pinellia total alkaloids（PTA） on penicillin（PNC） chronically kindled rats was investigated. We investigated the effects of PTA on Glu,Asp,Gly andγ-aminobutyric acid（GABA） concentrations and the expression level of cerebral GABA_A receptor in hippocampus.The influence of PTA on epilepsy seizure latency and degree in PNC chronically kindled rats were observed.High performance liquid chromatography（HPLC） was adopted to measure the concentrations of Glu, Asp,Gly and GABA in hippocampus. Reverse transcriptase-polymerase chain reaction （RT-PCR） was used to determine the expression of cerebral GABAA receptor mRNA. Compared with normal rats, the levels of GABA and Gly decreased obviously while the level of Glu and Asp increased significantly in model rats. The cerebral GABAA receptor mRNA level was also decreased at the same time. The difference was statistically different compared to the control group. PTA could prolong the latent period of the penicillin induced seizure and weaken the extent of seizure, compared with the model group without PTA treatment. Moreover, PTA increased the level of GABA and the expression level of GABAA receptor, while decreased the level of Glu significantly. However, it had no obvious effect on the level of Gly and Asp. Pre-treatment of PTA can also increase the GABAA receptor mRNA level. In conclusion, PTA could alleviate the PNC chronically kindled rat seizure. It increased the GABA level and the expression of GABAA receptor, and it decreased the Glu concentration.
文摘Two novel hydroperoxylated Lycopodium alkaloids, 11alpha-hydroperoxyphlegmariurine B (1) and 7-hydroperoxyphlegmariurine B (2), along with a known compound, phlegmanurine B (3), were isolated from the total alkaloid fraction of the Chinese medicinal herb Huperzia serrata (Thunb.) Trev. Their structures and relative configurations were elucidated on the basis of spectroscopic analyses.
基金Financial support from the National Natural Science Foundation of China (NNSFC Nos. 21132009, 30825044)the National Science and Technology Project of China (Nos. 2012ZX09301002002, 2011ZX0 9307-002-01)
文摘Three new napelline-type C20-diterpenoid alkaloids,named aconicarmichinium A and B trifluoroacetates（1 and 2） and aconicarmichinium C chloride（3）,were isolated from an aqueous extract of ＂fu zi＂,the lateral roots of Aconitum carmichaelii.Their structures were elucidated by extensive spectroscopic data analysis.Compounds 1-3 represent the first examples of napelline-type C20 diterpenoid alkaloid alcohol iminiums,of which the structures were fully characterized.In addition,transformation and equilibration between the alcohol iminiums（1-3） and the aza acetals la-3a were investigated by measurements of the NMR spectra in protic and aprotic deuterium solvents including alkali pyridine-d5,along with evaporation under reduced pressure and gradual additions of TFA,AcOH,and HC1.The results demonstrated that the transformation and equilibration were solvent-,base-,and acid-dependent.Especially,in aqueous biological fluid,these C20-diterpenoid alkaloids would more likely exist as the alcohol iminiums accompanied by anion counterparts in biosystems to increase their solubility, bioavailability, transportations, and functions.The absolute configurations of 1-3 were confirmed by X-ray crystallographic analysis of 2a.
文摘Two new norditerpenoid alkaloids, orthocentrine (1) and deacetylswinanine A (2), together with a known alkaloid, swinanine A (3), were isolated from the whole plants of Delphinium orthocentrum Franch. The structures of the new alkaloids (1 and 2) were elucidated as 7,10_dihydroxy_8,14,16_trimethoxy_19,20_didehydro_aconitane (7β,8β,14α,16β) (1) and 20_ethyl_2,3_didehydro_6,10_dihydroxy_7,8_methylenedioxy_1,14,16_trimethoxy_aconitane (1α,6β,14α,16β) (2) by spectroscopic evidence and chemical transformation.
基金National Sciences and Technology Program of China (Grant No. 2011BAI07B08,2006BAI06A01-02,2006BAI08B03-09)
文摘A reversed-phase high-performance liquid chromatography （RP-HPLC） method with diode array detector was established to simultaneously determine nine bioactive compounds in Wuzhuyu decoction, namely dehydroevodiamine, isorhanmetin-7-O- rutinoside, 10-hydroxyrutaecarpine, diosmetin-7-O-13-D-glucopyranoside, evodiamine, rutaecarpine, 1-metbyl-2-n-nonyl-4（1H） quinolone, evocarpine, and dihydroevocarpine. The RP-HPLC assay was performed on a reversed-phase Cl8 column with a gradient elution. The mobile phases consisted of methanol and water containing 1% （v/v） acetic acid. The flow rate was 1.0 mL/min and the detection wavelength was set at 300 nm. All calibration curves showed good linearity （r2〉0.9941） within test ranges. The results of intra-day precision, inter-day precision, and accuracy were satisfactory. The overall recoveries were in the range of 90.13%-102.48%. The method was successfully applied to quantify nine bioactive compounds in six samples. Baseline resolution for all nine major compounds was achieved. This method could be used as a suitable quality control assay for Wuzhuyu decoction.
