RESUMO
The decoction turns into a complex multiphase system following exposure to high temperature and a complex chemical environment. However, the differences in the concentration of key active ingredients in different phase states and the release of drugs in sedimentary phase have yet to be elucidated. A simple ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous quantitative determination of brucine, strychnine, liquiritin, isoliquiritin, isoliquiritigenin and glycyrrhizic acid concentrations and it was applied to compare the content of different phases and measure the release characteristics of the sedimentary phase in "Glycyrrhiza glabra-Nux vomica" decoction (NGD). The results show that the method's selectivity, precision (intraday and interday ≤ 2%), matrix effect (101-108%), recovery and stability results were acceptable according to the guidelines. The method is sensitive and reliable. The content determination results show that the most toxic strychnine in the sedimentary phase accounted for 75.70% of the total components. The different components exhibited differential release in different media, and its components were released in the artificial intestinal fluid up to 81.02% in 12 h. Several components conformed to the primary kinetic model and the Ritger-Peppas model, and the most toxic compound exhibited slow release, thus conforming to the Ritger-Peppas model. This study provides a standard of reference for studies investigating reduction in toxicity of the combination of Glycyrrhiza glabra (Glycyrrhiza glabra L.) and Nux vomica (Strychnos nux-vomica L.).
Assuntos
Strychnos nux-vomica , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Ácido Glicirrízico/análise , Sementes/química , Estricnina/química , Strychnos nux-vomica/química , Espectrometria de Massas em TandemRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Decoction is the most common form of administering traditional Chinese medicine (TCM). During the preparation of decoction, the high temperature and complex chemical environment result in the formation of complex and multiple phases. The differences in drug components in different phases induce gastrointestinal absorption and physiological response. Nux vomica (Strychnos nux-vomica L) is a typical toxic TCM used in China, with remarkable pharmacological activity. In order to reduce its toxicity, nux vomica (NV) is often decocted with Glycyrrhiza glabra (GG) in clinic, and the detoxification mechanism has always been the focus of research interest. Most studies investigated the compatibility of NV-GG, but the in vivo behavior of individual constituents based on phase state has yet to be elucidated. AIM OF THE STUDY: To investigate the pharmacokinetic behavior of typical toxic components in different phase states of "NV-GG decoction" in rat plasma. MATERIALS AND METHODS: The sediment, suspension, colloid and true solution of "NV-GG decoction" was obtained via physical methods. The main components in different phase states were analyzed via reliable UFLC-Q-TOF-MS high-resolution mass spectrometry. A rapid and accurate HPLC-qqq-MS/MS method was established and validated for accurate determination of brucine and strychnine levels in plasma, followed by pharmacokinetic evaluation of different phase states of "NV-GG decoction" in rats. Kinetex F5 100A (50 mm × 3.0 mm, 2.6 µm) column was used for chromatographic separation. Aqueous solution containing acetonitrile and 0.1% formic acid was used as the mobile phase, followed by gradient elution at 0.4 mL/min. Mass spectra were detected by electrospray ionization (ESI) multiple reaction monitoring (MRM) in positive ion mode. RESULTS: Fifteen different alkaloids were detected in different phase states of "NV-GG decoction". Strychnine and brucine, which are toxic components with high content, were selected for quantitative analysis. The established UPLC-qqq-MS/MS method is accurate and reliable with a good linearity (R2 > 0.99) in the respective concentration range, satisfying the quantitative requirements. The pharmacokinetic parameters of different phase states of rats differed significantly after gavage. The deposition phase was the most prominent. The index components showed higher Cmax, AUC0 and Tmax, while the T1/2, MRT, V/F and CL/F were the smallest, with a relatively slow plasma clearance rate in rats. The true solution group showed the lowest Tmax and the fastest absorption. CONCLUSION: This method has been successfully utilized to study the pharmacokinetics of different phase states of "NV-GG decoction". Among the four phases, the deposition phase contributed to a large proportion of the in vivo kinetic behavior similar to that of sustained-release preparations, with slow absorption of toxic components and prolonged peak time. The pharmacokinetic parameters and plasma concentration-time curves of each phase can be used to study toxicity reduction of NV-GG and increase its biocompatibility.
Assuntos
Alcaloides , Medicamentos de Ervas Chinesas , Glycyrrhiza , Strychnos nux-vomica , Administração Oral , Alcaloides/farmacocinética , Animais , Cromatografia Líquida de Alta Pressão , Ratos , Estricnina , Strychnos nux-vomica/química , Espectrometria de Massas em Tandem/métodosRESUMO
OBJECTIVES: In alternative medicine, plants pay a major role. Some plants are known for their poisonous nature but still have some importance in the herbal drug industry for their medicinal value. Strychnos nux-vomica is one such plant. Its nuts are called as poison nut due to the presence of alkaloids. Both the nut and its minerals are having medicinal properties and hence the present study was indented to understand the nature of primary metabolites and multi elemental composition. METHODS: The nuts of S. nux-vomica were procured, authenticated, powdered and subjected to proximate analysis parameters, visualization of thin layer chromatographic separation (TLC) and finger print profiling through high performance thin layer chromatographic (HPTLC); surface morphology by scanning electron microscopy, energy dispersive X-ray analysis, X-ray fluorescence spectrometry, X-ray photoelectron spectrometry, powder X-ray diffractometry and inductively coupled plasma optical emission spectrometry. RESULTS: In HPTLC, 7 spots each under 254 nm, 366 nm, derivatization with vanillin sulphuric acid (VSR) reagent appeared and 2 spots with Dragendorff's reagent. In HPTLC, 12 peaks at 254 nm, 9 peaks at 366 nm, 7 peaks at 520 nm after derivatization with VSR reagent detected. Elements such as potassium, calcium, magnesium, chlorine, aluminium, iron, manganese, sodium, nickel, phosphorus, copper, zinc, sulphur and silicon were identified. PXRD revealed that the presence of potassium chloride, calcite and dolomite as major elemental composition. CONCLUSIONS: The presence of all the above elements has vital roles on human physiology. Potassium, calcium, chlorine, aluminium, nickel, phosphorus, sulphur and silicon are reported for the first time in this study.
Assuntos
Strychnos nux-vomica , Alumínio/análise , Cálcio , Cloro , Humanos , Níquel , Nozes , Fósforo/análise , Potássio , Sementes/química , Silício/análise , Análise Espectral , Strychnos nux-vomica/química , Enxofre/análiseRESUMO
BACKGROUND: The establishment of strategy to inhibit the virus replication is an attractive means in combating SARS-CoV-2 infection. OBJECTIVE: We studied phyto-compounds from Strychnos nux-vomica (a poisonous plant) against SARS-CoV-2 RNA-dependent RNA polymerase by computational methods. METHODS: Molecular docking, molecular dynamics (MD) simulation and energetics calculations were employed to elucidate the role of the phyto-compounds. RESULTS: Ergotamine with a binding free energy of -14.39 kcal/mol showed a promising capability in terms of binding affinity and the interaction to conserved motifs, especially the SDD signature sequence. The calculated dissociation constants for ATP, ergotamine, isosungucine and sungucine were 12 µM, 0.072 nM, 0.011 nM and 0.152 nM, respectively. The exhibited kd by these phyto-compounds reflected tens of thousands fold potency as compared to ATP. The binding free energies of sungucine and isosungucine were much lower (-13.93 and -15.55 kcal/mol, respectively) compared to that of ATP (-6.98 kcal/mol). CONCLUSION: Sharing the same binding location as that of ATP and having high binding affinities, Ergotamine, Isosungucine, Sungucine and Strychnine N-oxide could be effective in controlling the SARS-CoV-2 virus replication by blocking the ATP and inhibiting the enzyme function.
Assuntos
COVID-19 , Strychnos nux-vomica , Trifosfato de Adenosina , Ergotaminas , Simulação de Acoplamento Molecular , Plantas Tóxicas , RNA Viral/genética , SARS-CoV-2 , Strychnos nux-vomica/químicaRESUMO
OBJECTIVES: We aimed to determine the circadian responses of mice to Semen Strychni and to investigate the role of pharmacokinetics in generating chronotoxicity. METHODS: Total extract of Semen Strychni was administered by oral gavage to wild-type (WT) and Bmal1-/- (a circadian clock-deficient model) mice at different circadian time points for toxicity (including survival) and pharmacokinetic characterization. Nephrotoxicity and neurotoxicity were evaluated by measuring plasma creatinine and creatine kinase BB (CK-BB), respectively. Drug metabolism and transport assays were performed using liver/intestine microsomes and everted gut sacs, respectively. KEY FINDINGS: Semen Strychni nephrotoxicity and neurotoxicity as well as animal survival displayed significant circadian rhythms (the highest level of toxicity was observed at ZT18 and the lowest level at ZT2 to ZT6). According to pharmacokinetic experiments, herb dosing at ZT18 generated higher plasma concentrations (and systemic exposure) of strychnine and brucine (two toxic constituents) compared with ZT6 dosing. This was accompanied by reduced formation of both dihydroxystrychnine and strychnine glucuronide (two strychnine metabolites) at ZT18. Bmal1 ablation sensitized mice to Semen Strychni-induced toxicity (with increased levels of plasma creatinine and CK-BB) and abolished the time dependency of toxicity. Metabolism of Semen Strychni (strychnine and brucine) in the liver and intestine microsomes of WT mice was more extensive at ZT6 than at ZT18. These time differences in hepatic and intestinal metabolism were lost in Bmal1-/- mice. Additionally, the intestinal efflux transport of Semen Strychni (strychnine and brucine) was more extensive at ZT6 than ZT18 in WT mice. However, the time-varying transport difference was abolished in Bmal1-/- mice. CONCLUSIONS: Circadian responses of mice to Semen Strychni are associated with time-varying efflux transport and metabolism regulated by the circadian clock (Bmal1). Our findings may have implications for optimizing phytotherapy with Semen Strychni via timed delivery.
Assuntos
Fatores de Transcrição ARNTL/genética , Ritmo Circadiano/fisiologia , Extratos Vegetais/toxicidade , Strychnos nux-vomica/química , Animais , Transporte Biológico , Relógios Circadianos/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microssomos/metabolismo , Síndromes Neurotóxicas/etiologia , Extratos Vegetais/administração & dosagem , Extratos Vegetais/farmacocinética , Estricnina/análogos & derivados , Estricnina/farmacocinética , Estricnina/toxicidade , Fatores de TempoRESUMO
Ethnopharmacological relevance Processed Nux vomica seed extracts and homeopathic medicinal preparations (HMPs) are widely used in traditional Indian and Chinese medicine for respiratory, digestive, neurological and behavioral disorders. Antioxidant property of Nux vomica is well known and recent investigation has highlighted the anticonvulsant potential of its homeopathic formulation. AIM OF THE STUDY: To explore the anticonvulsant and antiepileptogenic potential of Nux vomica HMPs (6CH, 12CH and 30CH potency) in pentylenetetrazole (PTZ) induced acute and chronic experimental seizure models in mice and investigate their effects on cognition, memory, motor activity and oxidative stress markers in kindled animals. MATERIALS AND METHODS: Acute seizures were induced in the animals through 70 mg/kg (i.p.) administration of PTZ followed by the evaluation of latency and duration of Generalized tonic-clonic seizures (GTCS). Subconvulsive PTZ doses (35 mg/kg, i.p.) induced kindling in 29 days, which was followed by assessment of cognition, memory and motor impairment through validated behavioral techniques. The status of oxidative stress was estimated through measurement of MDA, GSH and SOD. RESULTS: HMPs delayed the latency and reduced the duration of GTCS in acute model signifying possible regulation of GABAergic neurotransmission. Kindling was significantly hindered by the HMPs that justified the ameliorated cognition, memory and motor activity impairment. The HMPs attenuated lipid peroxidation by reducing MDA level and strengthened the antioxidant mechanism by enhancing the GSH and SOD levels in the kindled animals. CONCLUSIONS: Nux vomica HMPs showed anticonvulsant and antiepileptogenic potency in acute and chronic models of epilepsy. The test drugs attenuated behavioral impairment and reduced the oxidative stress against PTZ induced kindling owing to which they can be further explored for their cellular and molecular mechanism(s).
Assuntos
Anticonvulsivantes/farmacologia , Antioxidantes/farmacologia , Comportamento Animal/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Cognição/efeitos dos fármacos , Disfunção Cognitiva/prevenção & controle , Epilepsia/prevenção & controle , Transtornos da Memória/prevenção & controle , Memória/efeitos dos fármacos , Nootrópicos/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Strychnos nux-vomica , Doença Aguda , Animais , Anticonvulsivantes/isolamento & purificação , Antioxidantes/isolamento & purificação , Encéfalo/metabolismo , Encéfalo/fisiopatologia , Doença Crônica , Disfunção Cognitiva/etiologia , Disfunção Cognitiva/metabolismo , Disfunção Cognitiva/psicologia , Modelos Animais de Doenças , Epilepsia/induzido quimicamente , Epilepsia/metabolismo , Epilepsia/fisiopatologia , Excitação Neurológica/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Transtornos da Memória/etiologia , Transtornos da Memória/metabolismo , Transtornos da Memória/psicologia , Camundongos , Nootrópicos/isolamento & purificação , Pentilenotetrazol , Extratos Vegetais/isolamento & purificação , Strychnos nux-vomica/químicaRESUMO
OBJECTIVE: To investigate the antagonistic effect of the extract of Baizhu (Rhizoma Atractylodis Macrocephalae) (RAM) on the intestinal absorption of brucine and strychnine in Strychnos nux-vomica (NUX) and propose the mechanism of these effects. METHODS: The apparent permeability value (Papp) and absorption rate constant (Ka) were chosen as indices. The everted intestinal sac model and in situ single-pass intestinal perfusion model were used to study the effects of the RAM extract on the absorption of brucine and strychnine. To confirm the results, the brucine and strychnine concentrations in hepatic portal venous blood were determined. Western blotting was used to study P-glycoprotein (P-gp) expression in the Caco-2 cell line. RESULTS: Papp and Ka of brucine and strychnine were significantly increased in the presence of a P-gp inhibitor, but no significant increase was noted in the presence of a tight junction regulator. The RAM extract inhibited the absorption of brucine and strychnine and enhanced P-gp expression. CONCLUSION: The primary absorption mechanism for brucine and strychnine is passive transport, which is affected by P-gp.
Assuntos
Atractylodes/química , Medicamentos de Ervas Chinesas/farmacocinética , Absorção Intestinal/efeitos dos fármacos , Estricnina/análogos & derivados , Estricnina/farmacocinética , Strychnos nux-vomica/química , Animais , Células CACO-2 , Linhagem Celular Tumoral , Medicamentos de Ervas Chinesas/administração & dosagem , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Rizoma/química , Estricnina/administração & dosagemRESUMO
BACKGROUND: Peripheral nerve injury is a debilitating condition that may lead to partial or complete motor, sensory and autonomic function loss and lacks effective therapy until date. Therefore, it is quite imperative to explore impending remedies for rapid and accurate functional retrieval following such conditions. Natural product-based intervention can prove effective to facilitate the process of functions regain. METHODS: Here, we investigated the effect of processed Strychnos nux-vomica seeds at a dose of 250 mg/kg body weight in a mouse model of induced Sciatic nerve lesion in promoting the recovery of the functions. A compression injury was induced in the Sciatic nerve of the right leg in the mice. Sensory function recovery was evaluated by hot-plate and formalin tests, whereas the motor function retrieval was assessed by measuring muscle grip strength, sciatic functional index, and muscle mass restoration. Oxidative stress and blood cell count were measured by biochemistry and haematological analyses. RESULTS: This study indicates that Strychnos nux-vomica seeds enhance the rate of recovery of both sensory and motor functions. It helps restore the muscle mass, attenuates total oxidant status and enhances the total anti-oxidant capacity of the biological system. Moreover, the treated animals manifested an enhanced glucose tolerance aptitude and augmented granulocyte and platelet counts. Improved oxidant control, enhanced glucose sensitivity and amended granulocyte and platelet counts are likely to contribute to the advantageous effects of Strychnos nux-vomica, and warrant further in-depth studies for deciphering possible mechanisms and identification of active constituent(s) responsible for these effects. CONCLUSION: Strychnos nux-vomica seed offers functional recovery promoting effects following a mechanical injury to the Sciatic nerve and the possible reasons behind this effect can be reduced oxidative stress and improved glycaemic control. Further and detailed investigations can unravel this mystery.
Assuntos
Lesões por Esmagamento/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Traumatismos dos Nervos Periféricos/tratamento farmacológico , Preparações de Plantas/uso terapêutico , Neuropatia Ciática/tratamento farmacológico , Strychnos nux-vomica/química , Animais , Modelos Animais de Doenças , Masculino , Camundongos , Recuperação de Função Fisiológica , Sementes/químicaRESUMO
BACKGROUND Myasthenia gravis (MG) is a progressive autoimmune disorder caused by the production of antibodies directed against acetylcholine receptors (AChRs), resulting in muscle weakness and fatigue. This study aimed to explore the effect and mechanism of grilled nux vomica (GNV) in experimental autoimmune myasthenia gravis (EAMG) rats. MATERIAL AND METHODS Rat 97-116 peptides were used to mediate disease in the EAMG model in SPF female Lewis rats. The treatment groups received grilled nux vomica (75 mg/kg, 150 mg/kg, and 225 mg/kg). The autoantibody and inflammatory cytokines levels were measured by enzyme-linked immunosorbent assay (ELISA). RNA profiling was performed on high-dose and model group rats. Profiling results and TLR-4/NF-kappaB signaling were validated by q-PCR and Western blot analysis. RESULTS The results showed that GNV could attenuate the symptoms of EAMG rats. There was a decreased level of AChR-ab, IFN-γ, TNF-alpha, IL-2, IL-4, and IL-17 levels, and an increased level of TGF-ß1. In total, 235 differentially expressed genes (DEGs), consisting of 175 upregulated DEGs and 60 downregulated DEGs, were identified. Functional annotation demonstrated that DEGs were largely associated with leukocyte cell-cell adhesion, NF-kappa B signaling pathway, muscle contraction, and cardiac muscle contraction pathway. Rac2, Itgb2, Lcp2, Myl3, and Tnni1 were considered as hub genes with a higher degree value in the protein-protein interaction (PPI) network. The q-PCR and Western blot results of hub genes were consistent with RNA profiles. GNV treatment also significantly reduced the TLR-4 and NF-kappaB p65 protein expression in EAMG rats. CONCLUSIONS These results indicate that grilled nux vomica ameliorates EAMG by depressing the TLR-4/NF-kappaB signaling pathway, and hub genes may serve as potential targets for MG treatment.
Assuntos
Medicamentos de Ervas Chinesas/administração & dosagem , Miastenia Gravis Autoimune Experimental/tratamento farmacológico , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Strychnos nux-vomica/química , Animais , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Humanos , Músculo Esquelético/imunologia , Músculo Esquelético/patologia , Miastenia Gravis Autoimune Experimental/imunologia , Miastenia Gravis Autoimune Experimental/patologia , NF-kappa B/metabolismo , RNA-Seq , Ratos , Ratos Endogâmicos Lew , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Organismos Livres de Patógenos Específicos , Receptor 4 Toll-Like/metabolismoRESUMO
A simple, rapid, cost-effective and green analytical method is developed based on ultrasound-assisted dispersive liquid-liquid microextraction (US-DLLME) coupled to thin-layer chromatography (TLC)-image analysis for the simultaneous determination of two major alkaloids of Strychnos nux-vomica L i.e., strychnine and brucine. The method is composed of three steps, namely (i) US-DLLME by injecting a mixture of 100-µL chloroform (extraction solvent) and 1-mL methanol (disperser solvent) in 5 mL of aqueous sample, followed by ultrasonication and centrifugation, (ii) TLC of 20 µL of sedimented phase with methanol: ammonia (100:1.5, v/v) as the mobile phase and visualization under ultraviolet radiation (254 nm) and (iii) photography of TLC plate and quantification of spots by image analysis using freely available imageJ software (National Institute of Health, Bethesda, MD, USA). The limit of detection and limit of quantification for both alkaloids were found to be in the range of 0.12-0.15 and 0.36-0.48 µg/spot, respectively. The method was found to be linear in the range of 0.5-5 µg/spot with correlation coefficient (R2) of 0.995 and 0.997 for strychnine and brucine, respectively. The developed method was successfully applied for the determination of strychnine and brucine in Ayurvedic formulations and blood samples. The method does not require any sophisticated instrument and handling skills and can be adopted for rapid analysis of strychnine and brucine in forensic toxicological laboratories.
Assuntos
Cromatografia em Camada Fina/métodos , Microextração em Fase Líquida/métodos , Estricnina/análogos & derivados , Estricnina/análise , Strychnos nux-vomica/química , Cromatografia em Camada Fina/economia , Análise Custo-Benefício , Humanos , Processamento de Imagem Assistida por Computador , Limite de Detecção , Microextração em Fase Líquida/economia , Ayurveda , Reprodutibilidade dos Testes , Estricnina/sangue , Comprimidos/análise , Ultrassom , Raios UltravioletaRESUMO
The ripened seeds of Strychnos nux-vomica L. have been extensively used as herbal medicines in Asian countries. Dihydroindole-type alkaloids are not only the active constituents but also the toxicants in Strychnos. However, the simultaneous determination of these alkaloids in both crude and processed Semen Strychni is still lacking. The present study represents the first quantitation and relative quantitation assay of 12 dihydroindole-type alkaloids in Strychnos nux-vomica unprocessed and sand-processed seeds using high-performance liquid chromatography coupled with diode array detection and mass spectrometry. The relative concentration of ten alkaloids was calculated by semi-quantification using the internal standard and their amounts in unprocessed and detoxified Semen Strychni were compared. We report here for the first time the significant increase of the two alkaloids, 19-N-methyl-strychnine, and 2,3-dimethoxy-19-N-methyl-strychnine, during the processing of Semen Strychni. Our study provides new insight into the true complexity of seed processing procedure and valuable information for assessing the efficacy and safety for clinical applications of Semen Strychni-containing drugs.
Assuntos
Alcaloides Indólicos/análise , Sementes/química , Strychnos nux-vomica/química , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas , Estrutura MolecularRESUMO
Brucine and Strychnine are alkaloids isolated from the seeds of Strychnos nux vomica L., which have long been used as a traditional medicine for the treatment of tumor. However, the effect of Brucine and Strychnine on colorectal cancer (CRC) and the underlying molecular mechanism remain unclear. In the present study, Brucine and Strychnine displayed profound inhibitory effects on the growth of human colon cancer cells. The results of flow cytometric analysis demonstrated that the two alkaloids induced cellular apoptosis. Moreover, the growth of DLD1 xenografted tumors in nude mice was significantly suppressed in the Brucine or Strychnine treated group. Mechanistically, the Wnt/ß-catenin is involved in this phenomenon, which is characterized by significantly increased expression of DKK1 and APC, whereas decreased expression of ß-catenin, c-Myc, and p-LRP6 in CRC cells as well as tumor tissues. Collectively, Brucine and Strychnine have targeted inhibition for colon cancer proliferation both in vitro and in vivo, and it is valuable for future exploitation and utilization as an antitumor agent of CRC.
Assuntos
Alcaloides/química , Neoplasias do Colo/tratamento farmacológico , Estricnina/análogos & derivados , Estricnina/química , Strychnos nux-vomica/química , Via de Sinalização Wnt/efeitos dos fármacos , Animais , Neoplasias do Colo/patologia , Humanos , Camundongos , Camundongos NusRESUMO
Vasculogenic mimicry (VM) with the pattern of endothelial independent tubular structure formation lined by aggressive tumor cells mimics regular tumor blood vessels to ensure robust blood supply and correlates with the proliferation, invasion, metastasis, and poor prognosis of malignant tumors, which was demonstrated to be a major obstacle for resistance to antiangiogenesis therapy. Therefore, it is urgent to discover methods to abrogate the VM formation of tumors, which possesses important practical significance for improving tumor therapy. Brucine is a traditional medicinal herb extracted from seeds of Strychnos nux-vomica L. (Loganiaceae) exhibiting antitumor activity in a variety of cancer models. In the present study, the effect of brucine on vasculogenic mimicry and the related mechanism are to be investigated. We demonstrated that, in a triple-negative breast cancer cell line MDA-MB-231, brucine induced a dose-dependent inhibitory effect on cell proliferation along with apoptosis induction at higher concentrations. The further study showed that brucine inhibited cell migration and invasion with a dose-dependent manner. Our results for the first time indicated that brucine could disrupt F-actin cytoskeleton and microtubule structure, thereby impairing hallmarks of aggressive tumors, like migration, invasion, and holding a possibility of suppressing vasculogenic mimicry. Hence, the inhibitory effect of brucine on vasculogenic mimicry was further verified. The results illustrated that brucine significantly suppressed vasculogenic mimicry tube formation with a dose-dependent effect indicated by the change of the number of tubules, intersections, and mean length of tubules. The in-depth molecular mechanism of vasculogenic mimicry suppression induced by brucine was finally suggested. It was demonstrated that brucine inhibited vasculogenic mimicry which might be through the downregulation of erythropoietin-producing hepatocellular carcinoma-A2 and matrix metalloproteinase-2 and metalloproteinase-9.
Assuntos
Neovascularização Patológica/tratamento farmacológico , Estricnina/análogos & derivados , Strychnos nux-vomica/química , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Neovascularização Patológica/genética , Neovascularização Patológica/patologia , Estricnina/química , Estricnina/farmacologia , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/patologiaRESUMO
Nux vomica has been effectively used in Traditional Chinese Medicine. The processing of Nux vomica is necessary to reduce toxicity before it can be used in clinical practice. However, the mechanism for processing detoxification is unclear. hERG channels have been subjected to a routine test for compound cardiac toxicity in the drug development process. Therefore, we examined the effects and mechanisms of strychnine and brucine, two main ingredients of Nux vomica, and their N-oxides on hERG channels. Strychnine and brucine exhibited concentration-dependent inhibition of hERG channels with IC50 values of 25.9⯵M and 44.18⯵M, respectively. However, their nitrogen oxidative derivatives produced by processing of Nux vomica, strychnine N-oxide and brucine N-oxide, lost their activity on hERG channels. Compared to their parent compounds, only an oxygen atom was introduced in the nitrogen oxidative isoforms to compensate for the N+â¯-â¯charge, suggesting that the protonated nitrogen is the key group for strychnine and brucine binding to hERG channel. Alanine-mutagenesis identified Y652 is the most important residue for strychnine and brucine binding to hERG channel. Y652A mutation increased the IC50 for strychnine and brucine by 21.64-fold and 29.78-fold that of WT IhERG, respectively. Docking simulations suggested that the protonated nitrogen of strychnine and brucine formed a cation-π interaction with the aromatic ring of Y652. This study suggests that introduction of an oxygen to compensate for the N+â¯-â¯charge could be a useful strategy for reducing hERG potency and increasing the safety margin of alkaloid-type compounds in drug development.
Assuntos
Oxigênio/metabolismo , Canais de Potássio/metabolismo , Estricnina/análogos & derivados , Estricnina/metabolismo , Alcaloides/metabolismo , Linhagem Celular , Células HEK293 , Humanos , Medicina Tradicional Chinesa/métodos , Nitrogênio/metabolismo , Sódio/metabolismo , Relação Estrutura-Atividade , Strychnos nux-vomica/química , Regulador Transcricional ERG/metabolismoRESUMO
Strychnos nux-vomica L. belongs to the genus Strychnos of the family Loganiaceae and grows in Sri Lanka, India and Australia. The traditional medicinal component is its seed, called Nux vomica. This study provides a relevant and comprehensive review of S. nux-vomica L., including its botany, ethnopharmacology, phytochemistry, pharmacology and toxicology, thus providing a foundation for future studies. Up to the present day, over 84 compounds, including alkaloids, iridoid glycosides, flavonoid glycosides, triterpenoids, steroids and organic acids, among others, have been isolated and identified from S. nux-vomica. These compounds possess an array of biological activities, including effects on the nervous system, analgesic and anti-inflammatory actions, antitumor effects, inhibition of the growth of pathogenic microorganisms and regulation of immune function. Furthermore, toxicity and detoxification methods are preliminarily discussed toward the end of this review. In further research on S. nux-vomica, bioactivity-guided isolation strategies should be emphasized. Its antitumor effects should be investigated further and in vivo animal experiments should be performed alongside in vitro testing. The pharmacological activity and toxicology of strychnine [Formula: see text]-oxide and brucine [Formula: see text]-oxide should be studied to explore the detoxification mechanism associated with processing more deeply.
Assuntos
Strychnos nux-vomica/química , Strychnos nux-vomica/toxicidade , Alcaloides/isolamento & purificação , Alcaloides/farmacologia , Analgésicos , Animais , Anti-Inflamatórios , Antineoplásicos Fitogênicos , Óxidos N-Cíclicos/farmacologia , Óxidos N-Cíclicos/toxicidade , Flavonoides/isolamento & purificação , Flavonoides/farmacologia , Humanos , Técnicas In Vitro , Glicosídeos Iridoides/isolamento & purificação , Glicosídeos Iridoides/farmacologia , Loganiaceae , Fitoterapia , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Sementes/química , Estricnina/análogos & derivados , Estricnina/farmacologia , Estricnina/toxicidadeRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: The dried ripe seeds of Nux Vomica (Strychnos nux-vomica L.), a traditional Chinese medicine, have been used to treat multifarious symptoms. However, the clinical applications of Nux Vomica are limited by its severe toxicity. In this study, Nux Vomica was subjected to nuclear magnetic resonance (NMR) metabonomics and pathological examination to determine relevant biomarkers in target organs and to explain the underlying toxicity mechanism. MATERIALS AND METHOD: Thirty-six male Sprague-Dawley rats were randomly divided into three groups of twelve rats. The control group was oral gavaged with distilled water, and two experiment groups were treated with Nux Vomica at a dose of 0.315 and 0.630g/kg body weight. On days 14 and 21, serum, urine, liver and kidney tissues were collected for histopathological examination, biochemical analysis and 1H-NMR analysis. RESULTS: The metabolites changes of rats treated with Nux Vomica are obviously differ from that of controls. In serum, low-dose group compared with control shows the significantly changes included elevated concentration of glucose, TMAO, and creatine, with decreased lipids, 3-HB, lactate, and unsaturated fatty acid. Change in taurine was only observed in the separation comparison of high-dose group and control. In urine, the variation metabolites included elevations in glucose, creatine, and TMAO as well as decreased lactate, succinate, α-ketoglutaric acid, citrate and hippurate in low-dose group compared with control. Only alanine and creatine were decreased significantly in high-dose group compared with control. CONCLUSION: Nux Vomica induced disruptions in glycolysis, lipid and amino acid metabolism, and toxic effects were aggravated in liver and kidney tissues as dosing time was prolonged. 1H NMR-based metabonomics combined with biochemical and histopathological methods can be applied to elucidate the toxicity mechanism of Nux Vomica decoction that caused liver and kidney injuries in rats.
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Medicamentos de Ervas Chinesas/toxicidade , Metabolômica/métodos , Extratos Vegetais/toxicidade , Strychnos nux-vomica/química , Aminoácidos/efeitos dos fármacos , Aminoácidos/metabolismo , Animais , Química Clínica/métodos , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/administração & dosagem , Glicólise/efeitos dos fármacos , Rim/efeitos dos fármacos , Rim/patologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Medicina Tradicional Chinesa/efeitos adversos , Extratos Vegetais/administração & dosagem , Espectroscopia de Prótons por Ressonância Magnética , Ratos , Ratos Sprague-Dawley , Sementes , Fatores de TempoRESUMO
In this study, we have aimed to analyze the phytochemical composition of this plant and the concentration of strychnine and brucine. The identification of bioactive compounds was done by GC-MS with NIST Library. Strychnine and Brucine were quantified using HPLC. Twenty one medicinal bio active compounds were identified from the Strychnos nux-vomica leaf ethanolic extract. Strychnine is showing 28.43% purity and brucine was not detected in GCMS analysis. Quantified the concentration of strychnine (0.6 mg in 500mg of extract) and brucine (1.6 mg in 500mg of extract) was done by HPLC against Strychnine and Brucine standard. These compounds are having natural properties of Anti-inflammatory, Hypocholesterole, Cancer preventive, Hepatoprotective, Antimicrobial, Antioxidant, Cardio protective, Antiaging, Antialzheimeran, Antidermatitic, Immunostimulant, Anthepatotoxic, biosynthesis of steroid hormones, Nematicide, Antiandrogenic, 5-alpha reductase inhibitor, antipsychotic, analgesic, apoptotic effect, antidepressant, antidote for snake poisoning and diabetic activity.
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Cromatografia Líquida de Alta Pressão , Cromatografia Gasosa-Espectrometria de Massas , Extratos Vegetais/análise , Extratos Vegetais/farmacologia , Folhas de Planta/química , Estricnina/análogos & derivados , Strychnos nux-vomica/química , Estricnina/análise , Estricnina/farmacologiaRESUMO
The seeds of Strychnosnux-vomica L., as a traditional Chinese medicine, have good anti-inflammatory and analgesic activities. However, it usually leads to gastrointestinal irritation and systemic toxicity via oral administration. In the study, it was discovered that a novel gel transdermal delivery system contained brucine, the main effective component extracted from Strychnosnux-vomica. Results showed that the brucine gel system inhibited arthritis symptoms and the proliferation of the synoviocytes in the rat adjuvant arthritis model, which indicated its curative effect for rheumatoid arthritis. Meanwhile, it significantly relieved the xylene-induced ear edema in the mouse ear swelling test, which manifested its anti-inflammatory property. Moreover, the brucine gel eased the pain of paw formalin injection in the formalin test, which demonstrated its analgesic effects. In addition, the brucine significantly inhibited lipopolysaccharide (LPS)-induced Prostaglandin E2 (PGE2) production without affecting the viability of cell in vitro anti-inflammatory test, which proved that its anti-inflammatory and analgesic actions were related to inhibition of prostaglandin synthesis. It is suggested that the brucine gel is a promising vehicle for transdermal delivery on the treatment of inflammatory disease.
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Analgésicos/farmacologia , Anti-Inflamatórios/farmacologia , Modelos Animais de Doenças , Estricnina/análogos & derivados , Administração Cutânea , Analgésicos/administração & dosagem , Animais , Anti-Inflamatórios/administração & dosagem , Artrite Experimental/patologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Dinoprostona/biossíntese , Sistemas de Liberação de Medicamentos/métodos , Edema/prevenção & controle , Formaldeído , Géis , Humanos , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Dor/induzido quimicamente , Dor/prevenção & controle , Fitoterapia , Ratos Wistar , Estricnina/administração & dosagem , Estricnina/farmacologia , Strychnos nux-vomica/química , Sinoviócitos/efeitos dos fármacos , Sinoviócitos/patologiaRESUMO
INTRODUCTION: ß-Tubulin is an important target for the binding of anti-cancer drugs, in particular, paclitaxel (taxol), vinblastine and epothilone. However, mutations in ß-tubulin structure give resistance to chemotherapeutic agents. Notably, mutations at R306C, F270 V, L217R, L228F, A185T and A248V positions in ß-tubulin give high resistance for paclitaxel binding. OBJECTIVE: To discover novel inhibitors of ß-tubulin from natural sources, particularly alkaloids, using a virtual screening approach. METHODOLOGY: A virtual screening approach was employed to find potent lead molecules from the Naturally-occurring Plant-based Anti-cancer Compound-activity Target (NPACT) database. Alkaloids have great potential to be anti-cancer agents. Therefore, we have screened all alkaloids from a total of 1574 molecules from the NPACT database for our study. Initially, Molinspiration and DataWarrior programs were utilised to calculate pharmacokinetics and toxicity risks of the alkaloids, respectively. Subsequently, AutoDock algorithm was employed to understand the binding efficiency of alkaloids against ß-tubulin. The binding affinity of the docked complex was confirmed by means of an intermolecular interaction study. Moreover, oral toxicity was predicted by using ProTox program. Further, metabolising capacity of drugs was studied by using SmartCYP software. Additionally, scaffold analysis was done with the help of scaffold trees and dendrograms, providing knowledge about the building blocks for parent-compound synthesis. RESULTS: Overall, the results of our computational analysis indicate that isostrychnine, obtained from Strychnosnux-vomica, satisfies pharmacokinetic and bioavailability properties, binds efficiently with ß-tubulin. Thus, it could be a promising lead for the treatment of paclitaxel resistant cancer types. CONCLUSION: This is the first observation of inhibitory activity of isostrychnine against ß-tubulin and warrants further experimental investigation. Copyright © 2016 John Wiley & Sons, Ltd.
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Alcaloides/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Compostos Fitoquímicos/farmacologia , Moduladores de Tubulina/química , Moduladores de Tubulina/farmacologia , Administração Oral , Disponibilidade Biológica , Bases de Dados de Compostos Químicos , Humanos , Simulação de Acoplamento Molecular , Compostos Fitoquímicos/química , Plantas/química , Estricnina/química , Estricnina/farmacologia , Strychnos nux-vomica/química , Testes de Toxicidade , Interface Usuário-ComputadorRESUMO
Brucine is the active component in traditional Chinese medicine "Ma-Qian-Zi" (Strychnos nux-vomica Linn), with capabilities of analgesic, anti-inflammatory, anti-tumor and so on. It is crucial how to break through the impact of cuticle skin which reduces the penetration of drugs to improve drug transmission rate. The aim of this study is to improve the local drug concentration by using ultrasound. We used fresh porcine skin to study the effects of ultrasound on the transdermal absorption of brucine under the influence of various acoustic parameters, including frequency, amplitude and irradiation time. The transdermal conditions of yellow-green fluorescent nanoparticles and brucine in skin samples were observed by laser confocal microscopy and ultraviolet spectrophotometry. The results show that under ultrasonic conditions, the permeability of the skin to the fluorescent label and brucine (e.g., the depth and concentration of penetration) is increased compared to its passive diffusion permeability. The best ultrasound penetration can make the penetration depth of more than 110 microns, fluorescent nanoparticles and brucine concentration increased to 2-3 times. This work will provide supportive data on how the brucine is better used for transdermal drug delivery (TDD).