Assuntos
Medicamentos de Ervas Chinesas , Farmácias , Farmácia , Humanos , Medicina Tradicional Chinesa , TaiwanRESUMO
This study investigated the antifatigue effects of rutin, a flavonoid extracted from the ethyl acetate extract of S. involucrata. Mice were subjected to a weight-loaded forced swim test (WFST) on alternate days for 3 wk. Rutin was administered orally to the mice for 7 days in dosages of 15, 30, and 60 mg/kg body weight, and several biomarkers of physical fatigue were evaluated: swimming time, change in body weight, lipid peroxidation, lactic acid (LA), glycogen, and the activities of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx). On Day 7, the rutin-treated mice had a 3-fold longer exhaustive swimming time than the control mice, as well as significantly reduced blood LA concentrations. The 15, 30, and 60 mg/kg body weight rutin-supplemented groups displayed 11.2%, 22.5%, and 37.7% reduced malondialdehyde (MDA) concentrations, respectively, in brain and muscle tissues compared with the control exercised group. Our results indicated that the administration of rutin protected the mice against the depletion of SOD and GPx activities significantly. Following 7 days of rutin treatment, we sacrificed the mice and analyzed their soleus muscle and brain for peroxisome proliferator-activated receptor-α coactivator (PGC-1α) and sirtuin 1 (SIRT1) mRNA expression. We observed that rutin treatment increased PGC-1α and SIRT1 mRNA and protein expression. The changes in these markers of mitochondrial biogenesis were associated with increased maximal endurance capacity. The application of 2D gel electrophoresis to analyze the rutin-responsive protein profiles in the WFST mouse brain further revealed the upregulation of the CB1 cannabinoid receptor-interacting protein 1, myelin basic protein, Rho GDP dissociation inhibitor (GDI) alpha, and TPI, indicating that rutin might inhibit anxiety through the upregulation of the expression of anxiety-associated proteins. Western blot analysis of MAPK expression further confirmed the antianxiety effects of rutin. Our study results thus indicate that rutin treatment ameliorates the various impairments associated with physical fatigue.
Assuntos
Antioxidantes/metabolismo , Fadiga/tratamento farmacológico , Rutina/administração & dosagem , Saussurea/química , Animais , Peso Corporal/efeitos dos fármacos , Fadiga/patologia , Glutationa Peroxidase/biossíntese , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Camundongos , Condicionamento Físico Animal , Rutina/química , Superóxido Dismutase/biossíntese , NataçãoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Liver fibrosis is the result of long-term liver damage and the wound-healing process, in which the hepatic stellate cell (HSC) plays a crucial role during fibrogenesis. The liver sinusoidal endothelial cell (LSEC) is a liver-resident scavenger, contributing to sinusoidal remodeling, HSC activation and liver fibrosis. Lipopolysaccharide (LPS) causes an inflammatory reaction associated with portal circulation and LSECs signaling. Scutellariae radix, the root of Scutellaria baicalensis Georgi, is a Chinese herb widely used for liver diseases. However, its effect on LSEC activation and HSC migration in liver fibrosis has not been investigated yet. AIM OF THIS STUDY: LPS-induced rat LSEC (rLSEC) activation was used as a model to screen and explore the active components of Scutellariae radix. The anti-fibrotic effect of Scutellariae radix on rLSEC activation and rHSC migration was further investigated. MATERIALS AND METHODS: LPS-induced rLSEC mRNA expression, including VEGF, VEGFR, MCP-1, and TGF-ß1, were examined by real-time PCR analyses. MCP-1 protein levels were measured by an ELISA kit. rLSEC conditioned medium on rHSC migration was measured by wound-healing assay and transwell chemoattraction assay. RESULTS: Results showed LPS-induced rLSEC activation with upregulated MCP-1 mRNA and protein expressions, and that rLSEC-condition medium enhanced rHSC migration. Both baicalein and wogonin from the active subfraction significantly reduced MCP-1 expression, but only baicalein markedly inhibited rHSC migration in rLSEC conditioned medium. CONCLUSION: This study demonstrated that Scutellariae radix attenuates LPS-induced rLSEC activation and HSC migration with downregulation of MCP-1 expression. The results provide supporting evidence that Scutellariae radix may be beneficial for the amelioration of liver fibrosis.
Assuntos
Movimento Celular/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Flavonoides/farmacologia , Células Estreladas do Fígado/efeitos dos fármacos , Extratos Vegetais/farmacologia , Scutellaria baicalensis , Animais , Movimento Celular/fisiologia , Quimiocina CCL2/genética , Células Endoteliais/fisiologia , Regulação da Expressão Gênica/efeitos dos fármacos , Células Estreladas do Fígado/fisiologia , Lipopolissacarídeos , Fígado/citologia , Raízes de Plantas , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de Fatores de Crescimento do Endotélio Vascular/genética , Fator de Crescimento Transformador beta1/genética , Fator A de Crescimento do Endotélio Vascular/genética , CicatrizaçãoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: GP-TCM is the first EU-funded Coordination Action consortium dedicated to traditional Chinese medicine (TCM) research. One of the key deliverables of the Work Package 7 in GP-TCM was to investigate information of the existing requirements for registration of TCM products listed by global regulatory bodies. The paper aims to collate data and draw comparison of these regulations. Case studies are also presented to illustrate the problems involved in registering TCM products in different regions worldwide. MATERIALS AND METHODS: A collaborative network task force was established during the early stage of the GP-TCM project and operated through exchanges, teleconferences and focused discussions at annual meetings. The task force involved coordinators, academics who are actively involved with R&D of Chinese herbal medicines, experts on monographic standards of Chinese materia medica, representatives from regulatory agencies, experts from industries in marketing Chinese medicines/herbal medicines and natural products. The co-ordinators took turns to chair teleconferences, led discussions on specific issues at AGM discussion sessions, at joint workshops with other work-packages such as WP1 (quality issues), WP3 (toxicology issues) and WP6 (clinical trial issues). Collectively the authors were responsible for collating discussion outcomes and updating written information. RESULTS: A global overview of regulations on herbal registration has been compiled during the three years of the consortium. The regulatory requirements for registration of herbal products in the EU and China were compared, and this is extended to other regions/countries: Africa, Australia, Brazil, Canada, Japan, Russia, South Korea, Taiwan, and the United States. A wide variation of the regulations for the categories of herbal products exists: food (functional food, novel foods, dietary food for special medical purpose, foods for particular nutritional use, food supplement); cosmetic, traditional herbal medicine products; herbal medicines for human use and veterinary use. CONCLUSION: The regulatory issues for registration of herbal products are complicated among the countries and regions worldwide. The information summarised in the text is for reference only. Some regulations which are presented in this review are still in legislation process and may change in due course. Before taking any regulatory action, readers are advised to consult current official legislation and guidance and/or to seek appropriate professional advice. The lessons learnt from global regulation of TCM will provide valuable insights for regulation of other traditional medicine such as Ayurveda and Unani medicine, as well as other forms of indigenous medicine. The WHO is well placed to co-ordinate a consultation process with the aim of putting forward suggestions for harmonisation to key regulatory agencies.
Assuntos
Qualidade de Produtos para o Consumidor/legislação & jurisprudência , Medicamentos de Ervas Chinesas , Regulamentação Governamental , Legislação de Medicamentos , Legislação sobre Alimentos , Medicina Tradicional Chinesa , Fitoterapia , África , Ásia , Austrália , Brasil , China , União Europeia , Previsões , Humanos , Internacionalidade , Plantas Medicinais , Estados UnidosRESUMO
Hepatic stellate cells (HSCs) play a key role in the pathogenesis of liver fibrosis. In chronic liver injury, HSCs undergo transdifferentiation to an activated myofibroblastic phenotype and migrate to injured areas in response to chemotactic factors, producing extracellular matrix proteins such as collagen type I to repair the damage as well as overexpression of α-smooth muscle actin (α-SMA). Paeoniae Radix, the root of Paeonia lactiflora Pall, was investigated for PDGF-BB-induced HSC chemotaxis. Rat HSCs and LX-2, a human HSC cell line, were used for the in vitro experiments. Cell migration was analyzed by wound-healing and transwell assays. An ELISA and a Sircol collagen assay kit were used to detect the expressions of α-SMA and of collagen, respectively. Phosphorylations of mitogen-activated protein kinases, including ERK 1/2, p38, and JNK, were evaluated with immunoblotting. Results indicated that PDGF-BB increased migration as well as α-SMA and collagen expression in HSCs. Paeoniae Radix extracts and its active components, paeonol and 1,2,3,4,6-penta- O-galloyl- ß-D-glucose (PGG), inhibited PDGF-BB-induced HSC migration and α-SMA and collagen expressions in a concentration-dependent manner. The inhibitory effects were associated with downregulation of PDGF receptor- α, ERK, p38, and JNK activation. Both paeonol and PGG participate in HSC migration, but via differential mechanisms.
Assuntos
Movimento Celular/efeitos dos fármacos , Células Estreladas do Fígado/efeitos dos fármacos , Paeonia/química , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas c-sis/antagonistas & inibidores , Acetofenonas/farmacologia , Actinas/antagonistas & inibidores , Actinas/biossíntese , Animais , Becaplermina , Linhagem Celular , Transdiferenciação Celular/efeitos dos fármacos , Quimiotaxia/efeitos dos fármacos , Colágeno/antagonistas & inibidores , Colágeno/biossíntese , Ensaio de Imunoadsorção Enzimática , Células Estreladas do Fígado/citologia , Células Estreladas do Fígado/metabolismo , Humanos , Taninos Hidrolisáveis/farmacologia , Masculino , Raízes de Plantas/química , Ratos , Ratos Sprague-DawleyRESUMO
The aim of this study was to investigate if armepavine (Arm, C19H23O3N) could exert inhibitory effects against hepatic fibrosis in rats. A cell line of rat hepatic stellate cells (HSC-T6) was stimulated with tumour necrosis factor-α (TNF-α) to evaluate the inhibitory effects of Arm. Rats were injected with thioacetamide (TAA; 300 mg/kg, intraperitoneally) thrice a week for 4 weeks to induce hepatic fibrosis, with Arm (3 or 10 mg/kg) given by gavage twice a day. Liver sections were taken for western blotting, fibrosis scoring and immunofluorescence staining. Arm (1-10 µm) concentration-dependently attenuated TNF-α-stimulated: (i) protein expressions of α-smooth muscle actin (α-SMA), collagen type I and angiopoietin-1; (ii) H2O2 production; and (iii) NF-κB, JunD and C/EBPß (cytidine-cytidine-adenosine-adenosine-thymidine (CCAAT)/enhancer binding protein-ß (EBPß)) nuclear translocations in HSC-T6 cells. In vivo Arm treatment significantly reduced plasma aspartate transaminase and alanine transaminase levels, hepatic α-SMA expression and collagen contents, and fibrosis scores of TAA-injected rats. Moreover, Arm treatment decreased α-SMA- and NF-κB-positive cells in immunohistochemical staining, and mRNA expression levels of IL-6, TGF-ß1, TIMP-1, col1α2, iNOS and ICAM-1 genes, but up-regulated the metallothionein gene in the livers of TAA-injected rats. Our results indicated that Arm exerted both in vitro and in vivo antifibrotic effects in rats, with inhibition of NF-κB, JunD and C/EBPß pathways.
Assuntos
Benzilisoquinolinas/uso terapêutico , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/tratamento farmacológico , Fitoterapia , Tioacetamida/efeitos adversos , Actinas/genética , Actinas/metabolismo , Transporte Ativo do Núcleo Celular , Alanina Transaminase/sangue , Angiopoietina-1/genética , Angiopoietina-1/metabolismo , Animais , Aspartato Aminotransferases/sangue , Benzilisoquinolinas/administração & dosagem , Western Blotting , Proteína beta Intensificadora de Ligação a CCAAT/genética , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Colágeno Tipo I/metabolismo , Citocinas/genética , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Imunofluorescência , Células Estreladas do Fígado/citologia , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Peróxido de Hidrogênio/metabolismo , Cirrose Hepática/patologia , Masculino , Metalotioneína/genética , Metalotioneína/metabolismo , NF-kappa B/metabolismo , Ratos , Ratos Sprague-Dawley , Tioacetamida/administração & dosagem , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Transcrição GênicaRESUMO
BACKGROUND: We previously demonstrated that kaerophyllin, a lignan, isolated from a widely used traditional Chinese herb, Bupleurum scorzonerifolium, leading to the inhibition of hepatic stellate cells (HSCs) activation in vitro. This current study evaluated the in vivo role of kaerophyllin in protecting the liver against injury and fibrogenesis caused by thioacetamide (TAA) in rats and further explored the underlying mechanisms. MATERIALS AND METHODS: Liver fibrosis in Sprague-Dawley rats was induced by intraperitoneal injection of TAA (200 mg/kg) twice per week for 6 weeks. Animals were divided into five groups: vehicle control, TAA control, TAA + low dose kaerophyllin, TAA + high dose kaerophyllin and TAA + curcumin groups. Kaerophyllin (10 or 30 mg/kg) or curcumin (150 mg/mL) was given by gavage twice per day consecutively for 4 weeks starting 2 weeks after TAA injection. Rat HSCs were used to investigate the anti-inflammatory role of kaerophyllin against tumour necrosis factor α (TNF-α) in vitro. Peroxisome proliferator-activated receptor-γ (PPAR-γ) expression was knocked down in rat HSCs using PPAR-γ small interfering RNAs. RESULTS: Kaerophyllin significantly protected liver from injury by reducing serum aspartate transaminase and alanine transaminase levels and by improving the histological architecture and fibrosis score. In addition, kaerophyllin suppressed inflammation by reducing the mRNA of TNF-α, interleukin-1ß (IL-1ß) and monocyte chemoattractant protein-1 (MCP-1) genes. In HSCs, kaerophyllin elevated PPAR-γ activity and reduced TNF-α-stimulated mRNA levels of intracellular adhesion molecule-1 (ICAM-1), MCP-1 and IL-1ß genes, which were reversed by small interfering RNA knockdown of PPAR-γ gene. CONCLUSIONS: Our results demonstrated that kaerophyllin protected the rat liver from TAA-caused injury and fibrogenesis by suppressing hepatic inflammation and inhibiting HSC activation, possibly through upregulation of PPAR-γ expression.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Células Estreladas do Fígado/efeitos dos fármacos , Lignanas/uso terapêutico , Cirrose Hepática/prevenção & controle , Fígado/efeitos dos fármacos , Tioacetamida/efeitos adversos , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Carcinógenos/metabolismo , Células Estreladas do Fígado/metabolismo , Fígado/metabolismo , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/tratamento farmacológico , Masculino , Extratos Vegetais/metabolismo , Extratos Vegetais/uso terapêutico , Raízes de Plantas , Ratos , Ratos Sprague-Dawley , Tioacetamida/metabolismoRESUMO
BACKGROUND: Hepatic stellate cells (HSCs), the key cell type for hepatic fibrosis, become activated and profibrogenic in the presence of hepatocyte apoptotic bodies (ABs). Bupleurum scorzonerifolium (BS), a widely used traditional Chinese herb for liver diseases, was fractionated, and the inhibitory effects of BS extracts on AB-induced HSC migration were screened. The activity-guided fractionation led to a lignan, kaerophyllin. In this study, the anti-fibrotic effects of kaerophyllin were studied in the presence of ABs. METHODS: LX-2 cells phagocytosing ultraviolet (UV)-induced HepG2 ABs were investigated by confocal microscopy and flow cytometry. AB-induced HSC activation was evaluated by immunoblotting and real-time PCR analyses. HSC migration was measured by wound-healing assays. RESULTS: HepG2 ABs induced LX-2 activation, with the production of collagen I and α-smooth muscle actin, upregulated profibrogenic gene transcriptions and increased NF-κB activity, cell migration and phagocytosis. Kaerophyllin from BS antagonized AB-induced HSC migration and activation. CONCLUSIONS: Kaerophyllin inhibited AB-induced LX-2 activation and migration with downregulation of Akt/ERK phosphorylations and NF-κB activity. Our study suggests a novel platform for screening anti-fibrotic compounds with ABs.
Assuntos
Apoptose/efeitos dos fármacos , Bupleurum , Células Estreladas do Fígado/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Lignanas/farmacologia , Extratos Vegetais , Actinas/metabolismo , Apoptose/efeitos da radiação , Western Blotting , Bupleurum/química , Movimento Celular/efeitos dos fármacos , Colágeno Tipo I/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fibrose , Citometria de Fluxo , Regulação da Expressão Gênica , Células Hep G2 , Células Estreladas do Fígado/metabolismo , Células Estreladas do Fígado/patologia , Hepatócitos/patologia , Hepatócitos/efeitos da radiação , Humanos , Lignanas/isolamento & purificação , Microscopia Confocal , Quinase de Cadeia Leve de Miosina/antagonistas & inibidores , Quinase de Cadeia Leve de Miosina/metabolismo , NF-kappa B/metabolismo , Proteína Oncogênica v-akt/antagonistas & inibidores , Proteína Oncogênica v-akt/metabolismo , Fagocitose , Extratos Vegetais/química , Inibidores de Proteínas Quinases/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Raios UltravioletaRESUMO
Platelet-derived growth factor (PDGF) induces cell proliferation together with oxidative stress. The present study investigated the effects of salvianolic acid A (Sal A) and B (Sal B) on the PDGF-induced signaling cascades in hepatic stellate cells (HSCs). HSC-T6, a rat hepatic stellate cell line, was stimulated with PDGF (10 ng/mL). The inhibitory effects of Sal A and B on oxidative stress-related signaling pathways were assessed in vitro. The protein levels were measured by Western blotting. FACS analysis was applied to detect the thioredoxin (Trx) level. Sal A and B showed different inhibitory abilities on the PDGF-related pathway. Sal A inhibited 70-kDa ribosomal S6 kinase (p70(s6k)) and associated proteins. Sal B attenuated PDGF-induced c-jun-N-terminal kinase (JNK), p38, and PKC- δ phosphorylations. Both Sal A and B diminished the activation of PKD, Trx, heme-oxygenase (HO)-1, and Nrf2. Taken together, our results showed that Sal A and B attenuated PDGF-induced ROS formation in HSCs, possibly through different signaling pathways.
Assuntos
Benzofuranos/farmacologia , Ácidos Cafeicos/farmacologia , Lactatos/farmacologia , Cirrose Hepática/prevenção & controle , Fator de Crescimento Derivado de Plaquetas/antagonistas & inibidores , Espécies Reativas de Oxigênio/metabolismo , Salvia miltiorrhiza/química , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Fígado/metabolismo , Cirrose Hepática/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Fosforilação , Raízes de Plantas/química , Fator de Crescimento Derivado de Plaquetas/farmacologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosRESUMO
Triptolide (C38H42O6N2, TP, a diterpene triepoxide derived from Tripterygium wilfordii Hook F.), is a potent immunosuppresive and antiinflammatory agent. The present study investigated whether TP exerted antihepatofibrotic effects in vitro and in vivo. A cell line of rat hepatic stellate cells (HSC-T6) was stimulated with tumor necrosis factor-α (TNF-α) or transforming growth factor (TGF)-ß1. The inhibitory effects of TP on the nuclear factor-κB (NFκB) signaling cascade and fibrosis markers, including α-smooth muscle actin (α-SMA) and collagen, were assessed. An in vivo therapeutic study was conducted in dimethylnitrosamine (DMN)-treated rats. The rats were randomly assigned to one of three groups: control rats, DMN rats receiving vehicle only and DMN rats receiving TP (20 µg/kg). Treatment was given by gavage twice daily for 3 weeks starting 1 week after the start of DMN administration. TP (5-100 nM) concentration-dependently inhibited the NFκB transcriptional activity induced by TNF-α, lipopolysaccharide and phorbol 12-myristate 13-acetate in HSC-T6 cells. In addition, TP also suppressed TNF-α and TGF-ß1-induced collagen deposition and α-SMA secretion in HSC-T6 cells. In vivo, TP treatment significantly reduced hepatic fibrosis scores, collagen contents, IL-6 and TNF-α levels, and the number of α-SMA and NFκB-positive cells in DMN rats. The results showed that TP exerted antifibrotic effects in both HSC-T6 cells and DMN rats.
Assuntos
Diterpenos/farmacologia , Células Estreladas do Fígado/efeitos dos fármacos , Cirrose Hepática Experimental/tratamento farmacológico , Fenantrenos/farmacologia , Actinas/metabolismo , Animais , Linhagem Celular , Colágeno/metabolismo , Dimetilnitrosamina/efeitos adversos , Compostos de Epóxi/farmacologia , Interleucina-6/metabolismo , Fígado/efeitos dos fármacos , Fígado/patologia , Cirrose Hepática Experimental/patologia , Masculino , NF-kappa B/metabolismo , Ratos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta1/farmacologia , Tripterygium/química , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
Increasing NO bioavailability improves hepatic endothelial dysfunction, which ameliorates intrahepatic resistance and portal hypertension. Acute administration of sildenafil increases hepatic production of NO with a reduction in hepatic sinusoid resistance in cirrhotic patients and enhances the vasorelaxation response to NO in cirrhotic rat livers. However, the mechanisms were still unclear. Therefore, our present study aims to evaluate the effects and mechanisms of administration of sildenafil for 1 week on the hepatic microcirculation of cirrhotic rats. Cirrhosis was induced by bile duct ligation with sham-operated rats serving as normal controls. Intrahepatic resistance was evaluated by in situ liver perfusion. Expression of phospho-eNOS (endothelial NO synthase), iNOS (inducible NO synthase), phospho-Akt, PDE-5 (phosphodiesterase-5) and sGC (soluble guanylate cyclase) were determined by Western blot analysis. Biosynthesis of BH4 (tetrahydrobiopterin) and GTPCH-I (GTP cyclohydrolase I) activity were examined by HPLC. Intravital microscopy was used to observe the direct change in hepatic microcirculation. In cirrhotic rat livers, sildenafil treatment increased hepatic sinusoid volumetric flow, NO bioavailability, BH4, GTPCH-I activity, and the protein expression of phospho-Akt, phospho-eNOS and sGC. These events were associated with reduced protein expression of PDE-5, portal perfusion pressure and portal vein pressure. In contrast, sham rats did not produce any significant change in these measurements. In conclusion, sildenafil treatment improves endothelial dysfunction by augmenting NO bioavailability in the hepatic microcirculation.
Assuntos
Circulação Hepática/efeitos dos fármacos , Cirrose Hepática Biliar/fisiopatologia , Inibidores de Fosfodiesterase/farmacologia , Piperazinas/farmacologia , Sulfonas/farmacologia , Resistência Vascular/efeitos dos fármacos , Animais , Disponibilidade Biológica , Esquema de Medicação , Avaliação Pré-Clínica de Medicamentos/métodos , Hemodinâmica/efeitos dos fármacos , Hipertensão Portal/tratamento farmacológico , Hipertensão Portal/etiologia , Hipertensão Portal/fisiopatologia , Circulação Hepática/fisiologia , Cirrose Hepática Biliar/complicações , Cirrose Hepática Biliar/tratamento farmacológico , Masculino , Óxido Nítrico/fisiologia , Inibidores de Fosfodiesterase/uso terapêutico , Piperazinas/uso terapêutico , Pressão na Veia Porta/efeitos dos fármacos , Purinas/farmacologia , Purinas/uso terapêutico , Ratos , Ratos Sprague-Dawley , Citrato de Sildenafila , Sulfonas/uso terapêutico , Resistência Vascular/fisiologiaRESUMO
Activation of hepatic stellate cells (HSCs) plays a crucial role in liver fibrogenesis. armepavine (Arm, C19H23O3N), an active compound from Nelumbo nucifera, has been shown to exert immunosuppressive effects on T lymphocytes and on lupus nephritic mice. The aim of this study was to investigate whether Arm could exert anti-hepatic fibrogenic effects in vitro and in vivo. A cell line of rat HSCs (HSC-T6) was stimulated with tumor necrosis factor-alpha (TNF-alpha) or lipopolysaccharide (LPS) to evaluate the inhibitory effects of Arm. An in vivo therapeutic study was conducted in bile duct-ligated (BDL) rats. BDL rats were given Arm (3 or 10 mg/kg) by gavage twice daily for 3 weeks starting from the onset of BDL. Liver sections were taken for fibrosis scoring, immuno-fluorescence staining and quantitative real-time mRNA measurements. In vitro, Arm (1-10 microM) concentration-dependently attenuated TNF-alpha- and LPS-stimulated alpha-SMA protein expression and AP-1 activation by HSC-T6 cells without adverse cytotoxicity. Arm also suppressed TNF-alpha-induced collagen collagen deposition, NFkappaB activation and MAPK (p38, ERK1/2, and JNK) phosphorylations. In vivo, Arm treatment significantly reduced plasma AST and ALT levels, hepatic alpha-SMA expression and collagen contents, and fibrosis scores of BDL rats as compared with vehicle treatment. Moreover, Arm attenuated the mRNA expression levels of col 1alpha2, TGF-beta1, TIMP-1, ICAM-1, iNOS, and IL-6 genes, but up-regulated metallothionein genes. Our study results showed that Arm exerted both in vitro and in vivo antifibrotic effects in rats, possibly through anti-NF-kappaB activation pathways.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Benzilisoquinolinas/uso terapêutico , Células Estreladas do Fígado/efeitos dos fármacos , Imunossupressores/uso terapêutico , Cirrose Hepática Biliar/tratamento farmacológico , Cirrose Hepática Experimental/tratamento farmacológico , Fitoterapia , Actinas/biossíntese , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Anti-Inflamatórios não Esteroides/toxicidade , Benzilisoquinolinas/farmacologia , Benzilisoquinolinas/toxicidade , Linhagem Celular/efeitos dos fármacos , Linhagem Celular/ultraestrutura , Colágeno/biossíntese , Avaliação Pré-Clínica de Medicamentos , Regulação da Expressão Gênica/efeitos dos fármacos , Células Estreladas do Fígado/ultraestrutura , Imunossupressores/farmacologia , Imunossupressores/toxicidade , Lipopolissacarídeos/farmacologia , Cirrose Hepática Biliar/sangue , Cirrose Hepática Biliar/genética , Cirrose Hepática Biliar/patologia , Cirrose Hepática Experimental/sangue , Cirrose Hepática Experimental/genética , Cirrose Hepática Experimental/patologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , NF-kappa B/metabolismo , Nelumbo/química , Fosforilação , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
Acute liver failure (ALF), an often fatal condition characterized by massive hepatocyte necrosis, is frequently caused by drug poisoning, particularly with acetaminophen (N-acetyl-p-aminophenol/APAP). Hepatocyte necrosis is consecutive to glutathione (GSH) depletion and mitochondrial damage caused by reactive oxygen species (ROS) overproduction. Magnolol, one major phenolic constituent of Magnolia officinalis, have been known to exhibit potent antioxidative activity. In this study, the anti-hepatotoxic activity of magnolol on APAP-induced toxicity in the Sprague-Dawley rat liver was examined. After evaluating the changes of several biochemical parameters in serum, the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH) were elevated by APAP (500 mg/kg) intraperitoneal administration (8 and 24 h) and reduced by treatment with magnolol (0.5 h after APAP administration; 0.01, 0.1, and 1 mug/kg). Histological changes around the hepatic central vein, lipid peroxidation (thiobarbituric acid-reactive substance/TBARS), and GSH depletion in liver tissue induced by APAP were also recovered by magnolol treatment. The data show that oxidative stress followed by lipid peroxidation may play a very important role in the pathogenesis of APAP-induced hepatic injury; treatment with lipid-soluble antioxidant, magnolol, exerts anti-hepatotoxic activity. Our study points out the potential interest of magnolol in the treatment of toxic ALF.
Assuntos
Acetaminofen/toxicidade , Antioxidantes/uso terapêutico , Compostos de Bifenilo/uso terapêutico , Lignanas/uso terapêutico , Falência Hepática Aguda/prevenção & controle , Fígado/efeitos dos fármacos , Animais , Antioxidantes/administração & dosagem , Antioxidantes/isolamento & purificação , Compostos de Bifenilo/administração & dosagem , Compostos de Bifenilo/isolamento & purificação , Glutationa/metabolismo , Lignanas/administração & dosagem , Lignanas/isolamento & purificação , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Falência Hepática Aguda/induzido quimicamente , Falência Hepática Aguda/patologia , Magnolia/química , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND AND AIMS: In chronic liver injury, hepatic stellate cells (HSCs) acquire an activated phenotype, migrate to the injured region in response to chemotactic factors, and produce extracellular matrix proteins including collagen. In this study, we investigated the effects of rhubarb (Rheum palmatum L.) on transforming growth factor (TGF)-beta1-induced expressions of alpha-smooth-muscle actin (SMA) and collagen, and the migration of HSCs. METHODS: HSC-T6, a cell line of rat HSCs, was used in the in vitro experiments. An enzyme-linked immunosorbent assay and Sircol red assay were used to detect the expressions of alpha-SMA and collagen, respectively. HSC-T6 migration was assayed with a transwell apparatus. Phosphorylations of Smad2/3 and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-jun N-terminal kinase (JNK), were analyzed with Western blotting. Matrix metalloproteinase (MMP)-2 activity was examined by gelatin zymography. RESULTS: The results revealed that a rhubarb extract concentration-dependently attenuated TGF-beta1-induced alpha-SMA and collagen expressions and migration of HSCs. The inhibitory effect of rhubarb was associated with (i) down-regulation of the phosphorylation of Smad2/3 and JNK, and (ii) attenuation of MMP-2 activity. Within the working concentrations used, the rhubarb extract did not affect cell viability of HSCs. CONCLUSION: The results suggest that rhubarb attenuated TGF-beta1-mediated migration of HSCs possibly by interfering with Smad2/3 phosphorylation, the MAPK pathway, and MMP-2 activity.
Assuntos
Quimiotaxia/efeitos dos fármacos , Células Estreladas do Fígado/efeitos dos fármacos , Extratos Vegetais/farmacologia , Rheum , Actinas/metabolismo , Animais , Linhagem Celular , Colágeno/metabolismo , Relação Dose-Resposta a Droga , Células Estreladas do Fígado/enzimologia , Células Estreladas do Fígado/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Metaloproteinase 2 da Matriz/metabolismo , Fosforilação , Ratos , Proteína Smad2/metabolismo , Proteína Smad3/metabolismo , Fator de Crescimento Transformador beta1/metabolismoRESUMO
Suppression of activation or fibrogenesis and induction of apoptosis, in hepatic stellate cells (HSCs) have been proposed as therapeutic strategies against liver fibrosis. Curcumin, an active compound isolated from yellow curry pigment of turmeric (Curcuma longa Linn), has been demonstrated to be an effective anti-inflammatory and antioxidant compound. In this study, we investigated the in vitro antifibrogenic effects of curcumin on HSCs at the concentration range of (1-40 microM). A cell line of rat HSCs (HSC-T6) was stimulated with transforming growth factor-beta1 (TGF-beta1). The inhibitory effects of curcumin (1.25 approximately 10 microM) on fibrosis-related markers including alpha-smooth muscle actin (alpha-SMA) and collagen were assessed. In addition, the induction effects of curcumin (20 approximately 40 microM) on apoptosis in HSC-T6 cells were also assessed by Hoechst and propidium iodide stains. Curcumin (1.25 approximately 10 microM) concentration-dependently suppressed TGF-beta1-induced alpha-SMA expression and collagen deposition in HSC-T6 cells, without cytotoxicity. Whereas, higher concentrations of curcumin (20 approximately 40 microM) induced cell apoptosis and cytochrome c release in HSC-T6 cells. Our results suggest that curcumin exerted antifibrotic effects, possibly through two different mechanisms depending on its concentrations. At lower concentrations (1.25 approximately 10 microM), curcumin exerted antifibrogenic effects, whereas at higher concentrations (20 approximately 40 microM), curcumin exerted induction of apoptosis in HSCs.
Assuntos
Apoptose , Curcumina/farmacologia , Células Estreladas do Fígado/efeitos dos fármacos , Actinas/metabolismo , Animais , Linhagem Celular , Colágeno/metabolismo , Citocromos c/metabolismo , Células Estreladas do Fígado/metabolismo , Cirrose Hepática/prevenção & controle , Ratos , Fator de Crescimento Transformador beta1/farmacologiaRESUMO
Liver cirrhosis is characterized by increased IHR (intrahepatic resistance) and lipid peroxidation, and decreased antioxidative defence. The present study investigates the effects of administration for 1 month of the antioxidant UDCA (ursodeoxycholic acid) in BDL (bile-duct-ligated) cirrhotic rats. Splanchnic haemodynamics, IHR, hepatic levels of TBARS (thiobarbituric acid-reacting substances), GSH (glutathione), SOD (superoxide dismutase) activity, nitrite, PIIINP (N-terminal propeptide of type III procollagen) and collagen deposition, histological examination of liver, mRNA expression of PIIIP-alpha1 (type III procollagen) and TGF-beta1 (transforming growth factor-beta1), protein expression of TXS (thromboxane synthase) and iNOS (inducible NO synthase), and TXA2 (thromboxane A2) production in liver perfusates were measured. The results showed that portal pressure and IHR, hepatic levels of PIIINP, hepatic collagen deposition, mRNA expression of PIIIP-alpha1 and TGF-beta1, protein expression of iNOS and TXS, and production of TXA2 in liver perfusates were significantly decreased in UDCA-treated BDL rats. The increased levels of hepatic GSH and SOD activity and decreased levels of TBARS and nitrite were also observed in UDCA-treated BDL rats. In UDCA-treated BDL rats, the reduction in portal pressure resulted from a decrease in IHR, which mostly acted through the suppression of hepatic TXA2 production and lipid peroxidation, and an increase in antioxidative defence, leading to the prevention of hepatic fibrosis.
Assuntos
Antioxidantes/uso terapêutico , Hipertensão Portal/tratamento farmacológico , Cirrose Hepática Biliar/tratamento farmacológico , Ácido Ursodesoxicólico/uso terapêutico , Animais , Antioxidantes/administração & dosagem , Antioxidantes/metabolismo , Colágeno Tipo III/biossíntese , Colágeno Tipo III/genética , Esquema de Medicação , Avaliação Pré-Clínica de Medicamentos , Regulação da Expressão Gênica/efeitos dos fármacos , Hemodinâmica/efeitos dos fármacos , Hipertensão Portal/etiologia , Hipertensão Portal/fisiopatologia , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Fígado/fisiopatologia , Circulação Hepática/efeitos dos fármacos , Cirrose Hepática Biliar/complicações , Cirrose Hepática Biliar/patologia , Cirrose Hepática Biliar/fisiopatologia , Masculino , Fragmentos de Peptídeos/biossíntese , Fragmentos de Peptídeos/genética , Pressão na Veia Porta/efeitos dos fármacos , Pró-Colágeno/biossíntese , Pró-Colágeno/genética , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Ácido Ursodesoxicólico/administração & dosagem , Resistência Vascular/efeitos dos fármacosRESUMO
BACKGROUND: In 1995, Taiwan has launched a national health-care system (the National Health Insurance Program, NHI) covering the use of both Western medicine (WM) and Chinese medicine (CM). This population-based study was conducted to understand the role of CM in this dual medical system by determining the utilization patterns of CM and WM and to analyze the demographic characteristics and primary indications influencing the choice of the medical services for the development of strategies to enhance the appropriate use and reduce unnecessary use of CM. METHODS: This study used the NHI sample files from 1997 to 2003 consisting of comprehensive utilization and enrolment information for a random sample of 200,432 NHI beneficiaries of the total enrolees from 1995 to 2000. A total of 136,720 subjects with valid and complete enrolment and utilization data were included in this study. The logistic regression method was employed to estimate the odds ratios (ORs) for utilization of CM and WM. The usage, frequency of services, and primary indications for CM and WM were evaluated. A significance level of alpha = 0.05 was selected. RESULTS: Compared with WM, the odds of CM increased from 1997 to 2003. The odds of using CM (OR = 1.48; 95% CI: 1.45-1.50; p < 0.001) and WM (OR = 1.74; 95% CI: 1.72-1.77; p < 0.001) were higher in females and that of CM increased with age to a peak in the 45-54-year-group (OR = 1.75; 95% CI: 1.68-1.82; p < 0.001) and WM (OR = 1.09; 95% CI: 1.05-1.13; p < 0.001) in the elderly subjects (> or = 65 years). The odds of CM and WM were similar in all income groups. However, those of CM were higher in Central (OR = 1.65; 95% CI: 1.56-1.74; p < 0.001) and Southern Taiwan (OR = 1.18; 95% CI: 1.12-1.25; p < 0.001) and lower in the remote areas (OR = 0.57; 95% CI: 0.52-0.63; p < 0.001). Most of the patients had one ambulatory visit of both medical services annually. However, the utilization of WM predominated over CM. Over 90% of CM service was provided by clinics, whereas over 60% of WM service by hospitals. Diseases of the respiratory system was the most frequent primary indication in CM and WM. Herbal medication was the most commonly used form of CM (68.4-72.7%). CONCLUSION: In recent years, there is an increasing trend in the utilization of CM in Taiwan. This increasing trend may be due to the covering of CM in the national health insurance system.
Assuntos
Medicina Tradicional Chinesa/estatística & dados numéricos , Programas Nacionais de Saúde , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Estudos de Coortes , Feminino , Serviços de Saúde/estatística & dados numéricos , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Classe Social , Taiwan , OcidenteRESUMO
In liver injury, hepatic stellate cells (HSCs) acquire an activated phenotype, migrate to the injured region in response to chemotactic factors and produce extracellular matrix (ECM) proteins including alpha-smooth muscle actin (alpha-SMA) and collagen in order to repair the damage. HSC-T6, a cell line of rat HSCs, was used in in vitro experiments. TGF-beta1 was used as a chemoattractant. The expression of alpha-SMA was used as a marker of activated hepatic stellate cells and cell migration was assayed with the Transwell method to investigate the active principles of the roots of Rheum palmatum L. (Dahuang), a well-known traditional Chinese herb used for treating liver diseases. Under cell activation and chemotaxis-directed fractionation and purification, four anthraquinones, rhein ( 1), emodin ( 2), chrysophanol ( 3) and physcion ( 4), and four phenylbutanoids, lindleyin ( 5), isolindleyin ( 7), 4-(4'-hydroxyphenyl)-2-butanone 4'- O-beta- D-glucopyranoside ( 8), and 4-(4'-hydroxyphenyl)-2-butanone ( 9), and a stilbene, 3,5,4'-trihydroxystilbene 4'- O-beta- D-glucopyranoside 6'- O-gallate ( 6) were isolated from the active fractions. Among them, compounds 1 and 2 inhibited alpha-SMA expression. However, compounds 3, 4, 6 and 8 attenuated chemotactic migration, but not alpha-SMA expression.
Assuntos
Actinas/metabolismo , Quimiotaxia/efeitos dos fármacos , Hidrocarbonetos Aromáticos/farmacologia , Rheum/química , Fator de Crescimento Transformador beta1/metabolismo , Animais , Linhagem Celular , Medicamentos de Ervas Chinesas/uso terapêutico , Matriz Extracelular/metabolismo , Hidrocarbonetos Aromáticos/isolamento & purificação , Hidrocarbonetos Aromáticos/uso terapêutico , Fígado/citologia , Cirrose Hepática/tratamento farmacológico , Fenóis/isolamento & purificação , Fenóis/farmacologia , Fenóis/uso terapêutico , Fitoterapia , Raízes de Plantas/química , Ratos , Ratos Sprague-DawleyRESUMO
Liver fibrosis has been characterized as chronic inflammatory processes involving multiple molecular pathogenetic pathways. This therapeutic study investigated whether a combination regimen of Salvia miltiorrhiza (S), Ligusticum chuanxiong (L) and Glycyrrhiza glabra (G) exerted in vivo antifibrotic effects on rats with hepatic fibrosis. Fibrosis was induced in rats by dimethylnitrosamine (DMN) administration for 4 weeks. Fibrotic rats were randomly assigned to one of the three groups: control, SLG (50 mg/kg) or silymarin (50 mg/kg), each given by gavage twice daily for 3 weeks starting 1 week after DMN injection. The results showed that fibrosis scores of livers from DMN-treated rats with SLG (1.13 +/- 0.13) were significantly reduced in comparison with DMN-treated rats receiving vehicle (1.63 +/- 0.18). Moreover, the hepatic collagen content of DMN rats was significantly reduced by either SLG or silymarin treatment. The double immunohistochemical staining results also showed that alpha-SMA positive cells with NF kappa B nuclear translocation were decreased in the fibrotic livers by SLG and silymarin treatments. The mRNA expression levels of TGF-beta1, alpha-SMA, collagen1 alpha 2, iNOS and ICAM-1 genes were attenuated by SLG and silymarin treatment. The results showed that SLG exerted antifibrotic effects in rats with DMN-induced hepatic fibrosis.