RESUMO
Panax ginseng has a wide range of activities including a neuroprotective effect, skin protective effects, enhanced DNA repairing, anti-diabetic activity, and protective effects against vascular inflammation. In the present study, we sought to discover the inhibitory effects of a mixture of natural products containing Panax ginseng, Ziziphus jujube, Rubi fructus, Artemisiae asiaticae and Scutellaria baicalensis (PZRAS) on osteoclastogenesis and bone remodeling, as neither the effects of a mixture containing Panax ginseng extract, nor its molecular mechanism on bone inflammation, have been clarified yet. PZRAS upregulated the levels of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GSH-R) and glutathione peroxidase (GSH-Px) and reduced malondialdehyde (MDA) in LPS-treated RAW264.7 cells. Moreover, treatment with PZRAS decreased the production of IL-1ß and TNF-α. PZRAS also inhibited osteoclast differentiation through inhibiting osteoclastspecific genes like MMP-2, 9, cathepsin K, and TRAP in RANKL-treated RAW264.7 cells. Additionally, PZRAS has inhibitory functions on the RANKL-stimulated activation of ERK and JNK, which lead to a decrease in the expression of NFATc1 and c-Fos. In an in vivo study, bone resorption induced by LPS was recovered by treatment with PZRAS in bone volume per tissue volume (BV/TV) compared to control. Furthermore, the ratio of eroded bone surface of femurs was significantly increased in LPStreated mice compared to vehicle group, but this ratio was significantly reversed in PZRAS-treated mice. These results suggest that PZRAS could prevent or treat disorders with abnormal bone loss.
Assuntos
Reabsorção Óssea/prevenção & controle , Inflamação/prevenção & controle , Osteogênese/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Catepsina K/genética , Catepsina K/metabolismo , Diferenciação Celular/efeitos dos fármacos , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Lipopolissacarídeos/toxicidade , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Fatores de Transcrição NFATC/genética , Fatores de Transcrição NFATC/metabolismo , Proteínas Proto-Oncogênicas c-fos/genética , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ligante RANK/genética , Ligante RANK/metabolismo , Células RAW 264.7 , Fosfatase Ácida Resistente a Tartarato/genética , Fosfatase Ácida Resistente a Tartarato/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: Grapes are among the most widely consumed plants and are used as a folk medicine. Vitis species have been traditionally used as anti-inflammatory, analgesic, and memory-enhancing agents, but, their biological activities of discarded grape leaves are not completely understood. PURPOSE: We investigated the effects of alcoholic aqueous leaf extract of Vitis labruscana (LEVL) in a mouse model of memory impairment and tried to ascertain its mechanism. We also evaluated its effects in SH-SY5Y cells. METHODS: LEVL (50, 100, and 150â¯mg/kg) was administered to ICR mice once daily for 7 days. Memory impairment was induced with intraperitoneal scopolamine injections (1â¯mg/kg) and measured with the Y-maze test and a passive avoidance task. LEVL-induced signaling was evaluated in SH-SY5Y cells and mouse hippocampi. RESULTS: We first identified quercetin-3-O-glucuronide as LEVL's major component. We then showed that LEVL promoted phosphorylation of Akt, extracellular regulated kinase (ERK), and cyclic AMP response element binding protein (CREB) and proliferation of SH-SY5Y cells. Oral LEVL administration (100â¯mg/kg) for 7 days significantly reversed scopolamine-induced reductions of spontaneous alternation in the Y-maze test and scopolamine-induced shortening of latency times in the passive avoidance task's retention trial. Consistent with the cell experiment results, LEVL restored scopolamine-decreased phosphorylation of Akt, ERK, and CREB and scopolamine-reduced expression of brain-derived neuroprotective factor expression in mouse hippocampi. CONCLUSION: Our results suggest that LEVL promotes phosphorylation of Akt, ERK, and CREB in the hippocampus and ameliorates scopolamine-induced memory impairment in mice.
Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Transtornos da Memória/tratamento farmacológico , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Vitis/química , Animais , Linhagem Celular , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Hipocampo/efeitos dos fármacos , Humanos , Espectroscopia de Ressonância Magnética , Masculino , Memória/efeitos dos fármacos , Transtornos da Memória/induzido quimicamente , Camundongos Endogâmicos ICR , Fosforilação , Extratos Vegetais/química , Folhas de Planta/química , Escopolamina/efeitos adversos , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND: Kudzu (Pueraria thunbergiana) root has long been used in Traditional Chinese Medicine. However, the vine of the kudzu plant has been considered waste material. This study aimed to investigate the hepatoprotective properties of the kudzu vine. METHODS: We created 0 %, 30 %, 70 %, and 95 % ethanolic kudzu vine extracts. The isoflavone contents of kudzu vine extract were quantified by high-performance liquid chromatography. Tertiary-butylhydroperoxide (t-BHP) was added to human liver-derived HepG2 cells, and the production of reactive oxygen species was measured in the presence and absence of kudzu vine extract. Antioxidant activity was evaluated in all kudzu vine extracts using a hydroxyradical scavenging assay. Thirty-five male Sprague-Dawley rats were divided into seven groups (n = 5); two groups were not given any extract or drug, one group was treated with 50 mg/kg silymarin orally for 5 days, and the remaining four groups were respectively treated with 100 mg/kg of 0%, 30%, 70%, or 95% ethanolic extract of kudzu vine orally once daily for 5 days. On day 5 the treatment groups and one untreated group were fed 0.75 ml/kg carbon tetrachloride (CCl4) to induce liver damage. Blood and liver tissue samples were collected 24 h after CCl4 administration for measurement of plasma alanine aminotransferase and aspartate aminotransferase, and concentration of malondialdehyde and glutathione in liver tissue. RESULTS: Puerarin was the most abundant isoflavone in kudzu vine extract. Kudzu vine extract significantly reduced the cytotoxicity and production of reactive oxygen species induced by t-BHP in a dose-dependent manner. Treatment with 0 % and 30 % ethanolic extracts of kudzu vine significantly lowered the plasma levels of alanine aminotransferase and aspartate aminotransferase in a CCl4-induced hepatotoxicity rat model (P < 0.05). Glutathione was significantly elevated in the 30 % ethanolic extract-treated group (P < 0.05), while the malondialdehyde level in liver tissue was significantly decreased in the 0 % and 30 % ethanolic extract-treated groups (P < 0.05). CONCLUSIONS: The kudzu vine is potentially highly beneficial in treating liver damage, as it scavenges reactive free radicals and boosts the endogenous antioxidant system.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Fígado/efeitos dos fármacos , Substâncias Protetoras/uso terapêutico , Pueraria/química , Animais , Antioxidantes/metabolismo , Antioxidantes/uso terapêutico , Tetracloreto de Carbono , Medicamentos de Ervas Chinesas , Sequestradores de Radicais Livres/metabolismo , Células Hep G2 , Humanos , Isoflavonas/análise , Isoflavonas/uso terapêutico , Masculino , Ratos Sprague-Dawley , terc-Butil HidroperóxidoRESUMO
Vitis labrusca is a grapevine that has antioxidant, neuroprotective, hepatoprotective, and anticarcinogenic activity. However, the antithrombotic effect of Vitis labrusca leaves on platelets is yet to be ascertained. We investigated the inhibitory effect of V. labrusca leaf extract (VLE) on platelet aggregation in vitro and ex vivo. The thromboxane B2 (TXB2) and serotonin concentrations were measured by ELISA. The flavonoids content was measured by ultraperformance liquid chromatography (UPLC). The antithrombotic activity of VLE was evaluated using various agonists in vitro. VLE strongly inhibited adenosine diphosphate (ADP)-induced platelet aggregation. In rats, VLE treatment (100âmg/kg) reduced ADP-stimulated platelet aggregation, without affecting tail bleeding and coagulation time. Moreover, VLE significantly suppressed TXB2 and serotonin secretion. UPLC analysis indicated that VLE contains quercetin, isorhamnetin, and rutin. Our results indicate that VLE possesses antiplatelet activity via the suppression of TXB2 and serotonin, without affecting bleeding. Further, we identified the flavonoids present in VLE. Thus, VLE may be a potential agent for the prevention of cardiovascular diseases.
Assuntos
Plaquetas/efeitos dos fármacos , Flavonoides/farmacologia , Extratos Vegetais/farmacologia , Inibidores da Agregação Plaquetária/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Vitis/química , Difosfato de Adenosina/farmacologia , Animais , Relação Dose-Resposta a Droga , Flavonoides/química , Flavonoides/isolamento & purificação , Masculino , Extratos Vegetais/química , Folhas de Planta/química , Inibidores da Agregação Plaquetária/química , Inibidores da Agregação Plaquetária/isolamento & purificação , Ratos , Ratos Sprague-Dawley , Serotonina/metabolismo , Tromboxano B2/antagonistas & inibidores , Tromboxano B2/metabolismoRESUMO
The seeds of Phaseolus calcaratus Roxburgh (PHCR) are common legumes that comprise part of the daily diet in Chinese and Korean culture. Recent findings highlight anti-inflammatory and anti-septic potentials of catechin-7-O-ß-D-glucopyranoside (CGP) isolated from PHCR seeds. We investigated the intestinal anti-inflammatory activity and associated mechanisms of CGP using a rat model of trinitrobenzenesulfonic acid (TNBS)-induced colitis. Oral treatment with CGP (10mg/kg body weight) suppressed body weight loss and intestinal inflammatory damages in TNBS-induced colitic rats. This treatment reduced myeloperoxidase activity and malondialdehyde level, but increased glutathione level in the TNBS colitic rats. CGP treatment also inhibited the TNBS-mediated increases in nitric oxide synthase, cyclooxygenase-2, interleukin-1ß, tumor necrosis factor-α, intercellular adhesion molecule-1, and monocyte chemotactic protein-1 proteins or mRNA levels. This inhibition was accompanied by the increased mRNA levels of mucins MUC2 and MUC3. The CGP treatment prevented phosphorylation of p38 mitogen-activated protein kinase, IκB-α, and DNA-nuclear factor-κB binding, all of which were increased in the inflamed colons of TNBS-treated rats. Furthermore, oral administration with a crude PHCR butanol extract (100mg/kg body weight) which contains 1.5% of CGP showed intestinal anti-inflammatory potentials similar to that of CGP. Collectively, our current findings suggest that CGP or CGP-containing PHCR seeds may have favorable effects on intestinal inflammatory diseases.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Catequina/análogos & derivados , Doença de Crohn/induzido quimicamente , Doença de Crohn/tratamento farmacológico , Glucosídeos/uso terapêutico , Phaseolus/química , Animais , Peso Corporal/efeitos dos fármacos , Catequina/isolamento & purificação , Catequina/uso terapêutico , Regulação da Expressão Gênica/efeitos dos fármacos , Glucosídeos/isolamento & purificação , Glutationa/metabolismo , Masculino , Malondialdeído/metabolismo , Mucinas/biossíntese , Peroxidase/metabolismo , Fosforilação , RNA Mensageiro/biossíntese , Ratos , Ratos Sprague-Dawley , Sementes/química , Ácido TrinitrobenzenossulfônicoRESUMO
The seeds of Raphanus sativus L. (RSL) have long been used as anti-inflammatory traditional medicine. However, scientific bases for the purported potential of the medicine and the associated mechanisms were barely defined. This study investigated the effects of RSL seeds on lipopolysaccharide (LPS)-stimulated inflammatory responses in vitro and in vivo. Treatment with 100 µg/ml ethyl acetate fraction (REF), which was isolated from water extract of the seeds, significantly inhibited LPS-stimulated production of nitric oxide (P < 0.05), interleukin-6 (P < 0.001), and tumor necrosis factor (TNF)-α (P < 0.001) in RAW264.7 cells. Oral supplementation with 30 mg/kg REF protected mice by 90% against LPS-induced septic death and prevented the increases of serum TNF-α and interferon-γ levels in LPS-injected mice. When REF was divided into four sub-fractions (REF-F1-F4), REF-F3 showed the greatest activity to suppress LPS-stimulated production of inflammatory mediators. We subsequently isolated an active fraction from the REF-F3 and identified sinapic acid as the main constituent. The addition of 50 µg/ml active fraction markedly inhibited LPS-stimulated production of inflammatory mediators by suppressing p38 MAPK and nuclear factor-κB activation. Furthermore, supplementation with the active fraction (10 mg/kg) improved the survival rate of LPS-injected mice by 80% of the untreated control. Additional experiments revealed that sinapic acid was the active component responsible for the anti-inflammatory potential of RSL seeds. Collectively, our current results suggest that both RSL seeds and sinapic acid may be attractive materials for treating inflammatory disorders caused by endotoxins.
Assuntos
Anti-Inflamatórios/uso terapêutico , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , NF-kappa B/metabolismo , Extratos Vegetais/uso terapêutico , Raphanus/química , Sepse/tratamento farmacológico , Animais , Anti-Inflamatórios/administração & dosagem , Antioxidantes/metabolismo , Linhagem Celular , Citocinas/sangue , Feminino , Lipopolissacarídeos/toxicidade , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Camundongos Endogâmicos ICR , Óxido Nítrico/metabolismo , Extratos Vegetais/administração & dosagem , Sementes/química , Sepse/induzido quimicamente , Sepse/enzimologia , Sepse/imunologiaRESUMO
Endochondral bone formation occurs when mesenchymal cells condense to differentiate into chondrocytes, the primary cell types of cartilage. The aim of the present study was to identify novel factors regulating chondrogenesis. We investigated whether kaempferol induces chondrogenic differentiation in clonal mouse chondrogenic ATDC5 cells. Kaempferol treatment stimulated the accumulation of cartilage nodules in a dose-dependent manner. Kaempferol-treated ATDC5 cells stained more intensely with alcian blue staining than control cells, suggesting greater synthesis of matrix proteoglycans in the kaempferol-treated cells. Similarly, kaempferol induced greater activation of alkaline phosphatase activity than control cells, and it enhanced the expression of chondrogenic marker genes, such as collagen type I, collagen type X, OCN, Runx2, and Sox9. Kaempferol induced an acute activation of extracellular signal-regulated kinase (ERK) but not c-jun N-terminal kinase or p38 MAP kinase. PD98059, an inhibitor of MAPK/ERK, decreased in stained cells treated with kaempferol. Furthermore, kaempferol greatly expressed the protein and mRNA levels of BMP-2, suggesting chondrogenesis was stimulated via a BMP-2 pathway. Taken together, our results suggest that kaempferol has chondromodulating effects via an ERK/BMP-2 signaling pathway and could potentially be used as a therapeutic agent for bone growth disorders.
Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Condrogênese/efeitos dos fármacos , Quempferóis/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Animais , Cartilagem/citologia , Cartilagem/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular/efeitos dos fármacos , Condrogênese/genética , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Camundongos , Proteoglicanas/metabolismo , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
OBJECTIVES: The seed of Phaseolus calcaratus Roxburgh (PHCR) has traditionally been used as a herbal medicine, considered to have anti-inflammatory potential. Here we examined the ability of PHCR seed extract to inhibit inflammatory responses of macrophages to bacterial toxin and the mechanism involved. METHODS: In the present study, we prepared four fractions from an ethanol extract of PHCR seed and investigated their effects on the production of nitric oxide and cytokines, and the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. KEY FINDINGS: The fractions inhibited LPS-induced nitric oxide production and cyclooxygenase-2 (COX-2) expression in the cells. The ethyl acetate fraction at 100 µg/ml almost completely suppressed NO production, iNOS and COX-2 expression, and TNF-α and IL-6 secretion in cells stimulated with LPS. The fraction also inhibited phosphorylation of extracellular signal-regulated kinase (ERK) and p38 in LPS-stimulated cells with the attendant suppression of IκBα nuclear translocation and nuclear factor (NF)-κB activation. Furthermore, PHCR seed extracts contained a large number of phenolic compounds having antioxidant potentials against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and hydroxyl radicals. We identified catechin-7-O-ß-D-glucopyranoside as one of the active compounds responsible for the biological activity of PHCR seed extract. CONCLUSIONS: These results suggest for the first time that ethanol extracts from PHCR seed have anti-inflammatory potential on LPS-stimulated macrophages through the down-regulation of ERK/p38- and NF-κB-mediated signalling pathways.
Assuntos
Anti-Inflamatórios/farmacologia , Macrófagos/efeitos dos fármacos , Phaseolus/química , Extratos Vegetais/farmacologia , Animais , Anti-Inflamatórios/isolamento & purificação , Citocinas/metabolismo , Regulação para Baixo/efeitos dos fármacos , MAP Quinases Reguladas por Sinal Extracelular/efeitos dos fármacos , MAP Quinases Reguladas por Sinal Extracelular/genética , Lipopolissacarídeos , Macrófagos/metabolismo , Medicina Tradicional do Leste Asiático , Camundongos , NF-kappa B/efeitos dos fármacos , NF-kappa B/genética , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Sementes , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Jangwonhwan, a boiled extract of 12 medicinal plants/mushroom including Korean red ginseng (Panax ginseng C.A. Meyer), has been prescribed for patients with cognitive dysfunction and are believed to induce brain activity enhancement, provide light sedation, and facilitate sound sleep. AIM OF THE STUDY: The present study was carried out to investigate whether Jangwonhwan has a beneficial effect on the brain of Alzheimer disease. MATERIALS AND METHODS: The transgenic mice of Alzheimer disease, Tg-APPswe/PS1dE9, were fed a modified recipe of Jangwonhwan consisting of a boiled extract of 7 herbs/mushroom (called LMK02-Jangwonhwan) at 400mg/kg/day of dose for 3 months from 4.5 months of age. Immunohistological and ELISA analyses were used to assess the Abeta accumulation and plaque deposition in the brain. Other in vitro and in vivo works were performed to understand the underlying mechanism. RESULTS: LMK02-Jangwonhwan notably reduced Abeta(1-42) and Abeta(1-40) levels, concomitantly with a reduction of plaque deposition, in the brain of Tg-APPswe/PS1dE9 mice. LMK02-Jangwonhwan partially suppressed oxidative stress accumulation, and prevented the down-regulation of phospho-CREB and calbindin typically seen in the hippocampus of AD-like brains. In vitro study with SH-SY5Y neuroblastoma cells showed that LMK02-Jangwonhwan inhibited oxidative stress and Abeta-induced neurotoxicity. CONCLUSION: The present study suggests that LMK02-Jangwonhwan confers a therapeutic potential to ameliorate AD-like pathology in the brain of Tg-APPswe/PS1dE9 mice.