RESUMO
Sarcopenia, loss of muscle mass and function, is mainly observed in elderly people. In this study, we investigated whether fermented rice germ extract (FRGE) has some effects on the mouse gastrocnemius muscle by using behavioral and morphological analyses, Western blotting, and a murine model of immobilization-induced muscle atrophy. Daily oral FRGE administration increased muscle weight and strength. In addition, myofiber size in gastrocnemius muscle of FRGE-treated mice was increased as revealed by morphological quantification. Activation of AMP-activated protein kinase (AMPK) signaling, which inhibits protein synthesis and stimulates protein degradation in gastrocnemius muscle, was significantly attenuated in the FRGE-treated mice compared with control mice. Expression level of forkhead box 3a (FOXO3a) protein was also significantly decreased in the FRGE-treated group. Moreover, the decrease in mean myofiber cross-sectional area in immobilized hindlimb in vehicle-treated mice was inhibited by FRGE treatment in histological analysis. In conclusion, FRGE increased the strength and weight of gastrocnemius muscle and myofiber size, and reduced immobilization-induced muscle atrophy in mice. These findings indicated that FRGE might be beneficial in preventing motor dysfunction in a range of conditions, including sarcopenia.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Músculo Esquelético/enzimologia , Oryza/química , Extratos Vegetais/administração & dosagem , Sarcopenia/tratamento farmacológico , Proteínas Quinases Ativadas por AMP/genética , Animais , Aspergillus oryzae/metabolismo , Fermentação , Proteína Forkhead Box O3/genética , Proteína Forkhead Box O3/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Força Muscular , Músculo Esquelético/anatomia & histologia , Músculo Esquelético/fisiopatologia , Oryza/microbiologia , Fosforilação , Extratos Vegetais/metabolismo , Sarcopenia/enzimologia , Sarcopenia/genética , Sarcopenia/fisiopatologiaRESUMO
BACKGROUND: Intracerebral hemorrhage (ICH) during oral anticoagulation therapy with an oral vitamin K epoxidase reductase such as warfarin is a life-threatening complication. However, whether direct oral anticoagulants (DOACs) are associated with larger hematoma volume and higher mortality rates remains controversial. We evaluated the hematoma volume and pathophysiology of ICH during anticoagulation with warfarin or rivaroxaban, an orally active direct factor Xa inhibitor. METHOD: Mice were orally pretreated with rivaroxaban (10 or 30 mg/kg), warfarin (4 mg/kg), or vehicle. ICH was induced by intrastriatal collagenase-injection. Hematoma volume and neurological deficits 24 h after ICH induction were significantly decreased in the rivaroxaban-pretreated group in comparison with the warfarin-pretreated group. Rivaroxaban did not increase the hematoma volume relative to that observed for vehicle, and improved survival rate 7 days after ICH induction compared with warfarin. RESULT: We evaluated blood-brain barrier (BBB) permeability 6 h after ICH induction using Evans blue spectrophotometry. Evans blue extravasation was significantly reduced in the rivaroxaban group compared with the warfarin group. To investigate the mechanism underlying hematoma expansion and BBB permeability, we focused on thrombin, a clot-derived factor and one of the major contributors to ICH-induced brain injury. To investigate the effects of anticoagulant agents on thrombin-induced injuries, human brain endothelial cells were used in membrane permeability assays. Rivaroxaban, but not warfarin, significantly mitigated the thrombin-induced increase in membrane permeability. CONCLUSION: These findings indicate that rivaroxaban decreases BBB disruption after ICH, and limits early hematoma expansion in these experimental models compared with warfarin. Our study suggests that rivaroxaban has advantages over warfarin with respect to ICH, an important complication during long-term anticoagulation therapy.
Assuntos
Barreira Hematoencefálica/efeitos dos fármacos , Hemorragia Cerebral/tratamento farmacológico , Hemorragia Cerebral/patologia , Inibidores do Fator Xa/uso terapêutico , Hematoma/tratamento farmacológico , Rivaroxabana/uso terapêutico , Animais , Anticoagulantes/uso terapêutico , Barreira Hematoencefálica/fisiopatologia , Células Cultivadas , Hemorragia Cerebral/complicações , Hemorragia Cerebral/mortalidade , Modelos Animais de Doenças , Impedância Elétrica , Células Endoteliais/efeitos dos fármacos , Hematoma/etiologia , Isotiocianatos/farmacocinética , Camundongos , Exame Neurológico , Permeabilidade/efeitos dos fármacos , Resultado do Tratamento , Varfarina/uso terapêuticoRESUMO
Tomato (Solanum lycopersicum) is rich in anthocyanins, which are polyphenolic pigments. This study aimed to analyze and characterize the anthocyanin composition in cultivated blue tomato in Japan. The extracts of peel, seed, and pulp of tomatoes were purified following which anthocyanins and lycopene contents were analyzed using high-performance liquid chromatography and electrospray ionization mass spectrometry. Eleven types of anthocyanins were identified, including delphinidin, petunidin, and malvidin. Further, the antioxidant activity of anthocyanins was evaluated using 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt radical quenching assays and electron spin resonance. "Blue tomato" extracts exert antioxidant activity. Thus, we showed that petunidin was present in the "blue tomato" peel while lycopene was present in the peel and pulp. Additionally, the blue tomato peel extract was found to significantly inhibit H2O2-induced cell death in vitro. This is the first study on cell protective effects of Japanese blue tomato extract and petunidin in murine photoreceptor cells.
Assuntos
Antocianinas/isolamento & purificação , Antioxidantes/isolamento & purificação , Carotenoides/isolamento & purificação , Frutas/química , Solanum lycopersicum/química , Animais , Antocianinas/farmacologia , Antioxidantes/farmacologia , Benzotiazóis/antagonistas & inibidores , Benzotiazóis/metabolismo , Carotenoides/farmacologia , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Peróxido de Hidrogênio/antagonistas & inibidores , Peróxido de Hidrogênio/farmacologia , Licopeno , Camundongos , Células Fotorreceptoras de Vertebrados/citologia , Células Fotorreceptoras de Vertebrados/efeitos dos fármacos , Células Fotorreceptoras de Vertebrados/fisiologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Espectrometria de Massas por Ionização por Electrospray , Ácidos Sulfônicos/antagonistas & inibidores , Ácidos Sulfônicos/metabolismoRESUMO
BACKGROUND: Exposure to ultraviolet A (UVA) irradiation is the major cause of human skin aging. Suppression of UVA irradiation-induced skin fibroblast cell damage protects the skin against aging. An oxidative stress response transcription factor nuclear factor-(erythroid-derived 2)-related factor 2 (Nrf2) has an important role as a cytoprotective system against oxidative stress in the human skin and other organs. Propolis has been commonly used as a traditional medicine since ancient times. The water extract of propolis (WEP) mainly contains caffeoylquinic acids. In our previous study, we reported that WEP and its major constituents protected immortalized human skin fibroblast cells (NB1-RGB) against UVA irradiation-induced cell death. In this study, we examined the mechanism of WEP-mediated skin protection and the possible involvement of Nrf2/antioxidant response element (ARE) pathways. METHODS: Brazilian green propolis was used in the present study (Minas Gerais State, Brazil), Baccharis dracunculifolia is its main source. The Baccharis propolis was extracted with water at 50 °C to yield water extract. The NB1-RGB cell cultures were incubated for 23 h. After replenishing the medium, WEP or its constituents were added to the cell cultures. After 1 h, the cells were exposed to 10 J/cm(2) of UVA light (365 nm UVA light source, CL-1000 L UV Closslinkers, Ultraviolet Products Ltd., Cambridge, UK). Heme oxygenase-1 (HO-1) expression levels in NB1-RGB cells were evaluated using western blotting. Nrf2 nuclear translocation changes in NB1-RGB cells were indicated using immunostaining. RESULTS: We demonstrated that WEP pretreatment up-regulated HO-1 expression level after UVA irradiation at earlier time points than vehicle pretreatment did, and three main constituents of WEP showed similar effects. Furthermore, WEP pretreatment also accelerated Nrf2 nuclear translocation after UVA irradiation. CONCLUSIONS: Our findings indicated that WEP acts as an early inducer of HO-1 and rapid activator of Nrf2 to protect against UVA-induced oxidative stress.
Assuntos
Antioxidantes/farmacologia , Heme Oxigenase-1/biossíntese , Fator 2 Relacionado a NF-E2/biossíntese , Própole , Elementos de Resposta Antioxidante/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Células Cultivadas , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Humanos , Estresse Oxidativo/efeitos da radiação , Extratos Vegetais/farmacologia , Ácido Quínico/análogos & derivados , Ácido Quínico/farmacologia , Pele/efeitos dos fármacos , Pele/metabolismo , Raios Ultravioleta/efeitos adversos , Regulação para Cima/efeitos dos fármacosRESUMO
PURPOSE: Photobiomodulation is the treatment with light in the far-red to near-infrared region of the spectrum and has been reported to have beneficial effects in various animal models of disease, including an age-related macular degeneration (AMD) mouse model. Previous reports have suggested that phagocytosis is reduced by age-related increased oxidative stress in AMD. Therefore, we investigated whether photobiomodulation improves phagocytosis caused by oxidative stress in the human retinal pigment epithelial (ARPE-19) cell line. METHODS: ARPE-19 cells and human primary retinal pigment epithelium (hRPE) cells were incubated and irradiated with near-infrared light (670 nm LED light, 2,500 lx, twice a day, 250 s/per time) for 4 d. Next, hydrogen peroxide (H2O2) and photoreceptor outer segments (POS) labeled using a pH-sensitive fluorescent dye were added to the cell culture, and phagocytosis was evaluated by measuring the fluorescence intensity. Furthermore, cell death was observed by double staining with Hoechst33342 and propidium iodide after photobiomodulation. CM-H2DCFDA, JC-1 dye, and CCK-8 were added to the cell culture to investigate the reactive oxygen species (ROS) production, mitochondrial membrane potential, and cell viability, respectively. We also investigated the expression of phagocytosis-related proteins, such as focal adhesion kinase (FAK) and Mer tyrosine kinase (MerTK). RESULTS: Oxidative stress inhibited phagocytosis, and photobiomodulation increased the oxidative stress-induced hypoactivity of phagocytosis in ARPE-19 cells and hRPE cells. Furthermore, H2O2 and photobiomodulation did not affect cell death in this experimental condition. Photobiomodulation reduced ROS production but did not affect cell viability or mitochondrial membrane potential. The expression of phosphorylated MerTK increased, but phosphorylated FAK was not affected by photobiomodulation. CONCLUSIONS: These findings indicate that near-infrared light photobiomodulation (670 nm) may be a noninvasive, inexpensive, and easy adjunctive therapy to help inhibit the development of ocular diseases induced by the activation of phagocytosis.
Assuntos
Raios Infravermelhos , Fagocitose/efeitos da radiação , Epitélio Pigmentado da Retina/fisiologia , Epitélio Pigmentado da Retina/efeitos da radiação , Animais , Linhagem Celular , Quinase 1 de Adesão Focal/metabolismo , Humanos , Raios Infravermelhos/uso terapêutico , Degeneração Macular/patologia , Degeneração Macular/fisiopatologia , Degeneração Macular/prevenção & controle , Potencial da Membrana Mitocondrial/efeitos da radiação , Camundongos , Estresse Oxidativo/efeitos da radiação , Fototerapia/métodos , Proteínas Proto-Oncogênicas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Receptores Proteína Tirosina Quinases/metabolismo , Epitélio Pigmentado da Retina/citologia , Suínos , c-Mer Tirosina QuinaseRESUMO
Huperzia serrata has been used as a Chinese folk medicine for many years. It contains huperzine A, which has a protective effect against memory deficits in animal models; however, it is unclear if H. serrata extract exerts any effects in Alzheimer's disease (AD) models. We used H. serrata collected in Japan and determined its huperzine A content using HPLC. We determined its inhibitory effects on acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activity. H. serrata extract (30 mg/kg/day) and donepezil (10 mg/kg/day) were orally administrated for 7 days. After repeated administration, we performed the Y-maze and passive avoidance tests. H. serrata extract contained 0.5% huperzine A; H. serrata extract inhibited AChE, but not BuChE. H. serrata extract ameliorated cognitive function in mice. These results indicate that Japanese H. serrata extract ameliorates cognitive function deficits by inhibiting AChE. Therefore, H. serrata extract may be valuable for the prevention or treatment of dementia in AD.
Assuntos
Alcaloides/administração & dosagem , Inibidores da Colinesterase/administração & dosagem , Transtornos Cognitivos/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Sesquiterpenos/administração & dosagem , Acetilcolinesterase/biossíntese , Acetilcolinesterase/efeitos dos fármacos , Alcaloides/química , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/patologia , Animais , Butirilcolinesterase/biossíntese , Butirilcolinesterase/efeitos dos fármacos , Transtornos Cognitivos/induzido quimicamente , Transtornos Cognitivos/patologia , Huperzia/química , Japão , Transtornos da Memória/tratamento farmacológico , Transtornos da Memória/patologia , Camundongos , Extratos Vegetais/química , Escopolamina/toxicidade , Sesquiterpenos/químicaRESUMO
BACKGROUND: Blue light is a high-energy or short-wavelength visible light, which induces retinal diseases such as age-related macular degeneration and retinitis pigmentosa. Bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea) contain high amounts of polyphenols (anthocyanins, resveratrol, and proanthocyanidins) and thus confer health benefits. This study aimed to determine the protective effects and mechanism of action of bilberry extract (B-ext) and lingonberry extract (L-ext) and their active components against blue light-emitting diode (LED) light-induced retinal photoreceptor cell damage. METHODS: Cultured murine photoreceptor (661 W) cells were exposed to blue LED light following treatment with B-ext, L-ext, or their constituents (cyanidin, delphinidin, malvidin, trans-resveratrol, and procyanidin B2). 661 W cell viability was assessed using a tetrazolium salt (WST-8) assay and Hoechst 33342 nuclear staining, and intracellular reactive oxygen species (ROS) production was determined using CM-H2DCFDA after blue LED light exposure. Activation of p38 mitogen-activated protein kinase (p38 MAPK), nuclear factor-kappa B (NF-κB), and LC3, an ubiquitin-like protein that is necessary for the formation of autophagosomes, were analyzed using Western blotting. Caspase-3/7 activation caused by blue LED light exposure in 661 W cells was determined using a caspase-3/7 assay kit. RESULTS: B-ext, L-ext, NAC, and their active components improved the viability of 661 W cells and inhibited the generation of intracellular ROS induced by blue LED light irradiation. Furthermore, B-ext and L-ext inhibited the activation of p38 MAPK and NF-κB induced by blue LED light exposure. Finally, B-ext, L-ext, and NAC inhibited caspase-3/7 activation and autophagy. CONCLUSIONS: These findings suggest that B-ext and L-ext containing high amounts of polyphenols exert protective effects against blue LED light-induced retinal photoreceptor cell damage mainly through inhibition of ROS production and activation of pro-apoptotic proteins.
Assuntos
Luz , Células Fotorreceptoras de Vertebrados/efeitos dos fármacos , Fitoterapia , Polifenóis/farmacologia , Doenças Retinianas/metabolismo , Vaccinium myrtillus/química , Vaccinium vitis-Idaea/química , Animais , Antocianinas/química , Antocianinas/farmacologia , Antocianinas/uso terapêutico , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/metabolismo , Caspase 3/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Técnicas In Vitro , Camundongos , NF-kappa B/metabolismo , Células Fotorreceptoras de Vertebrados/metabolismo , Células Fotorreceptoras de Vertebrados/efeitos da radiação , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Polifenóis/uso terapêutico , Espécies Reativas de Oxigênio/metabolismo , Resveratrol , Retina/efeitos dos fármacos , Retina/metabolismo , Retina/efeitos da radiação , Doenças Retinianas/tratamento farmacológico , Doenças Retinianas/etiologia , Estilbenos/farmacologia , Estilbenos/uso terapêutico , Sais de Tetrazólio , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Bilberry extract (B-ext) and lingonberry extract (L-ext) are currently used as health supplements. We investigated the protective mechanisms of the B-ext and L-ext against ultraviolet A (UVA)-induced retinal photoreceptor cell damage. Cultured murine photoreceptor (661W) cells were exposed to UVA following treatment with B-ext and L-ext and their main constituents (cyanidin, delphinidin, malvidin, trans-resveratrol, and procyanidin). B-ext, L-ext, and constituents improved cell viability and suppressed ROS generation. Phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK), c-Jun N-terminal kinase (JNK), and protein kinase B (Akt) were analyzed by Western blotting. B-ext and cyanidin inhibited phosphorylation of p38 MAPK, and B-ext also inhibited phosphorylation of JNK by UVA. L-ext, trans-resveratrol, and procyanidin alleviated the reduction of phosphorylated Akt levels by UVA. Finally, a cotreatment with B-ext and L-ext showed an additive effect on cell viability. Our findings suggest that both B-ext and L-ext endow protective effects against UVA-induced retinal damage.
Assuntos
Células Fotorreceptoras de Vertebrados/efeitos dos fármacos , Células Fotorreceptoras de Vertebrados/efeitos da radiação , Extratos Vegetais/farmacologia , Substâncias Protetoras/farmacologia , Raios Ultravioleta/efeitos adversos , Vaccinium myrtillus/química , Vaccinium vitis-Idaea/química , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Células Fotorreceptoras de Vertebrados/citologia , Células Fotorreceptoras de Vertebrados/enzimologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
The purpose of this study was to establish an experimental glaucoma model in the common marmoset (Callithrix jacchus). Chronic intraocular pressure (IOP) elevation was induced by laser trabeculoplasty twice at 2-week intervals in the left eyes of 4 common marmosets. IOP was measured before and at 4, 7, 8, 11, 13 weeks after first laser treatment, and ophthalmoscopic examinations were also performed. At 13 weeks after laser treatment, each eye was enucleated, and retinal cross-sections and optic nerve were prepared for histological examination. Mean IOP values measured at the above 5 time points were over 40 mmHg in laser-treated eyes in 3 marmosets, but IOP in one marmoset was transiently increased to 26.6 mmHg at 7 weeks and then declined to the baseline level. In ophthalmoscopy, deepened and enlarged optic disc cupping, depending on the extent of IOP elevation and duration, were observed in laser-treated eyes of 3 marmosets with persistent IOP elevation, but there was no apparent change in the optic disc in the laser-treated eye of one marmoset with transient IOP elevation. Histological examination showed marked atrophy with deepened and enlarged cupping of optic disc, thinning of retinal nerve fiber layer and retinal ganglion loss in the retina, and axonal atrophy and loss in the optic nerve, depending on the extent of IOP elevation and duration. In conclusion, we succeeded in producing an experimental glaucoma model in the common marmoset, and this model may be useful in elucidating the pathophysiological mechanism for glaucoma.
Assuntos
Callithrix , Modelos Animais de Doenças , Pressão Intraocular/fisiologia , Hipertensão Ocular/fisiopatologia , Hipertensão Ocular/terapia , Animais , Coagulação com Plasma de Argônio , Feminino , Gonioscopia , Lasers de Gás , Hipertensão Ocular/patologia , Oftalmoscopia , Disco Óptico/patologia , Disco Óptico/fisiopatologia , Degeneração Retiniana/patologia , Degeneração Retiniana/fisiopatologia , Malha Trabecular/patologia , Malha Trabecular/fisiopatologia , TrabeculectomiaRESUMO
AIMS: Endoplasmic reticulum (ER) stress has been implicated as a cause of various neurodegenerative diseases. We evaluated the protective effects of purple rice (Oryza sativa L.) bran extract (PRE) and its constituents, namely cyanidin, peonidin, and a newly isolated compound 2-hydroxy-5-[(3S)-3-hydroxybutyl]phenyl-ß-D-glucoside (HHPG), against tunicamycin-induced retinal damage. MAIN METHODS: In an in vitro experiment, protective effects of PRE, cyanidin and HHPG on cultured retinal ganglion cells (RGC-5), which were damaged by treatment with H(2)O(2) or tunicamycin for 24 h, were evaluated. We also investigated the underlying mechanism by examining expressions of ER stress-related proteins, such as immunoglobulin heavy-chain binding protein (BiP) and C/EBP homologous protein (CHOP), and activation of caspase-3 induced by tunicamycin. In an in vivo experiment, mice retinal damage was induced by intravitreous injection of tunicamycin as revealed by histological analysis using hematoxylin-eosin staining. KEY FINDINGS: The viability of H(2)O(2) or tunicamycin-treated RGC-5, assessed using the tetrazolium salt (WST-8) assay, was improved by treatment with PRE, cyanidin, and HHPG, respectively. PRE did not affect tunicamycin-induced expressions of BiP or CHOP. However, PRE, cyanidin, and HHPG suppressed tunicamycin-induced caspase-3 activation. Histological analysis using hematoxylin-eosin staining showed that intravitreous injection of PRE significantly suppressed the tunicamycin-induced degeneration of retinal ganglion cells in mice. SIGNIFICANCE: These findings indicate that PRE, cyanidin, and HHPG suppress tunicamycin-induced retinal ganglion cell death at least partly by inhibiting activation of caspase-3, suggesting that PRE and its main constituents prevent retinal disease caused by ER stress.
Assuntos
Morte Celular/efeitos dos fármacos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Glucosídeos/química , Oryza/química , Extratos Vegetais/farmacologia , Células Ganglionares da Retina/efeitos dos fármacos , Animais , Antocianinas/isolamento & purificação , Antocianinas/farmacologia , Caspase 3/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Glucosídeos/isolamento & purificação , Glucosídeos/farmacologia , Peróxido de Hidrogênio/toxicidade , Masculino , Camundongos , Células Ganglionares da Retina/patologia , Tunicamicina/toxicidadeRESUMO
Background. We investigated the effects of Brazilian green propolis and its constituents against white light- or UVA-induced cell damage in mouse retinal cone-cell line 661W or human skin-derived fibroblast cells (NB1-RGB). Methods. Cell damage was induced by 3,000lx white light for 24 h or 4/10 J/cm(2) UVA exposure. Cell viability was assessed by Hoechst33342 and propidium iodide staining or by tetrazolium salt (WST-8) cell viability assay. The radical scavenging activity of propolis induced by UVA irradiation in NB1-RGB cells was measured using a reactive-oxygen-species- (ROS-) sensitive probe CM-H2DCFDA. Moreover, the effects of propolis on the UVA-induced activation of p38 and extracellular signal-regulated kinase (ERK) were examined by immunoblotting. Results. Treatment with propolis and two dicaffeoylquinic acids significantly inhibited the decrease in cell viability induced by white light in 661W. Propolis and its constituents inhibited the decrease in cell viability induced by UVA in NB1-RGB. Moreover, propolis suppressed the intracellular ROS production by UVA irradiation. Propolis also inhibited the levels of phosphorylated-p38 and ERK by UVA irradiation. Conclusion. Brazilian green propolis may become a major therapeutic candidate for the treatment of AMD and skin damage induced by UV irradiation.
RESUMO
BACKGROUND: Brazilian green propolis is reported to have wide range of biological properties including antibacterial, anti-inflammatory, anti-influenza, and antioxidant activities. In the digestive system, a protective effect of propolis on gastric ulcer has been reported, but a laxative effect has not yet been reported. We investigated the effect and the mechanism of action of water and ethanol extracts of Brazilian green propolis. METHODS: We examined the laxative effect of propolis on stool frequency by administering orally an ethanol extract of propolis (EEP) or a water extract of propolis (WEP) at 10, 50, 100, or 500 mg/kg to normal mice. We then investigated the effects of propolis using constipation model mice induced by two types of drugs, loperamide (a µ opioid receptor agonist) and clonidine (an α-2 adrenergic receptor agonist). We also investigated the effects of WEP on gastrointestinal transit and contractional tension of the ileum to uncover the mechanism of action of WEP. RESULTS: Treatment with WEP, but not with EEP, significantly increased the weight of stools (p<0.01 at 500 mg/kg). WEP treatment significantly restored stool frequency and stool weight in clonidine-induced constipation model mice, but not in loperamide-induced constipation model mice. WEP treatment did not affect gastro-intestinal transit, but significantly increased the contractional tension of the isolated ileum of guinea pigs. This increase was inhibited by an acetylcholine receptor antagonist (atropine), but not by a 5-HT receptor antagonist (GR113808). CONCLUSION: These findings indicate that WEP has laxative effects both in normal mice and in clonidine-induced constipation model mice. The laxative effects of WEP might be mediated by increased contractional tension of the ileum exerted at least in part via activation of an acetylcholine receptor.
Assuntos
Constipação Intestinal/tratamento farmacológico , Laxantes/administração & dosagem , Própole/administração & dosagem , Animais , Abelhas , Brasil , Constipação Intestinal/fisiopatologia , Trânsito Gastrointestinal/efeitos dos fármacos , Cobaias , Humanos , Masculino , CamundongosRESUMO
Crocetin, an aglycone of crocin, is found in stigmas of the saffron crocus (Crocus starus L.) and has been used in traditional medicine. We investigated the effects of oral administration of crocetin on damage induced by N-methyl-D-aspartate (NMDA) in the murine retina. Crocetin was orally administered before and after intravitreal injection of NMDA. A histological analysis was conducted by counting the cell number of ganglion cell layer (GCL). Cell apoptosis was assessed by counting cells positive for terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Retinal functions were measured in terms of a- and b-wave amplitudes using an electroretinogram (ERG). Activation of caspase-3/7 and cleaved caspase-3 expression were assayed. Calpain activity was evaluated by immunoblotting assays for proteolysis of α-spectrin. NMDA injection decreased the cell number in the GCL, and crocetin at a dose of 100 mg/kg inhibited this reduction. TUNEL-positive cells were observed in both GCL and inner nuclear layer (INL) after NMDA injection, and crocetin inhibited the increase in number of TUNEL-positive cells. ERG analysis showed that both a- and b-wave amplitudes were decreased by NMDA injection. Crocetin inhibited the reduction in the b-wave amplitude, but not in the a-wave. NMDA injection activated caspase-3/7 and increased expression of cleaved caspsase-3 in the GCL and INL, but both of these processes were inhibited by crocetin. NMDA injection also induced cleavage of α-spectrin, but crocetin did not affect this process. These findings indicate that oral administration of crocetin prevented NMDA-induced retinal damage via inhibition of the caspase pathway.
Assuntos
Carotenoides/administração & dosagem , N-Metilaspartato/efeitos adversos , Retina/efeitos dos fármacos , Retina/lesões , Administração Oral , Animais , Apoptose/efeitos dos fármacos , Calpaína/biossíntese , Caspase 3/biossíntese , Contagem de Células , Citoproteção/efeitos dos fármacos , Indução Enzimática/efeitos dos fármacos , Masculino , Camundongos , Retina/metabolismo , Retina/patologia , Células Ganglionares da Retina/efeitos dos fármacos , Células Ganglionares da Retina/metabolismo , Células Ganglionares da Retina/patologia , Vitamina A/análogos & derivadosRESUMO
SCOPE: Annatto (Bixa orellana) seeds have been used as a colorant in butter and in a variety of other foods. In this study, we investigated the amelioration of retinal damage by an acetone extract of annatto (A-ext.), bixin (a main component of annatto), and four bixin derivatives (Bx-1, Bx-2, Bx-3, and Bx-4) that we have synthesized. METHODS AND RESULTS: We used cultured retinal ganglion cells (RGC-5) to examine in vitro effects of A-ext. on stress pathways, focusing on intracellular oxidation induced by reactive oxygen species, expression of endoplasmic reticulum (ER) stress-related proteins, caspase-3 activation, and cell membrane damage. In vivo retinal damage in mice following intravitreous injection of tunicamycin was evaluated by counting the cell numbers in the ganglion cell layer (GCL) and measuring the thickness of outer nuclear layer (ONL). A-ext., bixin, and Bx-1 treatment inhibited both tunicamycin- and H2O2-induced cell death. Bixin derivatives also inhibited tunicamycin-induced cell death. Treatment with A-ext., bixin, and Bx-1 reduced tunicamycin-induced caspase-3 activity and inhibited the inversion of phosphatidylserine, an early apoptotic event without antioxidant effect or reduction of ER stress itself. A-ext., bixin, and Bx-1 significantly inhibited the tunicamycin-induced loss of cells from the GCL, and these materials also suppressed the tunicamycin-induced thinning of ONL. CONCLUSION: A-ext., its main component bixin, and bixin derivatives may therefore be useful for preventive and therapeutic treatment of retinal-related diseases.
Assuntos
Carotenoides/farmacologia , Estresse do Retículo Endoplasmático , Extratos Vegetais/farmacologia , Degeneração Retiniana/prevenção & controle , Animais , Bixaceae , Caspase 3/metabolismo , Morte Celular/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Células Cultivadas , Avaliação Pré-Clínica de Medicamentos/métodos , Peróxido de Hidrogênio/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos , Fosfatidilserinas/metabolismo , Degeneração Retiniana/etiologia , Células Ganglionares da Retina/efeitos dos fármacos , Tunicamicina/efeitos adversosRESUMO
The study evaluated the protective effects of purple rice (Oryza sativa L.) bran extract (PRE) and its constituents, cyanidin and peonidin, against angiogenesis induced by vascular endothelial growth factor (VEGF). The effects of VEGF and PRE were examined by in vitro tube formation assays and following 14-day co-culture of human umbilical vein endothelial cells (HUVECs) and fibroblasts. The antiangiogenic mechanism of PRE was evaluated by VEGF-induced proliferation and migration of HUVECs and/or human retinal microvascular endothelial cells (HRMECs) and phosphorylation of extracellular signal-regulated kinase (ERK) and p38. The PRE significantly suppressed VEGF-induced tube formation, proliferation and migration in HUVECs and HRMECs as well as phosphorylation of ERK and p38. Cyanidin and peonidin also suppressed the proliferation and migration induced by VEGF. These findings indicate that PRE and anthocyanidins suppress VEGF-induced angiogenesis by inhibiting proliferation and migration and suggest that the inhibition of phosphorylated-ERK and -p38 may be involved in the underlying mechanism.
Assuntos
Inibidores da Angiogênese/farmacologia , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Oryza/química , Extratos Vegetais/farmacologia , Antocianinas/farmacologia , Células Cultivadas , Técnicas de Cocultura , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fibroblastos/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Fosforilação , Retina/citologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
The purpose of this study was to investigate the effect of astaxanthin extracted from Paracoccus carotinifaciens on gastric mucosal damage in murine gastric ulcer models. Mice were pretreated with astaxanthin for 1 h before ulcer induction. Gastric ulcers were induced in mice by oral administration of hydrochloride (HCl)/ethanol or acidified aspirin. The effect of astaxanthin on lipid peroxidation in murine stomach homogenates was also evaluated by measuring the level of thiobarbituric acid reactive substance (TBARS). The free radical scavenging activities of astaxanthin were also measured by electron spin resonance (ESR) measurements. Astaxanthin significantly decreased the extent of HCl/ethanol- and acidified aspirin-induced gastric ulcers. Astaxanthin also decreased the level of TBARS. The ESR measurement showed that astaxanthin had radical scavenging activities against the 1,1-diphenyl-2-picrylhydrazyl radical and the superoxide anion radical. These results suggest that astaxanthin has antioxidant properties and exerts a protective effect against ulcer formation in murine models.
Assuntos
Antiulcerosos/uso terapêutico , Paracoccus/química , Úlcera Gástrica/tratamento farmacológico , Ácidos/administração & dosagem , Ácidos/efeitos adversos , Animais , Antiulcerosos/administração & dosagem , Aspirina/administração & dosagem , Aspirina/efeitos adversos , Compostos de Bifenilo/metabolismo , Modelos Animais de Doenças , Etanol/administração & dosagem , Etanol/efeitos adversos , Sequestradores de Radicais Livres/administração & dosagem , Sequestradores de Radicais Livres/uso terapêutico , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Peroxidação de Lipídeos , Masculino , Camundongos , Picratos/metabolismo , Estômago/efeitos dos fármacos , Estômago/patologia , Úlcera Gástrica/induzido quimicamente , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Xantofilas/administração & dosagem , Xantofilas/química , Xantofilas/uso terapêuticoRESUMO
This study evaluated the protective effects of purple rice ( Oryza sativa L.) bran extract (PRE) and its major anthocyanidins (cyanidin and peonidin) against light-induced retinal damage. In an in vitro experiment, cultured murine photoreceptor cells (661W) were damaged by a 24 h exposure to light. Viability of 661W after light treatment, assessed by the tetrazolium salt (WST-8) assay and Hoechst 33342 nuclear staining, was improved by the addition of PRE, cyanidin, and peonidin. Intracellular radical activation in 661W, evaluated using the reactive oxygen species (ROS) sensitive probe 5-(and 6)-chloromethyl-2,7-dichlorodihydrofluorescein diacetate acetyl ester (CM-H(2)DCFDA), was reduced by PRE and its anthocyanidins. Electron spin resonance (ESR) measurements showed that PRE, peonidin, and cyanidin all exhibited radical scavenging activities against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, superoxide anion radical ((â¢)O(2)(-)), and hydroxyl radical ((â¢)OH). In an in vivo mouse experiment, intravitreous injection of PRE significantly suppressed photoreceptor degeneration induced by exposure to light as revealed by histological analysis using hematoxylin-eosin staining. These findings suggest that PRE and its anthocyanidins possess protective effects with antioxidation mechanism in both in vitro and in vivo models of retinal diseases.
Assuntos
Antocianinas/farmacologia , Oryza/química , Extratos Vegetais/farmacologia , Substâncias Protetoras/farmacologia , Retina/efeitos da radiação , Doenças Retinianas/tratamento farmacológico , Animais , Antocianinas/administração & dosagem , Modelos Animais de Doenças , Humanos , Luz , Masculino , Camundongos , Extratos Vegetais/administração & dosagem , Substâncias Protetoras/administração & dosagem , Retina/efeitos dos fármacos , Retina/lesõesRESUMO
BACKGROUND: Agarwood (Aquilaria sinensis), well known as incense in Southeast Asia, has been used as a digestive in traditional medicine. We investigated the laxative effects of an ethanol extract of agarwood leaves (EEA) in a rat model of low-fiber diet-induced constipation. METHODS: A set of rats was bred on a normal diet while another set was placed on a low-fiber diet to induce constipation. The laxative effect of agarwood was then investigated on both sets of rats. RESULTS: Pretreatment of normal rats with single dose of EEA (600 mg/kg, p.o.) significantly increased frequency and weight of stools. Also, treatments with EEA (300 and 600 mg/kg, p.o.) for 14 days caused a significant increase in stool frequency and weight. Feeding of the animals with a low-fiber diet resulted in a decrease in stool weight, frequency, and water content and also delayed carmine egestion. A single treatment with EEA (600 mg/kg) or senna (150 and 300 mg/kg) significantly increased stool frequency, weight, and water content and also accelerated carmine egestion in the model rats. Once daily administrations of EEA (150 mg/kg), for 14 days, caused a significant increase in water content of stools. The higher doses of EEA (300 and 600 mg/kg) significantly increased frequency, weight, and water content of the stools while accelerating carmine egestion in the constipated rats. Senna (150 and 300 mg/kg) produced similar effect as the higher doses of EEA but, in addition, induced severe diarrhea. CONCLUSION: These findings indicate that EEA has a laxative effect, without causing diarrhea, in a rat model of low-fiber diet-induced constipation. These findings suggest that EEA may be highly effective on constipation as a complementary medicine in humans suffering from life style-induced constipation.
Assuntos
Constipação Intestinal/tratamento farmacológico , Defecação/efeitos dos fármacos , Laxantes/uso terapêutico , Extratos Vegetais/uso terapêutico , Senna , Thymelaeaceae , Animais , Carmim/análise , Constipação Intestinal/etiologia , Diarreia , Dieta , Fibras na Dieta/administração & dosagem , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Fezes/química , Laxantes/farmacologia , Masculino , Extratos Vegetais/farmacologia , Folhas de Planta , Ratos , Ratos Sprague-Dawley , Água/análiseRESUMO
Agarwood (Aquilaria sinensis, Aquilaria crasna) is well known as an incense in the oriental region such as Thailand, Taiwan, and Cambodia, and is used as a digestive in traditional medicine. We investigated the laxative effects and mechanism of agarwood leaves extracted with ethanol (EEA-1, Aquilaria sinensis; EEA-2, Aquilaria crasna). EEA-1, EEA-2, the main constituents of EEAs (mangiferin, and genkwanin-5-O-primeveroside), and senna increased the frequency and weight of stools in loperamide-induced constipation model mice. EEA-1 and EEA-2 did not induce diarrhea as a side effect, but senna induced severe diarrhea. EEA-1 and senna increased gastro-intestinal (GI) transit in the model mice. EEA-1, but not senna, also increased the intestinal tension of isolated jejunum and ileum in guinea pigs, and the tension increase was blocked by atropine, a muscarinic receptor antagonist, but not by other inhibitors (granicetron, pyrilamine, or bradykinin-antagonist peptide). Furthermore, the increase in frequency and weight of stools induced by EEA-1 were blocked by pre-administration of atropine in the model mice. These findings indicate that EEAs exerted a laxative effect via acetylcholine receptors in the mouse constipation model.
Assuntos
Constipação Intestinal/induzido quimicamente , Constipação Intestinal/tratamento farmacológico , Laxantes/farmacologia , Loperamida/farmacologia , Extratos Vegetais/farmacologia , Receptores Colinérgicos/metabolismo , Thymelaeaceae/química , Animais , Atropina/farmacologia , Antagonistas Colinérgicos/farmacologia , Constipação Intestinal/metabolismo , Diarreia/induzido quimicamente , Etanol/química , Feminino , Trânsito Gastrointestinal/efeitos dos fármacos , Cobaias , Intestinos/efeitos dos fármacos , Intestinos/fisiopatologia , Laxantes/efeitos adversos , Laxantes/uso terapêutico , Masculino , Camundongos , Extratos Vegetais/efeitos adversos , Extratos Vegetais/uso terapêutico , Folhas de Planta/químicaRESUMO
The aim of this study was to examine the antiangiogenic properties and antioxidant activities (a) of the main anthocyanidins (delphinidin, cyanidin and malvidin) found as constituents in Vaccinium myrtillus (bilberry) anthocyanosides (VMA) and (b) of N-acetyl-L-cysteine (NAC). Each of these anthocyanidins concentration-dependently inhibited vascular endothelial growth factor (VEGF)-induced tube formation in a co-culture of human umbilical vein endothelial cells (HUVECs) and fibroblasts, the effect of each anthocyanidin being significant at 3 and/or 10 microM, while NAC significantly inhibited such tube formation at 1 microM (the only concentration tested). Moreover, each anthocyanidin (0.3-10 microM) and NAC (1-1000 microM) concentration-dependently scavenged the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. The inhibitory effects against angiogenesis were similar among the anthocyanidins, as were those against the DPPH radical. Moreover, their radical-scavenging effects were induced by concentrations that were at or below those that induced their antiangiogenic effects. These findings indicate that the inhibitory effect of VMA on angiogenesis may depend on those of its main constituent anthocyanidins (delphinidin, cyanidin and malvidin), presumably via antioxidant effects.