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1.
Int J Mol Med ; 45(2): 343-352, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31789424

RESUMO

Oxidative and inflammatory damage has been suggested to play important roles in the pathogenesis of skin photoaging. Andrographolide sodium bisulfate (ASB) is a soluble derivative of andrographolide and has known antioxidant and anti­inflammatory properties. In the present study, cellular experiments were designed to investigate the molecular mechanisms underlying the effect of ASB in relieving ultraviolet (UV)­induced photo­damage. Following ASB pretreatment and UV irradiation, the apoptosis and necrosis of HaCaT cells were investigated by Hoechst 33342/propidium iodide staining. Reactive oxygen species (ROS) production was investigated using a DCFH­DA fluorescence probe. Furthermore, the protein expression levels of p65, NF­κB inhibitor­α, nuclear factor E2­related factor 2 (Nrf2) and kelch­like ECH­associated protein 1 (keap1) were measured via western blotting and immunofluorescence analyses. Furthermore, NF­κB­mediated cytokines were assessed by ELISA, and Nrf2­mediated genes were detected by reverse transcription­quantitative PCR. Pretreatment with ASB markedly increased cell viability, decreased cell apoptosis and decreased UV­induced excess ROS levels. In addition, ASB activated the production of Nrf2 and increased the mRNA expression levels of glutamate­cysteine ligase catalytic subunit and NAD(P)H quinone oxidoreductase 1, while ASB downregulated the protein expression of p65 and decreased the production of interleukin (IL)­1ß, IL­6 and tumor necrosis factor­α. These results suggested that ASB attenuates UV­induced photo­damage by activating the keap1/Nrf2 pathway and downregulating the NF­κB pathway in HaCaT keratinocytes.

2.
J Surg Res ; 245: 441-452, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31445496

RESUMO

BACKGROUND: Whitmania pigra Whitman (W pigra), a traditional Chinese medicine, has functions of breaking stagnant and eliminating blood stasis. The aim of this study was to investigate the underlying mechanism of W pigra against deep vein thrombosis (DVT). METHODS: A rat model of DVT induced by inferior vena cava stenosis was successfully established. Rats were administered vehicle (saline solution, p.o.), three doses of W pigra aqueous extract (34.7, 104.2, or 312.5 mg crude W pigra/kg, p.o.), heparin (200 U/kg, i.v.), or clopidogrel (25 mg/kg, p.o.) once daily for 2 d. Thrombus weight and histopathological changes were examined. Blood samples were collected to determine blood cell counts, blood viscosity, blood coagulation, blood fibrinolysis, serum levels of interleukin-1ß, and tumor necrosis factor-α. Protein expressions of Sirtuin1 (SIRT1), acetylated p65 (Ace-p65), and phosphorylated p65 (p-p65) were determined by Western blot. Furthermore, SIRT1-specific inhibitor EX527 was applied to confirm the role of SIRT1 in the antithrombotic effect of W pigra. RESULTS: W pigra significantly decreased thrombus weight. W pigra had no effects on blood cell counts, whole blood viscosity, blood coagulation, blood fibrinolysis. However, it reduced tissue factor protein expression in the vein wall and thrombus. Moreover, it sharply increased SIRT1 protein expression and decreased leukocytes recruitment in the thrombus and vein wall, serum levels of interleukin-1ß and tumor necrosis factor-α, and protein expressions of Ace-p65 and p-p65. Furthermore, the antithrombotic effect of W pigra was significantly abolished by EX527. CONCLUSIONS: Aqueous extract of W pigra effectively reduced DVT burden by inhibiting inflammation via SIRT1/nuclear factor-kappa B signaling pathway.


Assuntos
Produtos Biológicos/uso terapêutico , Sanguessugas , NF-kappa B/metabolismo , Sirtuína 1/metabolismo , Trombose Venosa/tratamento farmacológico , Animais , Produtos Biológicos/farmacologia , Carbazóis , Citocinas/sangue , Avaliação Pré-Clínica de Medicamentos , Feminino , Inflamação/tratamento farmacológico , Masculino , Medicina Tradicional Chinesa , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Sirtuína 1/antagonistas & inibidores , Tromboplastina/metabolismo , Trombose Venosa/metabolismo
3.
Am J Chin Med ; 44(4): 755-69, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27222060

RESUMO

This study aimed to investigate the effect of timosaponin B-II (TB-II) on palmitate (PA)-induced insulin resistance and inflammation in HepG2 cells, and probe the potential mechanisms. TB-II, a main ingredient of the traditional Chinese medicine Anemarrhena asphodeloides Bunge, notably ameliorated PA-induced insulin resistance and inflammation, and significantly improved cell viability, decreased PA-induced production of tumor necrosis factor-[Formula: see text] (TNF-[Formula: see text]) and interleukin-6 (IL-6) levels. Further, TB-II treatment notably decreased malondialdehyde (MDA) and lactate dehydrogenase (LDH) levels, and improved superoxide dismutase (SOD) and nitric oxide (NO). TB-II also reduced HepG2 cells apoptosis. Insulin receptor substrate-1 (IRS1)/phosphatidylinositol 3-kinase (PI3K)/Akt and inhibitor of nuclear factor [Formula: see text]-B kinase (IKK)/NF-[Formula: see text]B pathways-related proteins, and IKK[Formula: see text], p65 phosphorylation, serine phosphorylation of insulin receptor substrate-1 (IRS-1) at S307, tyrosine phosphorylation of IRS-1, and Akt activation were determined by Western blot. Compared to model group, TB-II significantly downregulated the expression of p-NF-[Formula: see text]Bp65, p-IKK[Formula: see text], p-IRS-1, p-PI3K and p-Akt. TB-II is a promising potential agent for the management of palmitate-induced insulin resistance and inflammation, which might be via IR/IRS-1/PI3K/Akt and IKK/NF-[Formula: see text]B pathways.


Assuntos
Inflamação/tratamento farmacológico , Proteínas Substratos do Receptor de Insulina/metabolismo , Resistência à Insulina , NF-kappa B/metabolismo , Palmitatos/efeitos adversos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Saponinas/farmacologia , Esteroides/farmacologia , Células Hep G2 , Humanos , Quinase I-kappa B , Inflamação/genética , Inflamação/metabolismo , Proteínas Substratos do Receptor de Insulina/genética , Interleucina-6/genética , Interleucina-6/metabolismo , NF-kappa B/genética , Óxido Nítrico/metabolismo , Fosfatidilinositol 3-Quinases/genética , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais/efeitos dos fármacos
4.
Int J Ophthalmol ; 8(6): 1094-100, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26682154

RESUMO

AIM: To evaluate effects of Danhong Huayu Koufuye (DHK, a Chinese medicinal formulae) alone or combined with metformin on diabetic retinopathy (DR) in Zucker diabetic fatty (ZDF) rats, an animal model of obese type-2 diabetes, and then to investigate the mechanisms. METHODS: ZDF (fa/fa) rats were administered with vehicle (distilled water), metformin, DHK, and DHK plus metformin. Electrophysiological and histological analysis were applied to evaluated effects of DHK alone or combined with metformin on DR. The levels of fasting blood glucose (FBG) and glycosylated hemoglobin (HbA1c) in blood were measured to evaluate the antihyperglycemic activity of DHK. Furthermore, levels of nitric oxide (NO), malondialdehyde (MDA) and activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) in serum were measured to study effects of DHK on oxidative stress in ZDF rats. In addition, body weight, lipidic indexes and insulin level were also assessed. RESULTS: DHK combined with metformin significantly reversed the prolongation of latency times of flash electroretinogram (FERG) and oscillatory potentials (OPs) in diabetic rats. Furthermore, DHK alone or combined with metformin showed a remarkable suppression of retinal neovascularization and amelioration of retinal internal limiting membrane morphology. Moreover, DHK alone or plus metformin reduced FBG (P<0.05), HbA1c (P<0.01) and MDA (P<0.01) levels in diabetic rats. In addition, reductions in levels of triglycerides (TG) (P<0.01) and low density lipoprotein cholesterol (LDL-c) (P<0.01 and P<0.05, respectively) were also observed in diabetic rats treated with DHK alone or plus metformin. CONCLUSION: DHK in combination with metformin had a preventive and therapeutic effect on DR in type-2 diabetic rats, and the possible mechanisms may be alleviating hyperglycemia, reducing oxidative stress and improving lipid metabolism.

5.
Int J Ophthalmol ; 7(3): 391-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24967179

RESUMO

AIM: To investigate the effects of naringenin eye drops on N-methyl-N-nitrosourea (MNU)-induced photoreceptor cell death in rats. METHODS: Photoreceptor cell death was induced by single intraperitoneal injection of MNU (60 mg/kg) in rats. Both eyes of all animals were instilled with one drop of vehicle, 0.5% or 1.0% naringenin eye drops three times per day from 7d before to 17d after MNU injection. Effects of naringenin on MNU-induced photoreceptor cell death were evaluated by electrophysiological and histological analysis. RESULTS: Flash electroretinography (FERG) and oscillatory potentials (OPs) recordings showed that the vehicle control group had remarkable reduction of amplitudes and prolongation of latency times. FERG and OPs responses were significantly reversed in MNU-induced rats treated with 0.5% or 1.0% naringenin eye drops compared with the vehicle control. The retinal morphological results showed that naringenin dose-dependently preserved the outer nuclear layer, outer retina and total retina. CONCLUSION: These results indicate that topical treatment with naringenin eye drops prevented retinal neurons from MNU-induced structural and functional damages.

6.
J Ethnopharmacol ; 153(3): 793-800, 2014 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-24685587

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Genkwa flos (Daphne genkwa Sieb. et Zucc.), a Chinese herbal medicine, has been traditionally used for over two thousand years in China for inflammation related symptoms, including joint pain. To evaluate the antioxidative effects of flavonoid aglycones (FA) isolated from Genkwa flos on adjuvant arthritis in rats and to identify the relationship between antioxidant potential and whole blood viscosity (WBV). MATERIALS AND METHODS: FA compounds were identified using LC-MS and the content was assayed by HPLC. Arthritis was induced by an intradermal injection of Freund׳s complete adjuvant in the footpad. The effects of FA on paw volumes, secondary arthritis scores, histopathology of joints, and body and organ weights were measured. The antioxidant effects of FA and WBV were determined. RESULTS: LC-MS analysis showed that the FA contained four major compounds: luteolin, apigenin, hydroxygenkwanin and genkwanin. FA significantly decreased paw edema, arthritis scores, and weight loss. These observations were consistent with the reduction of oxidative stress and the improvement of the WBV. CONCLUSION: FA significantly decreased arthritis in a rat model through antioxidant and hemorheological modulatory mechanisms. The Genkwa flos flavonoids may have clinical potential for the treatment of rheumatoid arthritis.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Artrite Experimental/metabolismo , Daphne , Flavonoides/farmacologia , Animais , Articulação do Tornozelo/efeitos dos fármacos , Articulação do Tornozelo/patologia , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/uso terapêutico , Antioxidantes/isolamento & purificação , Antioxidantes/uso terapêutico , Artrite Experimental/tratamento farmacológico , Artrite Experimental/patologia , Daphne/química , Flavonoides/isolamento & purificação , Flavonoides/uso terapêutico , Flores/química , Glutationa Peroxidase/metabolismo , Articulação do Joelho/efeitos dos fármacos , Articulação do Joelho/patologia , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Malondialdeído/metabolismo , Óxido Nítrico/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Fitoterapia , Extratos Vegetais/química , Ratos Sprague-Dawley , Baço/efeitos dos fármacos , Baço/patologia , Superóxido Dismutase/metabolismo , Timo/efeitos dos fármacos , Timo/patologia
7.
Zhong Yao Cai ; 37(7): 1218-21, 2014 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-25566660

RESUMO

OBJECTIVE: To investigate effects of Danhong Huayu Koufuye (DHK), insulin and their combination on diabetic cardiomyopathy (DC) in streptozotocin (STZ, 50 mg/kg, ip)-induced diabetic rats. METHODS: Rats were divided into five groups: normal control, diabetic treated with vehicle, insulin, DHK, and DHK plus insulin. The animals were treated once daily for 15 weeks starting one week after STZ injection. RESULTS: The combination of DHK with insulin significantly reduced cardiac index (P < 0.05), serum LDH (P < 0.05), AST(P < 0.05), ALT(P < 0.05) and HDL-C (P < 0.05) level, and promoted pancreatic and cardiac morphological changes as compared with the model group. CONCLUSION: It is suggested that DHK may be a valuable adjuvant therapy for DC.


Assuntos
Cardiomiopatias Diabéticas/tratamento farmacológico , Medicamentos de Ervas Chinesas/química , Insulina/farmacologia , Administração Oral , Envelhecimento , Animais , Glicemia , Diabetes Mellitus Experimental , Progressão da Doença , Pâncreas , Fitoterapia , Ratos , Estreptozocina
8.
J Ethnopharmacol ; 148(3): 818-25, 2013 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-23707329

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Cordyceps sinensis, an entomogenous fungus used in traditional Chinese medicine with multiple pharmacological activities. However, its usage has been limited due to the high price and short supply. Isolate of fungi strains from natural Cordyceps sinensis to achieve a large-scale production by fermentation is an alternative choice. The aim of this study was to investigate and compare the effects of mycelia extracts of different fungal stains isolated from natural Cordyceps sinensis on macrophage functions in vitro. MATERIALS AND METHODS: Macrophages' proliferation, phagocytosis, nitric oxide (NO) production, cytokines secretion, iNOS, NF-κB p65 activation and translocation were investigated by the MTT assay, flow cytometry assay, Griess reagent method, ELISA, western blot and immunostaining assay, respectively. RESULTS: The results showed that the effects of cultured Cordyceps mycelia of different fungal strains isolated from natural Cordyceps sinensis on macrophages greatly variant. Among 17 Cordyceps aqueous extracts, only five extracts (UM01, QH11, BNQM, GNCC and DCXC) could significantly increase cell proliferation and NO production of RAW 264.7 mouse macrophages. Moreover, the five extracts, especially UM01 and QH11, significantly enhanced phagocytosis and promoted cytokines release of macrophages. Polysaccharides in cultured UM01 mycelia were found to be the main immune stimulating compounds. CONCLUSIONS: The variation of biological effects of fermented mycelia of different fungal strains from natural Cordyceps sinensis may be derived from their chemical diversity, especially polysaccharides, which need further study in future.


Assuntos
Produtos Biológicos/farmacologia , Cordyceps , Micélio , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Cordyceps/isolamento & purificação , Citocinas/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Fagocitose/efeitos dos fármacos
9.
Eye Sci ; 26(1): 23-9, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21425492

RESUMO

PURPOSE: Age-related macular degeneration (AMD) as a disease entity is "dry" at early stage and made up of two main components at late stage: atrophic AMD and exudative AMD. Quercetin acts as an anti-oxidant to protect retinal pigment epithelial cells (RPE) from damaged by oxidative stress, but its effect on formation of choroidal neovascularization (CNV) in AMD is unclear. The aim of this study is to investigate the effect of quercetin on the formation of CNV in AMD. METHODS: The development of CNV induced by laser was detected by fluorescein angiography (FA). Colored microsphere technique was used to determine the choroidal blood flow in ocular hypertensive rabbit eyes. In in vitro studies, HUVECs were treated with NaIO3, H2O2 and NaN3 to induce oxidative cell damages. The effect of quercetin on various oxidations-induced injuries in HUVECs was measured by MTT assay. HUVECs migration was assessed using a wound healing assay. RESULTS: Quercetin significantly inhibited the formation of laser-induced CNV. The choroidal blood flow in rabbit eyes was significantly increased after quercetin instillation. In vitro results showed quercetin enhanced various oxidations-induced injuries in HUVECs and inhibited migration of HUVECs during wound healing. CONCLUSION: Quercetin inhibited the formation of CNV both in vivo and in vitro and increased choroidal blood flow. It could become a promising candidate for the treatment of AMD.


Assuntos
Antioxidantes/farmacologia , Neovascularização de Coroide/prevenção & controle , Degeneração Macular/complicações , Quercetina/farmacologia , Animais , Corioide/irrigação sanguínea , Corioide/efeitos dos fármacos , Neovascularização de Coroide/etiologia , Angiofluoresceinografia , Células Endoteliais da Veia Umbilical Humana , Peróxido de Hidrogênio , Iodatos , Lasers , Coelhos , Azida Sódica , Degeneração Macular Exsudativa
10.
Int J Ophthalmol ; 4(6): 599-604, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22553728

RESUMO

AIM: To study the effects of danhong huayu koufuye (DHK) on fasting blood glucose (FBG) and diabetic retinopathy (DR) in streptozotocin (STZ)-induced type 1 diabetic rats to facilitate the rational usage of this drug. METHODS: Diabetic rats were induced by injection of a single dose of STZ intraperitoneally at 50mg/kg. Flash electroretinogram (FERG) and oscillatory potentials (OPs) were used to measure retinal function. The microvascular perfusion of ears was performed to study the microcirculation in rats. FBG, body-weight, and 24-h urine volume, water intake and diet intake were also assessed. RESULTS: DHK had no effect on FBG in normal rats. However, STZ + DHK group were significantly different from those of Model and moved toward those of normal control. It reversed the increase in diet intake (P≤0.05 vs model control) and the loss in body-weight (P≤0.05 vs model control) in diabetic rats. DHK decreased the FBG of diabetic rats by 25.6% (P≤0.05) and 37.9% (P≤0.01) after 14 and 21 days administration as compared with the model control, respectively. Moreover, DHK significantly increased the FERG b-wave amplitude by 80% (P≤0.05 vs model control) and decreased the FERG b-wave latency by 15.3% (P≤0.01 vs model control) after 24 days administration. The OP(1) and OP(2) amplitudes in DHK group were 2.6 (P≤0.01) and 2.0 (P≤0.01) times of model group after 24 days of DHK treatment, respectively. At the same time, OP1 and OP2 latencies in DHK group reduced by 16.0% (P≤0.001) and 14.7% (P≤0.001) as compared with the model control, respectively. Furthermore, the microvascular perfusion of DHK group was 2.4 times of model group (P≤0.001) after 21 days administration. CONCLUSION: DHK had no effect on normal FBG. But it had antihyperglycemic activity, and had a preventive and therapeutic effect on DR in diabetic rats.

11.
Int J Ophthalmol ; 3(3): 205-10, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-22553555

RESUMO

AIM: To study the effects of Tetramethylpyrazine (TMP) on retinal pigment epithelium (RPE) degeneration, choroidal blood flow and oxidative stress of RPE cells. METHODS: The 35mg/kg NaIO(3)-induced RPE degeneration rat eyes was given 25µg 1% TMP eye drops 3 times a day for 7 days before NaIO(3) injection, and then 2 to 4 weeks after NaIO(3) injection. RPE function was measured with c-wave of electroretinogram (ERG). Colored microsphere technique was used for in vivo experiments to determine the choroidal blood flow in ocular hypertensive (40mmHg) rabbit eyes. Methylthiazoltetrazolium (MTT) assay was used to study in vitro effect of TMP on various oxidants induced injury in the hRPE (ARPE-19 (ATCC, Manassas, VA, USA)). RESULTS: Two weeks after NaIO(3) injection, the amplitude of ERG c-wave fell markedly in NaIO(3) group to 36% of control group(P<0.01). No apparent difference was observed in TMP+NaIO(3) group. Four weeks later, the NaIO(3) group fell to 46% of control group (P<0.01), while the TMP+NaIO(3) group fell to only 77% of control group (P<0.01). There was a 67% reversal of the ERG c-wave by TMP as compared to NaIO(3) group(P<0.01). The choroidal blood flow was significantly increased at all time points (at 30, 60 and 120 minutes after TMP instillation) as compared with corresponding controls. TMP had no effect on hypoxia-(1% O(2)), t-BHP- and H(2)O(2)-induced damage in RPE cells. 10(g/mL TMP could reverse 1 and 3mM NaN(3)-induced loss of viability of RPE by 18.5% (P<0.01) and 23% (P<0.01), respectively. 30µg/mL TMP could reverse 30 and 100mM NaIO(3) induced loss of viability of RPE by 18.1% (P<0.05) and 16.8% (P<0.01), respectively. CONCLUSION: TMP can significantly protect RPE from NaIO(3) induced degeneration in vivo and oxidative stress in vitro and can increase choroidal blood flow markedly in vivo.

12.
Pulm Pharmacol Ther ; 21(2): 259-63, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-17664077

RESUMO

The expectorant activity of naringenin was studied. Mucus secretion was evaluated in mice by measuring the tracheal output of phenol red. Mucociliary movement function was investigated using a migration method of carbon granules in unanesthetized pigeons. And the effect of naringenin on the secretion of mucin and lysozyme was performed in the rat tracheal ring explants. Naringenin could significantly increase the secretion of phenol red from mouse tracheas at the doses of 30-67 mg/kg (i.g.) (P<0.05). Naringenin, at the dose of 90 mg/kg, increased the tracheal mucociliary velocity (TMV) to 144.4% of control (P<0.01). 100 microM naringenin could enhance the basal lysozyme secretion, but had no effect on the basal mucin secretion from the rat tracheal ring explants. Treatment with naringenin at higher concentration (10 micromol/l) could inhibit the 100 ng/ml lipopolysaccharide (LPS)-induced mucin increase. These data suggest, therefore, that naringenin has the expectorant activity.


Assuntos
Expectorantes/farmacologia , Flavanonas/farmacologia , Animais , Columbidae , Relação Dose-Resposta a Droga , Expectorantes/uso terapêutico , Feminino , Flavanonas/uso terapêutico , Técnicas In Vitro , Masculino , Camundongos , Mucinas/metabolismo , Depuração Mucociliar/efeitos dos fármacos , Muramidase/metabolismo , Ratos , Traqueia/efeitos dos fármacos , Traqueia/metabolismo
13.
Am J Chin Med ; 34(5): 901-10, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-17080553

RESUMO

The aim of this study was to investigate the effects of five alkaloids, namely verticine, verticinone, imperialine, imperialine-3beta-D-glucoside, and puqietinone, purified from Bulbus Fritillariae and used as an antitussive drug in traditional Chinese medicine, on their antimuscarinic M(2) function and the cAMP level of HEK cells transfected with muscarinic M(2) receptor plasmid. By transfecting the HEK cells with the method of calcium phosphate co-precipitation and screening with G418, the cells stably expressing M(2) receptor were identified. The expression of M(2) receptor in HEK cells was confirmed by both RT-PCR and western blot. The cAMP level in the treated cells was analyzed with RIA method ((125)I-cAMP KIT). And the results suggested that the five alkaloids could significantly elevate the cAMP concentration in the HEK cells transfected with muscarinic M(2) receptor plasmid (p < 0.01).


Assuntos
Alcaloides/farmacologia , AMP Cíclico/metabolismo , Fritillaria/química , Receptor Muscarínico M2/metabolismo , Alcaloides/química , Western Blotting , Linhagem Celular , Cevanas/química , Cevanas/farmacologia , Glucosídeos/química , Glucosídeos/farmacologia , Humanos , Estrutura Molecular , Raízes de Plantas/química , Receptor Muscarínico M2/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção
14.
Eur J Pharmacol ; 551(1-3): 125-30, 2006 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-17027747

RESUMO

15 alkaloids were isolated from five Fritillariae species and 6 derivatives were synthesized. Alkaloids having anticholinergic effect on guinea-pig tracheal smooth muscle were screened out and their mechanism was further studied on the cAMP formation in Chinese hamster ovary cells stably expressing human muscarinic M2 receptor (CHO-hM2 cells) and intracellular calcium ([Ca(2+)](i)) transient in Chinese hamster ovary cells stably expressing human muscarinic M3 receptor (CHO-hM3 cells). In normal Krebs-Henseleit (KH) solution, imperialine (15), 3beta-acetylimperialine (16) and sinpeinine A (17) concentration-dependently relaxed 1 microM carbachol-induced contraction of guinea-pig tracheal rings with EC(50) of 4.19, 1.71 and 4.67 microM, respectively. In Ca(2+)-free KH solution, 10 microM 3beta-acetylimperialine (16), imperialine (15) and sinpeinnine A (17) caused 97.42%, 5.45% and 6.55% inhibition, respectively, which indicated that the three components might inhibit muscarinic receptor in different mechanism. Results of muscarinic M2 receptor-inhibited cAMP formation in CHO-hM2 cells showed that imperialine (15) and sinpeinine A (17) could potently elevate the cAMP formation whereas 3beta-acetylimperialine (16) only had weak effect on antagonism of cAMP inhibition. Furthermore, the investigations of muscarinic M3 receptor-stimulated [Ca(2+)](i) transient in CHO-hM3 cells revealed that imperialine (15) and sinpeinine A (17) could not antagonize [Ca(2+)](i) transient, but 3beta-acetylimperialine (16) significantly inhibited [Ca(2+)](i) peak elevation with an IC(50) of 5.26 microM. The functional studies suggest that the mechanism of relaxant action of imperialine (15) and sinpeinine A (17) is due to their selective inhibitory effects on muscarinic M2 receptors and the mechanism of 3beta-acetylimperialine (16) originates from its selective muscarinic M3 receptors antagonism.


Assuntos
Alcaloides/farmacologia , Fritillaria/química , Antagonistas Muscarínicos/farmacologia , Receptores Muscarínicos/efeitos dos fármacos , Traqueia/efeitos dos fármacos , Alcaloides/síntese química , Alcaloides/isolamento & purificação , Animais , Antiasmáticos/farmacologia , Atropina/farmacologia , Células CHO , Cálcio/metabolismo , Carbacol/farmacologia , Cevanas/farmacologia , Agonistas Colinérgicos/farmacologia , Cricetinae , Cricetulus , AMP Cíclico/metabolismo , Relação Dose-Resposta a Droga , Cobaias , Humanos , Antagonistas Muscarínicos/síntese química , Antagonistas Muscarínicos/isolamento & purificação , Relaxamento Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Raízes de Plantas , Receptor Muscarínico M2/efeitos dos fármacos , Receptor Muscarínico M3/efeitos dos fármacos , Receptores Muscarínicos/genética , Receptores Muscarínicos/metabolismo , Fatores de Tempo , Traqueia/metabolismo , Transfecção
15.
Planta Med ; 72(9): 814-8, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16881015

RESUMO

18 alkaloids were successfully isolated from five Fritillaria species and 5 derivatives were synthesized. Their effects on the bioactivity of human whole blood cholinesterase (ChE) were assessed. The results showed that N-demethylpuqietinone, hupeheninoside, ebeiedinone, yibeinoside A and chuanbeinone inhibited the bioactivity of human whole blood ChE at the concentration of 1.0 x 10 ( - 4) M, with the inhibitory effects of 55.5 +/- 2.7 %, 66.8 +/- 2.0 %, 69.0 +/- 1.7 %, 71.2 +/- 1.8 % and 70.7 +/- 3.3 %, respectively. The effects of the five alkaloids on human red blood cell (RBC) acetylcholinesterase (AChE) and human plasma butyrylcholinesterase (BChE) were further studied, and their IC (50) values for human RBC AChE were 6.4 +/- 0.003 microM, 16.9 +/- 0.018 microM, 5.7 +/- 0.004 microM, 6.5 +/- 0.013 microM and 7.7 +/- 0.001 microM, respectively, and the IC50 values for human plasma BChE were 12.5 +/- 0.026 microM, 2.1 +/- 0.005 microM, 5.2 +/- 0.002 microM, 7.3 +/- 0.005 microM and 0.7 +/- 0.001 microM, respectively. These data suggest, therefore, that N-demethylpuqietinone, hupeheninoside, ebeiedinone, yibeinoside A and chuanbeinone have both anti-RBC AChE and anti-plasma BChE activities, N-demethylpuqietinone is a selective inhibitor of AChE, whereas hupeheninoside and chuanbeinone are the selective inhibitors of BChE.


Assuntos
Alcaloides/farmacologia , Inibidores da Colinesterase/farmacologia , Fritillaria/química , Esteroides/farmacologia , Alcaloides/química , Células Sanguíneas/metabolismo , Cevanas/química , Cevanas/isolamento & purificação , Cevanas/farmacologia , Inibidores da Colinesterase/química , Glucosídeos/química , Glucosídeos/isolamento & purificação , Glucosídeos/farmacologia , Humanos , Esteroides/química
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