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1.
Int J Biol Macromol ; 259(Pt 1): 129006, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38176492

RESUMO

Conventional collagen-based hydrogels as wound dressing materials are usually lack of antibacterial activity and easily broken when encountering external forces. In this work, we developed a collagen peptide-based hydrogel as a wound dressing, which was composed of adipic acid dihydrazide functionalized collagen peptide (Col-ADH), oxidized dextran (ODex), polyvinyl alcohol (PVA) and borax via multiple-dynamic reversible bonds (acylhydrazone, amine, borate ester and hydrogen bonds). The injectable hydrogel exhibited satisfactory self-healing ability, antibacterial activity, mechanical strength, as well as good biocompatibility and biodegradability. In vivo experiments demonstrated the rapid hemostasis, accelerated cell migration, and promoted wound healing capacities of the hydrogel. These results indicate that the multifunctional collagen peptide-based hydrogel has great potentials in the field of wound dressings.


Assuntos
Hidrogéis , Prunella , Hidrogéis/farmacologia , Colágeno , Cicatrização , Antibacterianos/farmacologia , Peptídeos/farmacologia
2.
Free Radic Biol Med ; 164: 1-12, 2021 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-33388433

RESUMO

Non-alcoholic steatohepatitis (NASH) is the progressive stage of non-alcoholic fatty liver disease that may ultimately lead to cirrhosis and liver cancer, and there are few therapeutic options for its treatment. Physalin B (PB), a withanolide isolated from Physalis species (Solanaceae), exhibits a broad spectrum of biological activities, however, the potential role of PB in NASH has not been evaluated. The present study investigated the protective effects of PB against NASH and further elucidated the mechanisms of PB in hepatic autophagy and oxidative stress in vitro and in vivo. We conducted a series of experiments using methionine-choline deficient (MCD) diet induced NASH mice and cultured L02 cells. Serum markers of liver injury, morphology, and the histology of liver tissues were investigated. Western blot assays and quantitative real-time PCR were used to investigate the hepatoprotective effect of PB. PB significantly ameliorated hepatic injury, including hepatic index, transaminase activities, histology, and inflammation in MCD-induced mice. Moreover, PB markedly increased the expression of P62 and the ratio of LC3Ⅱ/Ⅰ in vitro and in vivo. Furthermore, PB promoted the interaction between endogenous KEAP1 and P62, reduced the interaction between KEAP1 and NRF2, activated the nuclear translocation of NRF2 and NRF2 target gene expression, and ultimately attenuated oxidative stress. In addition, knockdown of P62 blocked PB-mediated activation of NRF2 in L02 cells. These results clearly indicated that PB ameliorated NASH by stimulating autophagy and P62-KEAP1-NRF2 antioxidative signaling, suggesting that PB is expected to become a novel therapeutic drug for NASH.


Assuntos
Hepatopatia Gordurosa não Alcoólica , Animais , Autofagia , Proteína 1 Associada a ECH Semelhante a Kelch/genética , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Fígado/metabolismo , Metionina/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Hepatopatia Gordurosa não Alcoólica/metabolismo , Estresse Oxidativo , Secoesteroides
3.
Curr Med Sci ; 40(5): 917-930, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32980902

RESUMO

This study aimed to explore the protective effects of the traditional Chinese Medicine formula Shenkang VII recipe (SK-7) on renal fibrosis and the mechanisms. Renal fibrosis was induced by unilateral ureteral obstruction (UUO) in rats. The rats were then divided into 5 groups: control group (Sham operation), UUO model group, UUO model plus low to high doses of SK-7 (0.5, 1.0, or 2.0 g/kg/day, for 14 days) groups. The animals were sacrificed on the 7th or 14th day. Kidney tissues were collected for histopathological examinations (hematoxylin and eosin and Masson's trichrome staining). Immunohistochemistry was used to detect the expression of collagen type III (Col III), fibronectin (FN), α-smooth muscle actin (α-SMA), TIMP metallopeptidase inhibitor 2 (TIMP2), matrix metallopeptidase 2 (MMP2), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and monocyte chemotactic protein-1 (MCP-1). The TGF-ß1/Smad, NF-kB and Sonic hedgehog signaling proteins were detected by Western blotting. Our results showed that SK-7 prevented UUO-induced renal injury and accumulation of collagen fibrils. Renal fibrosis biomarkers Col III, FN, α-SMA and TIMP2 were increased in the rats after UUO and decreased by SK-7, while MMP2 was upregulated after treatment. SK-7 also suppressed the levels of TNF-α, IL-1ß and MCP-1 in UUO rats. In addition, SK-7 inhibited activation of the TGF-ß/Smad, NF-κB and sonic hedgehog signaling (SHH) pathways. Taken together, these findings suggest that SK-7 may regulate the synthesis and degradation of extracellular matrix, reduce inflammation and suppress the proliferation of fibroblasts, by blocking the TGF-ß1/Smad, NF-κB and SHH signaling pathways to exert its anti-renal fibrosis effect in UUO rats.


Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Fibrose/tratamento farmacológico , Proteínas Hedgehog/genética , Fator de Crescimento Transformador beta1/genética , Obstrução Ureteral/tratamento farmacológico , Animais , Medicamentos de Ervas Chinesas/química , Fibrose/etiologia , Fibrose/genética , Fibrose/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Rim/efeitos dos fármacos , Rim/patologia , Ratos , Transdução de Sinais/efeitos dos fármacos , Inibidor Tecidual de Metaloproteinase-2/genética , Obstrução Ureteral/complicações , Obstrução Ureteral/genética , Obstrução Ureteral/patologia
4.
Chin Med J (Engl) ; 131(3): 316-322, 2018 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-29363647

RESUMO

BACKGROUND: Much evidence has demonstrated that interleukin (IL)-33 plays an important role in rheumatoid arthritis (RA). However, there have been limited studies about soluble ST2, a receptor for IL-33, in RA. The aims of this study were to detect the levels of ST2 in the serum and synovial fluid of RA patients and to reveal the association of these levels with disease activity and the function of ST2 in RA. METHODS: A total of 56 RA patients and 38 age-matched healthy controls were enrolled in this study. Synovial fluid samples were collected from another 30 RA patients and 20 osteoarthritis patients. Serum and synovial fluid levels of ST2 were measured by ELISA. In addition, the levels of ST2 in the serum of RA patients before and after therapy were detected. The function of ST2 in RA was revealed by the results of an in vitro cell assay, where recombinant ST2 proteins were used to treat peripheral blood mononuclear cells (PBMCs) and RA synovial fibroblasts (RASFs). RESULTS: Serum-soluble ST2 levels were significantly higher in RA patients (127.14 ± 61.43 pg/ml) than those in healthy controls (78.37 ± 41.93 pg/ml, P < 0.01). Synovial fluid-soluble ST2 levels (41.90 ± 33.58 pg/ml) were much higher in RA patients than those in osteoarthritis patients (19.71 ± 16.72 pg/ml, P < 0.05). RA patients who received effective therapy for 6 months showed decreased serum-soluble ST2 levels (113.01 ± 53.90 pg/ml) compared to baseline (139.59 ± 68.36 pg/ml) (P = 0.01). RA patients with high disease activity had higher serum-soluble ST2 levels (162.02 ± 56.78 pg/ml) than those with low disease activity (94.67 ± 40.27 pg/ml, P = 0.001). Soluble ST2 did not affect IL-1ß, IL-6, IL-8, or tumor necrosis factor-α (TNF-α) expression in PBMCs from RA patients. However, soluble ST2 ameliorated the expressions of IL-33 and IL-1ß but not that of IL-6, IL-8, or TNF-α in resting RASFs. Interestingly, in the RASFs stimulated by TNF-α plus IL-1ß, soluble ST2 showed extensive suppressive effects on the expression of IL-6, IL-8, and TNF-α. CONCLUSION: Elevated levels of ST2 in the serum and synovial fluid were associated with disease activity and ameliorated IL-33 expression and IL-33-induced inflammation in RASFs, suggesting that soluble ST2 might be a potential therapeutic candidate for RA.


Assuntos
Artrite Reumatoide/tratamento farmacológico , Fibroblastos/metabolismo , Proteína 1 Semelhante a Receptor de Interleucina-1/metabolismo , Líquido Sinovial/metabolismo , Adulto , Idoso , Artrite Reumatoide/metabolismo , Artrite Reumatoide/fisiopatologia , Estudos de Casos e Controles , Células Cultivadas , Citocinas/metabolismo , Descoberta de Drogas , Feminino , Humanos , Inflamação/tratamento farmacológico , Inflamação/fisiopatologia , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Osteoartrite/metabolismo , Membrana Sinovial/metabolismo , Regulação para Cima
5.
Brain Res Bull ; 124: 295-305, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27345710

RESUMO

Taurine is an abundant amino acid in the nervous system, which has been proved to possess antioxidation, osmoregulation and membrane stabilization. Previously it has been demonstrated that taurine exerts ischemic brain injury protective effect. This study was designed to investigate whether the protective effect of taurine has the possibility to be applied to treat neonatal hypoxic-ischemic brain damage. Seven-day-old Sprague-Dawley rats were treated with left carotid artery ligation followed by exposure to 8% oxygen to generate the experimental group. The cerebral damage area was measured after taurine post-treatment with 2,3,5-triphenyltetrazolium chloride (TTC) staining, Hematoxyline-Eosin (HE) staining and Nissl staining. The activities of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), myeloperoxtidase (MPO), ATP and Lactic Acid productions were assayed with ipsilateral hemisphere homogenates. Western-blot and immunofluorescence assay were processed to detect the expressions of AIF, Cyt C, Bax, Bcl-2 in brain. We found that taurine significantly reduced brain infarct volume and ameliorated morphological injury obviously reversed the changes of SOD, MDA, GSH-Px, T-AOC, ATP, MPO, and Lactic Acid levels. Compared with hypoxic-ischemic group, it showed marked reduction of AIF, Cyt C and Bax expressions and increase of Bcl-2 after post-treatment. We conclude that taurine possesses an efficacious neuroprotective effect after cerebral hypoxic-ischemic damage in neonatal rats.


Assuntos
Lesões Encefálicas/etiologia , Lesões Encefálicas/prevenção & controle , Hipóxia-Isquemia Encefálica/complicações , Fármacos Neuroprotetores/uso terapêutico , Taurina/uso terapêutico , Animais , Animais Recém-Nascidos , Fator de Indução de Apoptose/metabolismo , Infarto Encefálico/etiologia , Infarto Encefálico/prevenção & controle , Feminino , Glutationa/metabolismo , Humanos , Ácido Láctico/metabolismo , Masculino , Malondialdeído/metabolismo , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Ratos Sprague-Dawley , Superóxido Dismutase/metabolismo , Taurina/farmacologia
6.
Zhongguo Zhong Xi Yi Jie He Za Zhi ; 36(12): 1504-1509, 2016 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-30650298

RESUMO

Objective To observe the effects of oxymatrine (OM) in inhibiting the proliferation and percentage of cancer stem cell like cell of human breast cancer MCF-7 cells, and to study its molecular mechanism. Methods MCF-7 cells were taken as subject. Side population cells (SP) of cancer stem cell like cells rich in MCF-7 cells were isolated using side population (SP) method. The proliferation properties of SP cells and non-side population (non-SP) cells were detected by MTT assay. The proliferation and intervention of cisplatin (DDP) and OM at various concentrations were detected as well. The ex- pression levels of ß-catenin gene and protein were detected using flow cytometry and immunofluorescence technique. Results (1) OM in different concentrations had various inhibitions to the proliferation of subpopulations of MCF-7 cells. Of them, it had weakest inhibition on non-SP cells, weaker inhibition on unsorted cells, and strongest inhibition on SP cells. DDP in different concentrations had strongest inhibition on non-SP cells, weaker inhibition on unsorted cells, and weakest inhibition on SP cells. (2) SP cells accounted for 3. 1%, 1. 7%, and 0. 2% of the total cells after OM acted at 0. 25, 0. 50, 1. 00 mg/mL, respectively. The expression rate of phosphorylated ß-catenin was 42. 62% ±2. 62% after SP cells were ac- ted by OM, with statistical difference as compared with the blank control group [ (22. 8% ±1. 66%) ,P < 0. 01]. (3) Compared with SP cells without OM treatment, the expression of ß-catenin in OM treated SP cells was obviously reduced. Besides, they were specifically distributed under the cytomembrane, with nuclear translocation obviously reduced. Conclusion OM could intervene biological behaviors of cancer stem cell like cell of MCF-7 cells possibly through Inhibiting the activation of Wnt/ß-catenin signaling pathway.


Assuntos
Alcaloides , Neoplasias da Mama , Quinolizinas , Via de Sinalização Wnt , Alcaloides/farmacologia , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células , Humanos , Células MCF-7 , Quinolizinas/farmacologia , Via de Sinalização Wnt/efeitos dos fármacos , beta Catenina
7.
Int J Clin Exp Pathol ; 8(9): 11236-47, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26617848

RESUMO

It is extremely difficult to discriminate between follicular thyroid carcinoma (FTC) and follicular thyroid adenoma (FTA) before surgery, because the morphologies of carcinoma cells and adenoma cells obtained by fine needle aspiration biopsy (FNAB) are similar. Molecular markers may be helpful on this issue. The purpose of this study was to assess the role of GPER1, EGFR and CXCR1 in differential diagnosis between FTC and FTA. GPER1, EGFR and CXCR1 mRNA expression levels were examined in 15 FTCs and 10 FTAs using real-time RT-PCR. FTC showed to have significantly increased mRNA levels of the three molecules compared to FTA (P < 0.001 for all the three molecules). GPER1, EGFR and CXCR1 protein expression in 106 FTCs and 128 FTAs were analyzed using immunohistochemistry. The rates of GPER1, EGFR and CXCR1 high expression were 73.6%, 72.6% and 70.8% in FTC and 30.5%, 28.1% and 27.3% in FTA, respectively. Statistical analysis showed that GPER1, EGFR and CXCR1 protein expression were correlated with one another in FTC and concomitant high expression of the three molecules had stronger correlation with the occurrence of FTC than did each alone. The positive predictive values (PPV) for concomitant high expression of the three molecules for discriminating between FTC and FTA were 91.0% for GPER1/EGFR, 93.8% for GPER1/CXCR1, 92.3% for EGFR/CXCR1 and 98.2% for GPER1/EGFR/CXCR1, respectively. These results indicated that the evaluation of GPER1, EGFR and CXCR1 concomitant high expression may be helpful in differential diagnosis between FTC and FTA.


Assuntos
Adenocarcinoma Folicular/química , Adenoma/química , Biomarcadores Tumorais/análise , Receptores ErbB/análise , Receptores de Estrogênio/análise , Receptores Acoplados a Proteínas G/análise , Receptores de Interleucina-8A/análise , Neoplasias da Glândula Tireoide/química , Adenocarcinoma Folicular/genética , Adenocarcinoma Folicular/patologia , Adenoma/genética , Adenoma/patologia , Adulto , Biomarcadores Tumorais/genética , Diagnóstico Diferencial , Receptores ErbB/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Estrogênio/genética , Receptores Acoplados a Proteínas G/genética , Receptores de Interleucina-8A/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/patologia , Regulação para Cima
8.
Int J Mol Med ; 36(3): 633-44, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26135032

RESUMO

Matrine, an active constituent of the Chinese herb, Sophora flavescens Ait., and it is known for its antioxidant, anti-inflammatory and antitumor activities. It has been demonstrated that matrine exerts protective effects against heart failure by decreasing the expression of caspase-3 and Bax, and increasing Bcl­2 levels. In this study, we aimed to determine whether these protective effects of matrine can be applied to cerebral ischemia. Following 7 successive days of treatment with matrine (7.5, 15 and 30 mg/kg) and nimodipine (1 mg/kg) by intraperitoneal injection, male Institute of Cancer Research (ICR) mice were subjected to middle cerebral artery occlusion (MCAO). Following reperfusion, the neurobehavioral score and brain infarct volume were estimated, and morphological changes were analyzed by hematoxylin and eosin (H&E) staining and electron microscopy. The percentage of apoptotic neurons was determined by flow cytometry. The levels of oxidative stress were assessed by measuring the levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), and the total antioxidant capacity (T-AOC). Western blot analysis and immunofluorescence staining were used to examine the expression of the apoptosis-related proteins, caspase-3, Bax and Bcl-2. Our results revealed that pre-treatment with matrine significantly decreased the infarct volume and improved the neurological scores. Matrine also reduced the percentage of apoptotic neurons and relieved neuronal morphological damage. Furthermore, matrine markedly decreased the MDA levels, and increased SOD, GSH-Px and CAT activity, and T-AOC. Western blot analysis and immunofluorescence staining revealed a marked decrease in caspase-3 expression and an increase in the Bcl-2/Bax ratio in the group pre-treated with matrine (30 mg/kg) as compared with the vehicle-treated group. The findings of the present study demonstrate that matrine exerts neuroprotective effects against cerebral ischemic injury and that these effects are associated with its antioxidant and anti-apoptotic properties.


Assuntos
Alcaloides/uso terapêutico , Antioxidantes/uso terapêutico , Isquemia Encefálica/tratamento farmacológico , Isquemia Encefálica/patologia , Encéfalo/efeitos dos fármacos , Fármacos Neuroprotetores/uso terapêutico , Quinolizinas/uso terapêutico , Alcaloides/isolamento & purificação , Animais , Antioxidantes/isolamento & purificação , Apoptose/efeitos dos fármacos , Encéfalo/patologia , Isquemia Encefálica/metabolismo , Masculino , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Quinolizinas/isolamento & purificação , Sophora/química , Matrinas
9.
Med Chem ; 8(4): 711-6, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22530912

RESUMO

Multidrug resistance in cancer is a major cause of failure in cancer chemotherapy. In search of new compounds with strong reversal activity and simple molecular structure, we have synthesized a series of compounds in which different substituents were linked to the 2-position of the 6,7-dimethoxy-1-(3,4-dimethoxybenzyl)- tetrahydroisoquinoline system. Compounds were analyzed for their cytotoxicity by MTT in K562 cell line in vitro, all of the derivatives exhibited little cytotoxic activity. In the meantime, these compounds were evaluated by MTT in K562/A02 cell line in vitro, 6e, 6h and 7c exhibited similar or more potent activities than verapamil with the IC50 values at 0.66, 0.65 and 0.96µM, and with the ratio factor of 24.13, 24.50 and 16.59, respectively.


Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Desenho de Fármacos , Tetra-Hidroisoquinolinas/síntese química , Tetra-Hidroisoquinolinas/farmacologia , Antineoplásicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Humanos , Concentração Inibidora 50 , Células K562 , Estrutura Molecular , Tetra-Hidroisoquinolinas/química
10.
Thromb Haemost ; 103(1): 224-33, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20062929

RESUMO

Tissue factor (TF) antagonists targeting the factor VII (FVII) binding domain have been shown to interrupt acute vascular thrombus formation without impairing haemostasis in non-human primates. In this study, we evaluate whether a human/mouse chimeric monoclonal antibody (ALT-836, formerly known as Sunol-cH36) blocking the factor X/factor IX (FX/FIX) binding site of tissue factor could achieve similar clinical benefits in an arterial thrombosis model induced by surgical endarterectomy in chimpanzees. In this model, sequential surgical endarterectomies on right and left superficial femoral arteries were performed 30 days apart in five chimpanzees. A bolus (1 mg/kg) of ALT-836 was injected intravenously immediately preceding the restoration of flow in the endarterectomised femoral artery. Pre-surgical labelling of autologous platelets using (111)In-Oxine and post-surgical gamma camera imaging of (111)In-platelet deposition at endarterectomy sites was performed. The manipulated arterial segments were harvested for patency analysis 30 days following surgery. The results indicate that ALT-836 was highly effective at reducing acute vascular thrombosis, with no significant variations in surgical blood loss and template-bleeding time in the treated group compared to the control animals. These data suggest that ALT-836 is an effective and safe antithrombotic agent in preventing TF-initiated vascular thrombogenesis without compromising haemostasis.


Assuntos
Anticorpos Monoclonais/farmacologia , Fator X/metabolismo , Fibrinolíticos/farmacologia , Tromboplastina/antagonistas & inibidores , Trombose/prevenção & controle , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/efeitos adversos , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/farmacocinética , Especificidade de Anticorpos , Sítios de Ligação , Coagulação Sanguínea/efeitos dos fármacos , Perda Sanguínea Cirúrgica/prevenção & controle , Células CHO , Cricetinae , Cricetulus , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Endarterectomia , Fator IX/metabolismo , Fator VIIa/metabolismo , Feminino , Artéria Femoral/cirurgia , Fibrinolíticos/administração & dosagem , Fibrinolíticos/efeitos adversos , Fibrinolíticos/farmacocinética , Hemorragia/induzido quimicamente , Humanos , Injeções Intravenosas , Camundongos , Camundongos Endogâmicos BALB C , Pan troglodytes , Cintilografia , Proteínas Recombinantes de Fusão/farmacologia , Tromboplastina/imunologia , Tromboplastina/metabolismo , Trombose/sangue , Trombose/diagnóstico por imagem , Trombose/etiologia
11.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 40(6): 1015-20, 2009 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-20067110

RESUMO

OBJECTIVE: To investigate the apoptosis induced by Pteris semipnnata L 5F (PsL5F) in human anaplastic thyroid carcinoma FRO cells and its molecular mechanism. METHODS: Human anaplastic thyroid carcinoma FRO cells were treated with PsL5F, and the growth inhibition rate was evaluated by MTT assay. The cell apoptosis rate was assessed by Annexin V-FITC fluorescence staining and flow cytometry. Intracellular reactive oxygen species (ROS) levels were analyzed by CM-H2DCFDA fluorescence staining and flow cytometry. Mitochondrial membrane potential (MMP) was measured by DiOD6 (3) fluorescence staining and flow cytometry. The levels of Bax, Cyto C, AIF and cleaved PARP were analyzed by Western blotting. The activity of caspase-3 was assayed by caspase-3 colorimetric assay kit. RESULTS: PsL5F has significant growth inhibitory action on FRO cells in dose and time dependent manners. Under the treatment of 100 mg/L of PsL5F, the percentage of apoptotic cells with phosphatidylserine (PS) externalization was gradually increased in time dependent manner. The rise of ROS level in FRO cells was observed as early as 1h after treated with PsL5F. The elevation of intracellular ROS levels and cell apoptosis could be inhibited by glutathione (GSH), a scavenger of ROS. The MMP in FRO cells was gradually reduced by PsL5F, and the reduction of MMP can be inhibited by GSH. Meanwhile, the levels of Bax in fraction of mitochondrial membrane, Cyto C and AIF in fraction of cytosol were gradually increased. PsL5F can cause the increase of caspase-3 activity and cleavage of PARP, a substrate of caspase-3. CONCLUSION: PsL5F can inhibit growth of human anaplastic thyroid carcinoma FRO cells through inducing apoptosis. The rise of ROS levels in FRO cells plays important role as a secondary messenger in apoptosis induced by PsL5F. Mitochondrium is an important target of PsL5F. Cell apoptosis induced by PsL5F in FRO cells was carried out through translocation of Bax to mitochondrial membrane, reduction of MMP, release of Cyto C and AIF from mitochondria to cytosol, and activation of caspases cascade reaction.


Assuntos
Apoptose/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Pteris/química , Linhagem Celular Tumoral , Diterpenos/isolamento & purificação , Diterpenos/farmacologia , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Carcinoma Anaplásico da Tireoide , Neoplasias da Glândula Tireoide/patologia
12.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 39(6): 925-8, 2008 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-19253827

RESUMO

OBJECTIVE: To study the effects of mPer1 gene on the response of mammary carcinoma EMT6 cells to Adriamycin in vitro. METHODS: The eukaryotic expression vector pcDNA3. 1 (+)-mPer1 was transfected into the EMT6 cells (EMT6-mPerl). The vector pcDNA3. 1(+) transfect was also performed to serve as the control (EMT6-vect). The transfect efficiency was detected by RT-PCR and Western Blotting. The transfect cells were treated with Adriamycin in vitro. The apoptosis and distribution of cells in the cell cycle were analysed by FCM. The cell proliferation was detected by MTT assay. RT-PCR was used to show the mRNA expression of apoptosis-related genes. RESULTS: The mPerl-transfected EMT6 cells revealed S phase arrest, increased rate of apoptosis [EMT6-vect: (65.65 +/- 0.07)%; EMT6-mPer1: (72.35 +/- 0.57)%], decreased cell proliferation CEMT6-vect: (42.18 +/- 5.73)%; EMT6-mPer1: (53.28 +/- 7.32%)%] and stronger expression of p53 mRNA in RT-PCR (EMT6-vect, 0.48 +/- 0.08; EMT6-mPer1: 1.18 +/- 0.02). CONCLUSION: mPer1 gene can improve the drug sensitivity of this cell line to ADM in vitro.


Assuntos
Apoptose/efeitos dos fármacos , Doxorrubicina/farmacologia , Neoplasias Mamárias Experimentais/genética , Proteínas Circadianas Period/genética , Transfecção , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sinergismo Farmacológico , Vetores Genéticos , Neoplasias Mamárias Experimentais/patologia , Camundongos , Proteínas Circadianas Period/metabolismo , Proteínas Circadianas Period/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo
13.
Zhongguo Zhong Yao Za Zhi ; 32(7): 609-12, 2007 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-17583203

RESUMO

OBJECTIVE: To discuss the effect of Yisui Shengxue granules on expression of alpha-hemoglobin stabilizing protein (AHSP) mRNA in different developmental stages mice. METHOD: The total RNAs were extracted from the bone marrow karyocyte of normal adult mice and the karyocyte of fetus liver and fetus spleen in pregnanted mice (pregnanted 21 days) and fetal mice (pregnanted 14 days). The expression level of AHSP mRNA in different developmental stages mice interfered with Yisui Shengxue granules was measured by real-time PCR. RESULT: The intervention of Yisui Shengxue granules could significantly up-regulated the expression levels of AHSP mRNA in normal adult mice. CONCLUSION: The result revealed that one of possible molecular mechanism of the effects caused by Yisui Shengxue granules is that it can promote the AHSP gene expression, reduce the free a-globin deposit, then prevent the poison to erythrocyte and decrease the haemolysis.


Assuntos
Proteínas Sanguíneas/genética , Medicamentos de Ervas Chinesas/farmacologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Chaperonas Moleculares/genética , Plantas Medicinais/química , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Combinação de Medicamentos , Medicamentos de Ervas Chinesas/isolamento & purificação , Eritrócitos/citologia , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Feminino , Fígado/citologia , Fígado/embriologia , Fígado/metabolismo , Masculino , Camundongos , Gravidez , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Distribuição Aleatória , Baço/citologia , Baço/embriologia , Baço/metabolismo , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genética
14.
Luminescence ; 18(4): 218-23, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-12950058

RESUMO

Studies were performed to compare green fluorescent protein (GFP)-transfected and fi re fl y luciferase (Luc)-transfected MCF-7 human breast tumour cells both in vitro and in vivo. For in vitro studies, cells were serially diluted in 96-well microplates and analysed using a NightOwl LB 981 Molecular Light Imager and a Victor multilabel reader. For in vivo studies, nude mice were injected either intraperitoneally, intravenously or subcutaneously with transfected cells and then imaged using the NightOwl Imager after intraperitoneal injection of d-luciferin for Luc tumours, or excitation at 470 nm for GFP tumours. In vitro imaging studies revealed that both GFP and Luc transfectants were quantifiable. However, the Luc-transfected cells were detectable at a significantly lower concentration compared to GFP transfectants. In vivo studies demonstrated that GFP-transfected tumours were detectable as subcutaneous and intraperitoneal tumours but not as deep tissue lesions, whereas Luc-transfected tumours were detectable as subcutaneous and intraperitoneal tumours and as deep tissue lesions resulting from intraperitoneal or intravenous inoculation. These findings demonstrate that GFP-transfected cells may be useful for imaging studies of superficial tumours where both excitation and emission wavelengths are able to penetrate tissues, whereas luciferase-transfected cells appear superior for imaging studies of primary and metastatic tumours in distant sites and deep tissues.


Assuntos
Diagnóstico por Imagem/métodos , Luciferases , Proteínas Luminescentes , Neoplasias Experimentais/patologia , Animais , Linhagem Celular Tumoral , Feminino , Genes Reporter , Proteínas de Fluorescência Verde , Humanos , Luciferases/genética , Proteínas Luminescentes/genética , Camundongos , Camundongos Nus , Neoplasias Experimentais/diagnóstico , Transfecção , Transplante Heterólogo
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