RESUMEN
Alzheimer's disease (AD) has become the most prevalent neurodegenerative disorder. Given the pathogenesis of AD is unclear, there is currently no drug approved to halt or delay the progression of AD. Therefore, it is pressing to explore new targets and drugs for AD. In China, polyphenolic Chinese herbal medicine has been used for thousands of years in clinical application, and no toxic effects have been reported. In the present study, using Dgalactose and aluminuminduced rat model, the effects of paeonol on AD were validated via the Morris water maze test, open field test, and elevated plus maze test. Neuronal morphology in frontal cortex was assessed using ImageJ's Sholl plugin and RESCONSTRUCT software. RhoA/Rock2/Limk1/cofilin1 signaling pathwayrelated molecules were determined by Western blotting. Cofilin1 and pcofilin1 were analyzed by immunofluorescence. Results showed that pretreatment with paeonol attenuated Dgalactose and aluminuminduced behavioral dysfunction and ADlike pathological alterations in the frontal cortex. Accompanied by these changes were the alterations in the dendrite and dendritic spine densities, especially the mushroomtype and filopodiatype spines in the apical dendrites, as well as actin filaments. In addition, the activity and intracellular distribution of cofilin1 and the molecules RhoA/Rock2/Limk1 that regulate the signaling pathway for cofilin1 phosphorylation have also changed. Our data suggests that paeonol may be through reducing Aß levels to alleviate the loss of fibrillar actin and dendrites and dendritic spines via the Rho/Rock2/Limk1/cofilin1 signaling pathway in the frontal cortex, and ultimately improving ADlike behavior.
Asunto(s)
Aluminio/farmacología , Enfermedad de Alzheimer/metabolismo , Espinas Dendríticas/metabolismo , Galactosa/farmacología , Proteína de Unión al GTP rhoA/metabolismo , Enfermedad de Alzheimer/patología , Animales , Espinas Dendríticas/efectos de los fármacos , Espinas Dendríticas/patología , Hipocampo/efectos de los fármacos , Quinasas Lim/efectos de los fármacos , Quinasas Lim/metabolismo , Neuronas/efectos de los fármacos , Fosforilación/efectos de los fármacos , Proteína de Unión al GTP rhoA/efectos de los fármacosRESUMEN
OBJECTIVE: To observe effects of Biejiajian Pills on hepatocarcinoma (HCC) cell vasculogenic mimicry (VM) and explore the molecular mechanism by which Biejiajian Pills inhibits HCC metastasis and invasion. METHODS: Forty male SD rats were randomly divided into 4 groups for gastric lavage of normal saline or high, moderate or low doses of Biejiajian Pills (twice daily) for 4 consecutive days. The sera were collected from the rats for treatment of cultured human HCC HepG2 cells. VM formation in the cells was detected using an image acquisition and analysis system 24 h after incubation of the cells with the sera and with the RhoA/ROCK inhibitor Y-27632(P). The expression levels of RhoA and ROCK1 in the cells were detected using Western blotting, and the contents of VE-cadherin and PI3K in the culture supernatant were determined using ELISA. RESULTS: Treatment with the sera from Biejiajian Pills-treated rats significantly inhibited formation of VM in HepG2 cells, and the diameters of VM formed were significantly greater than those in the positive control group (P < 0.01). Y-27632 completely inhibited the formation of VM in HepG2 cells (P < 0.01). Treatments with Biejiajian Pills and Y-27632 both inhibited the expression of RhoA and ROCK1 (P < 0.05) and significantly lowered the contents of VE-cadherin and PI3K in the culture supernatant (P < 0.05). CONCLUSIONS: Biejiajian Pills can inhibit the formation of VM in HCC cells in vitro possibly by inhibiting the RhoA/ROCK pathways and the expressions of VE-cadherin and PI3K.
Asunto(s)
Carcinoma Hepatocelular/irrigación sanguínea , Medicamentos Herbarios Chinos/farmacología , Neoplasias Hepáticas/irrigación sanguínea , Neovascularización Patológica/prevención & control , Quinasas Asociadas a rho/metabolismo , Proteína de Unión al GTP rhoA/metabolismo , Animales , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Línea Celular Tumoral , Células Hep G2 , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Masculino , Invasividad Neoplásica , Neovascularización Patológica/metabolismo , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Quinasas Asociadas a rho/efectos de los fármacos , Proteína de Unión al GTP rhoA/efectos de los fármacosRESUMEN
OBJECTIVES: Total flavones of Rhododendron simsii Planch flower (TFR) are an effective part extracted from the flower. The present study was designed to investigate the protective effect of TFR in isolated rat heart following global ischaemia-reperfusion and the possible underlying mechanisms. METHODS: Langendorff perfusion apparatus was used to perfuse isolated rat heart which was subjected to global ischaemia-reperfusion. The hemodynamic parameters were continuously monitored. Coronary flow as well as lactate dehydrogenase (LDH), creatine phosphokinase-MB (CK-MB) and cardiac troponin I (cTnI) in coronary effluents was measured. RhoA activity and urotensin receptor (UTR) and Rho-related coiled-coil-forming protein kinase (ROCK) protein expressions in rat myocardium were examined, respectively. Cardiac dysfunction was indicated by the alterations of hemodynamic parameters and the reduced coronary flow. KEY FINDINGS: Total flavones of Rhododendron simsii Planch flower significantly improved ischaemia-reperfusion-induced cardiac dysfunction and leakages of LDH, CK-MB and cTnI, and inhibited myocardial ischaemia-reperfusion-increased RhoA activity and UTR, ROCK1 and ROCK2 protein expressions. The improvement of TFR in the cardiac dysfunction and the leakage of LDH, CK-MB and cTnI were markedly attenuated under the UTR blockade and ROCK inhibition. TFR-inhibited RhoA activity was decreased under the UTR blockade. CONCLUSIONS: Total flavones of Rhododendron simsii Planch flower had a protective effect on ischaemia-reperfusion injury in isolated rat heart, which may be attributed to the blocking of UTR and subsequent inhibition of the RhoA-ROCK pathway.
Asunto(s)
Flavonas/farmacología , Daño por Reperfusión Miocárdica/prevención & control , Extractos Vegetales/farmacología , Rhododendron , Animales , Circulación Coronaria/efectos de los fármacos , Forma MB de la Creatina-Quinasa/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Femenino , Flavonas/administración & dosificación , Flores , L-Lactato Deshidrogenasa/efectos de los fármacos , Masculino , Extractos Vegetales/administración & dosificación , Sustancias Protectoras , Ratas , Ratas Sprague-Dawley , Receptores Acoplados a Proteínas G/efectos de los fármacos , Troponina I/efectos de los fármacos , Verapamilo/farmacología , Quinasas Asociadas a rho/efectos de los fármacos , Proteína de Unión al GTP rhoA/efectos de los fármacosRESUMEN
BACKGROUND: Omega-3 polyunsaturated fatty acids (n-3 PUFAs) show beneficial effects on cardiovascular health and cognitive functions, but the underlying molecular mechanisms are not completely understood. Because of the fact that cytoskeleton dynamics affect almost every cellular process, the regulation of cytoskeletal dynamics could be a new pathway by which n-3 PUFAs exert their effects on cellular level. METHODS: A 12-week open-label intervention study with 12 healthy men was conducted to determine the effects of 2.7 g/d n-3 PUFA on changes in mRNA expression of cytoskeleton-associated genes by quantitative real-time PCR in whole blood. Furthermore, the actin content in red blood cells was analyzed by immunofluorescence imaging. RESULTS: N-3 PUFA supplementation resulted in a significant down-regulation of cytoskeleton-associated genes, in particular three GTPases (RAC1, RHOA, CDC42), three kinases (ROCK1, PAK2, LIMK), two Wiskott-Aldrich syndrome proteins (WASL, WASF2) as well as actin related protein 2/3 complex (ARPC2, ARPC3) and cofilin (CFL1). Variability in F-actin content between subjects was high; reduced actin content was only reduced within group evaluation. CONCLUSIONS: Reduced cytoskeleton-associated gene expression after n-3 PUFA supplementation suggests that regulation of cytoskeleton dynamics might be an additional way by which n-3 PUFAs exert their cellular effects. Concerning F-actin, this analysis did not reveal unmistakable results impeding a generalized conclusion.
Asunto(s)
Citoesqueleto/efectos de los fármacos , Ácidos Grasos Omega-3/farmacología , Complejo 2-3 Proteico Relacionado con la Actina/efectos de los fármacos , Adulto , Cofilina 1/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Técnica del Anticuerpo Fluorescente , Expresión Génica/efectos de los fármacos , Humanos , Quinasas Lim/efectos de los fármacos , Masculino , Reacción en Cadena en Tiempo Real de la Polimerasa , Familia de Proteínas del Síndrome de Wiskott-Aldrich/efectos de los fármacos , Proteína Neuronal del Síndrome de Wiskott-Aldrich/efectos de los fármacos , Proteína de Unión al GTP cdc42/efectos de los fármacos , Quinasas p21 Activadas/efectos de los fármacos , Proteína de Unión al GTP rac1/efectos de los fármacos , Quinasas Asociadas a rho/efectos de los fármacos , Proteína de Unión al GTP rhoA/efectos de los fármacosRESUMEN
This study reports the antimigration, anti-invasive effect of glabridin, a flavonoid obtained from licorice, in human non-small cell lung cancer A549 cells. Glabridin exhibited effective inhibition of cell metastasis by decreasing cancer cell migration and invasion of A549 cells. In addition, glabridin also decreased A549-mediated angiogenesis. Further investigation revealed that glabridin's inhibition of cancer angiogenesis was also evident in a nude mice model. Blockade of A549 cells migration was associated with an increase of ανß3 integrin proteosome degradation. Glabridin also decreased the active forms of FAK and Src, and enhanced levels of inactivated phosphorylated Src (Tyr 527), decreasing the interaction of FAK and Src. Inhibition of the FAK/Src complex by glabridin also blocked Akt activation, resulting in reduced activation of RhoA and myosin light chain phosphorylation. This study demonstrates that glabridin may be a novel anticancer agent for the treatment of lung cancer in 3 different ways: inhibition of migration, invasion, and angiogenesis.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Proteína-Tirosina Quinasas de Adhesión Focal/efectos de los fármacos , Isoflavonas/farmacología , Neoplasias Pulmonares/metabolismo , Fenoles/farmacología , Transducción de Señal/efectos de los fármacos , Proteína de Unión al GTP rhoA/efectos de los fármacos , Animales , Carcinoma de Pulmón de Células no Pequeñas/enzimología , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Transición Epitelial-Mesenquimal/efectos de los fármacos , Proteína-Tirosina Quinasas de Adhesión Focal/metabolismo , Humanos , Integrina beta3/efectos de los fármacos , Integrina beta3/metabolismo , Neoplasias Pulmonares/enzimología , Ratones , Ratones Desnudos , Cadenas Ligeras de Miosina/metabolismo , Neovascularización Patológica/enzimología , Neovascularización Patológica/metabolismo , Fosforilación/efectos de los fármacos , Proteína de Unión al GTP rhoA/metabolismo , Familia-src Quinasas/efectos de los fármacos , Familia-src Quinasas/metabolismoRESUMEN
We have previously reported neuroprotection in spinal cord injury (SCI) by Lipitor [atorvastatin (AT)]-pre-treatment. Though informative, pre-treatment studies find only limited clinical application as trauma occurrence is unpredictable. Therefore, this study investigates the efficacy of AT treatment post-SCI. In a rat model of contusion-SCI resulting in complete hindlimb paralysis, AT treatment (5 mg/kg; gavage) was begun 2, 4, or 6 h post-SCI followed by a once daily dose thereafter for 6 weeks. While the placebo vehicle (VHC)-SCI rats showed substantial functional deficit, AT-SCI animals exhibited significant functional recovery. AT diminished injury-induced blood-spinal cord barrier (BSCB) dysfunction with significantly reduced infiltration and tumor necrosis factor-alpha/interleukin-1beta/inducible nitric oxide synthase expression at site of injury. BSCB protection in AT-SCI was attributable to attenuated matrix metalloproteinase-9 (MMP9) expression - a central player in BSCB disruption. Furthermore, endothelial MMP9 expression was found to be RhoA/ROCK pathway-mediated and regulated by AT through an isoprenoid-dependent mechanism. Attenuation of these early inflammatory events reduced secondary damage. Significant reduction in axonal degeneration, myelin degradation, gliosis, and neuronal apoptosis with resultant enhancement in tissue sparing was observed in AT-SCI compared with VHC-SCI. In summary, this novel report presenting the efficacy of post-injury AT treatment might be of critical therapeutic value as effective treatments are currently unavailable for SCI.