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1.
Nutrients ; 13(7)2021 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-34371956

RESUMO

We examined the immunomodulatory and anti-inflammatory effects of asiatic acid (AA) in atopic dermatitis (AD). AA treatment (5-20 µg/mL) dose-dependently suppressed the tumor necrosis factor (TNF)-α level and interleukin (IL)-6 protein expression in interferon (IFN)-γ + TNF-α-treated HaCaT cells. The 2,4-dinitrocholrlbenzene (DNCB)-induced AD animal model was developed by administering two AA concentrations (30 and 75 mg/kg/d: AD + AA-L and AD + AA-H groups, respectively) for 18 days. Interestingly, AA treatment decreased AD skin lesions formation and affected other AD characteristics, such as increased ear thickness, lymph node and spleen size, dermal and epidermal thickness, collagen deposition, and mast cell infiltration in dorsal skin. In addition, in the DNCB-induced AD animal model, AA treatment downregulated the mRNA expression level of AD-related cytokines, such as Th1- (TNF-α and IL-1ß and -12) and Th2 (IL-4, -5, -6, -13, and -31)-related cytokines as well as that of cyclooxygenase-2 and CXCL9. Moreover, in the AA treatment group, the protein level of inflammatory cytokines, including COX-2, IL-6, TNF-α, and IL-8, as well as the NF-κB and MAPK signaling pathways, were decreased. Overall, our study confirmed that AA administration inhibited AD skin lesion formation via enhancing immunomodulation and inhibiting inflammation. Thus, AA can be used as palliative medication for regulating AD symptoms.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Citocinas/metabolismo , Dermatite Atópica/tratamento farmacológico , Fatores Imunológicos/farmacologia , Triterpenos Pentacíclicos/farmacologia , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/uso terapêutico , Linhagem Celular , Sobrevivência Celular , Colágeno/análise , Citocinas/genética , Dermatite Atópica/imunologia , Dermatite Atópica/patologia , Derme/patologia , Dinitroclorobenzeno , Modelos Animais de Doenças , Epiderme/patologia , Feminino , Regulação da Expressão Gênica , Humanos , Fatores Imunológicos/administração & dosagem , Fatores Imunológicos/uso terapêutico , Imunomodulação , Tecido Linfoide/patologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Mastócitos , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Triterpenos Pentacíclicos/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos
2.
Int J Mol Sci ; 22(13)2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34209790

RESUMO

We developed a multi-channel cell chip containing a three-dimensional (3D) scaffold for horizontal co-culture and drug toxicity screening in multi-organ culture (human glioblastoma, cervical cancer, normal liver cells, and normal lung cells). The polydimethylsiloxane (PDMS) multi-channel cell chip (PMCCC) was based on fused deposition modeling (FDM) technology. The architecture of the PMCCC was an open-type cell chip and did not require a pump or syringe. We investigated cell proliferation and cytotoxicity by conducting 3-(4,5-dimethylthiazol-2-yl)-2,5-dphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays and analysis of oleanolic acid (OA)-treated multi-channel cell chips. The results of the MTT and LDH assays showed that OA treatment in the multi-channel cell chip of four cell lines enhanced chemoresistance of cells compared with that in the 2D culture. Furthermore, we demonstrated the feasibility of the application of our multi-channel cell chip in various analysis methods through Annexin V-fluorescein isothiocyanate/propidium iodide staining, which is not used for conventional cell chips. Taken together, the results demonstrated that the PMCCC may be used as a new 3D platform because it enables simultaneous drug screening in multiple cells by single point injection and allows analysis of various biological processes.


Assuntos
Técnicas de Cultura de Células , Avaliação Pré-Clínica de Medicamentos , Técnicas de Cultura de Células/instrumentação , Técnicas de Cultura de Células/métodos , Linhagem Celular Tumoral , Proliferação de Células , Tamanho Celular , Técnicas de Cocultura/instrumentação , Técnicas de Cocultura/métodos , Avaliação Pré-Clínica de Medicamentos/instrumentação , Avaliação Pré-Clínica de Medicamentos/métodos , Células HeLa , Humanos , Dispositivos Lab-On-A-Chip , Teste de Materiais , Tecidos Suporte/química , Testes de Toxicidade/instrumentação , Testes de Toxicidade/métodos
3.
Nutrients ; 12(7)2020 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-32630655

RESUMO

Yeonsan Ogye is a traditional Korean chicken breed (Gallus domesticus, GD), with a dominant gene for fibromelanosis, showing entirely black fluffy head feathers, ear lobes, and pupils. GD collagen extract (78.6 g per 100 g total protein) was derived from the flesh of Yeonsan Ogye. The effects of GD collagen on bone mass, microarchitecture, osteogenic, osteoclastogenic differentiations, and function factor expression were investigated in ovariectomized (OVX) rats. GD collagen stimulated osteogenesis in OVX rats and increased tibial bone strength and calcium content. Micro-computed tomography analysis of tibia cross-sections revealed that GD collagen attenuated the OVX-induced changes in trabecular thickness, spacing, and number. GD collagen stimulated alkaline phosphatase activity, bone-specific matrix proteins (alkaline phosphatase (ALP), osteocalcin, collagen type I (COL-I)) and mineralization by activating bone morphogenetic protein 2 (BMP-2)/mothers against decapentaplegic homolog 5 (SMAD5)/runt-related transcription factor 2 (Runx2). GD collagen inhibited osteoclast differentiation and function gene markers (TRAP, cathepsin K) by interfering with the Wnt signaling, increasing OPG production, and reducing the expression of RANKL, TRAP, and cathepsin K. GD collagen promoted osteogenesis by activating the p38 signal pathway and prevented osteoclastogenesis by lowering the RANKL/OPG ratio and blocking the JNK signaling pathway. Dietary supplementation with GD collagen might inhibit osteoclastogenesis, stimulate osteoblastogenesis, and regulate bone metabolism.


Assuntos
Osso e Ossos/efeitos dos fármacos , Galinhas/metabolismo , Colágeno/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Osteoprotegerina/análise , Ligante RANK/análise , Animais , Osso e Ossos/anatomia & histologia , Osso e Ossos/fisiologia , Calcificação Fisiológica/efeitos dos fármacos , Calcificação Fisiológica/genética , Cálcio/análise , Diferenciação Celular , Linhagem Celular , Galinhas/genética , Colágeno/isolamento & purificação , Estrogênios/deficiência , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Camundongos , Osteoblastos/efeitos dos fármacos , Osteoblastos/fisiologia , Osteoclastos/efeitos dos fármacos , Osteoclastos/fisiologia , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Ovariectomia , Células RAW 264.7 , Ratos , Ratos Wistar
4.
Nanomaterials (Basel) ; 10(3)2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32183472

RESUMO

Biocompatibility is very important for cell growth using 3D printers, but biocompatibility materials are very expensive. In this study, we investigated the possibility of cell culture by the surface modification of relatively low-cost industrial materials and an efficient three-dimensional (3D) scaffold made with an industrial ABS filament for cell proliferation, spheroid formation, and drug screening applications. We evaluated the adequate structure among two-layer square shape 3D scaffolds printed by fused deposition modeling with variable infill densities (10-50%). Based on the effects of these scaffolds on cell proliferation and spheroid formation, we conducted experiments using the industrial ABS 3D scaffold (IA3D) with 40% of infill density, which presented an external dimension of (XYZ) 7650 µm × 7647 µm × 210 µm, 29.8% porosity, and 225 homogenous micropores (251.6 µm × 245.9 µm × 210 µm). In the IA3D, spheroids of cancer HepG2 cells and keratinocytes HaCaT cells appeared after 2 and 3 days of culture, respectively, whereas no spheroids were formed in 2D culture. A gold nanoparticle-coated industrial ABS 3D scaffold (GIA3D) exhibited enhanced biocompatible properties including increased spheroid formation by HepG2 cells compared to IA3D (1.3-fold) and 2D (38-fold) cultures. Furthermore, the cancer cells exhibited increased resistance to drug treatments in GIA3D, with cell viabilities of 122.9% in industrial GIA3D, 40.2% in IA3D, and 55.2% in 2D cultures when treated with 100 µM of mitoxantrone. Our results show that the newly engineered IA3D is an innovative 3D scaffold with upgraded properties for cell proliferation, spheroid formation, and drug-screening applications.

5.
Nanomaterials (Basel) ; 10(2)2020 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-32013042

RESUMO

Calcium-type montmorillonite, a phyllosilicate mineral, has diverse health benefits when introduced into the gastrointestinal tract or applied to the skin. However, the predominant use of this layered material has thus far been in traditional industries, despite its potential application in the pharmaceutical industry. We investigated the effects and mechanism of nano-montmorillonite (NM) on osteoblast and osteoclast differentiation in vivo and in vitro. We examined the osteogenic effects of NM with high calcium content (3.66 wt%) on alkaline phosphatase (ALP) activity, mineralization, bone microarchitecture, and expression level of osteoblast and osteoclast related genes in Ca-deficient ovariectomized (OVX) rats. Micro-computed tomography of OVX rats revealed that NM attenuated the low-Ca-associated changes in trabecular and cortical bone mineral density. It improved ALP activity and mineralization, as well as the expression of osteoblast and osteoclast differentiation associated genes. NM also activated the expression of runt-related transcription factor 2, osteocalcin, bone morphogenetic protein 2, and type 1 collagen via phosphorylated small mothers against decapentaplegic homolog 1/5/8 signaling. Further, NM repressed the expression of receptor activator for cathepsin K, nuclear factor kappa-B ligand and tartrate-resistant acid phosphatase. Therefore, NM inhibits osteoclastogenesis, stimulates osteoblastogenesis, and alleviates osteoporosis.

6.
Nutrients ; 11(3)2019 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-30818817

RESUMO

This study evaluated the effects of vitamin C on osteogenic differentiation and osteoclast formation, and the effects of vitamin C concentration on bone microstructure in ovariectomized (OVX) Wistar rats. Micro-computed tomography analysis revealed the recovery of bone mineral density and bone separation in OVX rats treated with vitamin C. Histomorphometrical analysis revealed improvements in the number of osteoblasts, osteoclasts, and osteocytes; the osteoblast and osteoclast surface per bone surface; and bone volume in vitamin C-treated OVX rats. The vitamin C-treated group additionally displayed an increase in the expression of osteoblast differentiation genes, including bone morphogenetic protein-2, small mothers against decapentaplegic 1/5/8, runt-related transcription factor 2, osteocalcin, and type I collagen. Vitamin C reduced the expression of osteoclast differentiation genes, such as receptor activator of nuclear factor kappa-B, receptor activator of nuclear factor kappa-B ligand, tartrate-resistant acid phosphatase, and cathepsin K. This study is the first to show that vitamin C can inhibit osteoporosis by promoting osteoblast formation and blocking osteoclastogenesis through the activation of wingless-type MMTV integration site family/ß-catenin/activating transcription factor 4 signaling, which is achieved through the serine/threonine kinase and mitogen-activated protein kinase signaling pathways. Therefore, our results suggest that vitamin C improves bone regeneration.


Assuntos
Fator 4 Ativador da Transcrição/metabolismo , Ácido Ascórbico/farmacologia , Osteoblastos/efeitos dos fármacos , Osteoclastos/efeitos dos fármacos , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Fator 4 Ativador da Transcrição/genética , Ração Animal , Animais , Densidade Óssea , Dieta , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Osteoporose/prevenção & controle , Ovariectomia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Proteínas Wnt/genética , beta Catenina/genética
7.
Molecules ; 23(11)2018 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-30413118

RESUMO

Parkinson's disease (PD), a common adult-onset neurodegenerative disorder with complex pathological mechanisms, is characterized by the degeneration of dopaminergic nigrostriatal neurons. The present study demonstrated that the herbal medicines Hepad 1 and 2 protected against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced dopaminergic neurotoxicity in C57BL/6 mice and SH-SY5Y cells. Hepad 1 and 2 remarkably alleviated the enhanced expression of pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2, macrophage-1, and phosphorylated iκB-α) and apoptotic signals (Bcl-2-associated X protein, caspase-3, and poly [ADP-ribose] polymerase-1). Additionally, Hepad reduced MPTP-induced oxidative damage by increasing the expression of anti-oxidant defense enzymes (superoxide dismutase and glutathione S-transferase) and downregulating the levels of nicotinamide adenine dinucleotide phosphate oxidase 4. This study also showed that the neuroprotective effects of Hepad include anti-inflammatory, anti-apoptotic, and anti-oxidative properties, in addition to activation of the protein kinase B, extracellular-signal-regulated kinase, and c-Jun N-terminal kinase signaling pathways. Furthermore, oral administration of Hepad 1 and 2 attenuated the death of tyrosine hydroxylase-positive substantia nigra neurons that was induced by 20 mg/kg MPTP. Therefore, our results suggest that Hepad 1 and 2 are useful for treating PD and other disorders associated with neuro-inflammatory, neuro-apoptotic, and neuro-oxidative damage.


Assuntos
1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/efeitos adversos , Fármacos Neuroprotetores/administração & dosagem , Transtornos Parkinsonianos/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Plantas Medicinais/química , Administração Oral , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Citocinas/metabolismo , Modelos Animais de Doenças , Medicina Herbária , Humanos , Camundongos , Neurônios/citologia , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Transtornos Parkinsonianos/induzido quimicamente , Transtornos Parkinsonianos/metabolismo , Extratos Vegetais/química , Extratos Vegetais/farmacologia
8.
Oncotarget ; 9(41): 26370-26386, 2018 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-29899865

RESUMO

We evaluated oleanolic acid (OA)-induced anti-cancer activity, apoptotic mechanism, cell cycle status, and MAPK kinase signaling in DU145 (prostate cancer), MCF-7 (breast cancer), U87 (human glioblastoma), normal murine liver cell (BNL CL.2) and human foreskin fibroblast cell lines (Hs 68). The IC50 values for OA-induced cytotoxicity were 112.57 in DU145, 132.29 in MCF-7, and 163.60 in U87 cells, respectively. OA did not exhibit toxicity in BNL CL. 2 and Hs 68 cell lines in our experiments. OA, at 100 µg/mL, increased the number of apoptotic cells to 27.0% in DU145, 27.0% in MCF-7, and 15.7% in U87, when compared to control cells. This enhanced apoptosis was due to increases in p53, cytochrome c, Bax, PARP-1 and caspase-3 expression in DU145, MCF-7 and U87 cell lines. OA-treated DU145 cells were arrested in G2 because of the activation of p-AKT, p-JNK, p21 and p27, and the decrease in p-ERK, cyclin B1 and CDK2 expression; OA-treated MCF-7 cells were arrested in G1 owing to the activation of p-JNK, p-ERK, p21, and p27, and the decrease in p-AKT, cyclin D1, CDK4, cyclin E, and CDK2; and OA-treated U87 cells also exhibited G1 phase arrest caused by the increase in p-ERK, p-JNK, p-AKT, p21, and p27, and the decrease in cyclin D1, CDK4, cyclin E and CDK2. Thus, OA arrested the cell cycle at different phases and induced apoptosis in cancer cells. These results suggested that OA possibly altered the expression of the cell cycle regulatory proteins differently in varying types of cancer.

9.
J Nanosci Nanotechnol ; 18(10): 6726-6731, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-29954487

RESUMO

Osteoporosis, a major public health problem, is characterised by a reduction in and deterioration of bone tissue microarchitecture, with a consequent increase in bone frailty and susceptibility to fractures. We performed ex vivo micro-computed tomography (micro-CT) and atomic-force microscopy (AFM) between normal and surgically ovariectomized animals (OVX). The tibial architecture was investigated using maximum intensity projection images. Micro-CT analysis of the tibia metaphysis was used to calculate the trabecular spacing (Tb.Sp). Following OVX surgery, the rats exhibited increased Tb.Sp. Between the bundles, there are holes in the fabric with a diameter in the range of 3 to 5 µm in normal tibia. The diameter of hole in the osteoporosis tibia is the range of 5 to 10 µm, and its area increased than normal tibia. AFM can be used for the inspection of bone surfaces and offers the possibility of quantification of local geometries, such as pit size, collagen fibril diameters, or collagen ultrastructure. Our results suggest that AFM will surely become one of the most important tools in the realm of medical research for imaging such as bone and tissues.


Assuntos
Osteoporose/diagnóstico por imagem , Tíbia/diagnóstico por imagem , Animais , Modelos Animais de Doenças , Feminino , Microscopia de Força Atômica , Osteoporose/patologia , Ovariectomia , Ratos , Tíbia/patologia , Microtomografia por Raio-X
10.
Nutr Res Pract ; 11(3): 190-197, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28584575

RESUMO

BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of 50-250 µg/mL. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to 250 µg/mL and were 149% and 129% at 250 µg/mL concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of 500 µg/mL. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

11.
Nutrients ; 9(5)2017 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-28513557

RESUMO

The present study evaluated the effects of a calcium (Ca) supplement derived from Gallus gallus domesticus (GD) on breaking force, microarchitecture, osteogenic differentiation and osteoclast differentiation factor expression in vivo in Ca-deficient ovariectomized (OVX) rats. One percent of Ca supplement significantly improved Ca content and bone strength of the tibia. In micro-computed tomography analysis, 1% Ca supplement attenuated OVX- and low Ca-associated changes in bone mineral density, trabecular thickness, spacing and number. Moreover, 1% Ca-supplemented diet increased the expression of osteoblast differentiation marker genes, such as bone morphogenetic protein-2, Wnt3a, small mothers against decapentaplegic 1/5/8, runt-related transcription factor 2, osteocalcin and collagenase-1, while it decreased the expression of osteoclast differentiation genes, such as thrombospondin-related anonymous protein, cathepsin K and receptor activator of nuclear factor kappa B. Furthermore, 1% Ca-supplemented diet increased the levels of phosphorylated extracellular signal-regulated kinase and c-Jun N-terminal kinase. The increased expression of osteoblast differentiation marker genes and activation of mitogen-activated protein kinase signaling were associated with significant increases in trabecular bone volume, which plays an important role in the overall skeletal strength. Our results demonstrated that 1% Ca supplement inhibited osteoclastogenesis, stimulated osteoblastogenesis and restored bone loss in OVX rats.


Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Osso e Ossos/química , Cálcio/administração & dosagem , Galinhas , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Osteoporose/prevenção & controle , Animais , Densidade Óssea/efeitos dos fármacos , Proteína Morfogenética Óssea 2/genética , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Osteoporose/metabolismo , Ovariectomia , Ratos , Ratos Wistar , Receptor Ativador de Fator Nuclear kappa-B/genética , Receptor Ativador de Fator Nuclear kappa-B/metabolismo , Proteína Smad5/genética , Proteína Smad5/metabolismo , Fosfatase Ácida Resistente a Tartarato/genética , Fosfatase Ácida Resistente a Tartarato/metabolismo
12.
Nutrients ; 9(1)2017 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-28067819

RESUMO

In this study, we determined the effects of hederagenin isolated from Akebia quinata fruit on alcohol-induced hepatotoxicity in rats. Specifically, we investigated the hepatoprotective, anti-inflammatory, and anti-apoptotic effects of hederagenin, as well as the role of AKT and mitogen-activated protein kinase (MAPK) signaling pathways in ethanol-induced liver injury. Experimental animals were randomly divided into three groups: normal (sham), 25% ethanol, and 25% ethanol + hederagenin (50 mg/kg/day). Each group was orally administered the respective treatments once per day for 21 days. Acetaldehyde dehydrogenase-2 mRNA expression was higher and alcohol dehydrogenase mRNA expression was lower in the ethanol + hederagenin group than those in the ethanol group. Pro-inflammatory cytokines, including TNF-α, IL-6, and cyclooxygenase-2, significantly increased in the ethanol group, but these increases were attenuated by hederagenin. Moreover, Western blot analysis showed increased expression of the apoptosis-associated protein, Bcl-2, and decreased expression of Bax and p53 after treatment with hederagenin. Hederagenin treatment attenuated ethanol-induced increases in activated p38 MAPK and increased the levels of phosphorylated AKT and ERK. Hederagenin alleviated ethanol-induced liver damage through anti-inflammatory and anti-apoptotic activities. These results suggest that hederagenin is a potential candidate for preventing alcoholic liver injury.


Assuntos
Anti-Inflamatórios/farmacologia , Apoptose/efeitos dos fármacos , Suplementos Nutricionais , Etanol/toxicidade , Inflamação/tratamento farmacológico , Ácido Oleanólico/análogos & derivados , Alanina Transaminase/sangue , Álcool Desidrogenase/genética , Álcool Desidrogenase/metabolismo , Aldeído-Desidrogenase Mitocondrial/genética , Aldeído-Desidrogenase Mitocondrial/metabolismo , Animais , Aspartato Aminotransferases/sangue , Colesterol/sangue , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Hepatopatias/tratamento farmacológico , Masculino , Ácido Oleanólico/farmacologia , Fosforilação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais , Triglicerídeos/sangue , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
13.
J Biomed Nanotechnol ; 12(2): 357-65, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27305769

RESUMO

Nanotechnology-based bio-barcode amplification analysis offers an innovative approach for detecting neurotransmitters. We evaluated the efficacy of this method for detecting norepinephrine in normal and oxidative-stress damaged dopaminergic cells. Our approach use a combination of DNA barcodes and bead-based immunoassays for detecting neurotransmitters with surface-enhanced Raman spectroscopy (SERS), and provides polymerase chain reaction (PCR)-like sensitivity. This method relies on magnetic Dynabeads containing antibodies and nanoparticles that are loaded both with DNA barcords and with antibodies that can sandwich the target protein captured by the Dynabead-bound antibodies. The aggregate sandwich structures are magnetically separated from the solution and treated to remove the conjugated barcode DNA. The DNA barcodes are then identified by SERS and PCR analysis. The concentration of norepinephrine in dopaminergic cells can be readily detected using the bio-barcode assay, which is a rapid, high-throughput screening tool for detecting neurotransmitters.


Assuntos
Ouro/química , Nanopartículas Metálicas/química , Norepinefrina/análise , Técnicas de Amplificação de Ácido Nucleico/métodos , Anexina A5/metabolismo , Apoptose , Linhagem Celular Tumoral , Fluoresceína-5-Isotiocianato/metabolismo , Humanos , Nanopartículas Metálicas/ultraestrutura , Sondas Moleculares/síntese química , Sondas Moleculares/química , Propídio/metabolismo , Reprodutibilidade dos Testes , Análise Espectral Raman
14.
J Biomed Nanotechnol ; 12(12): 2125-38, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29368908

RESUMO

In this study, we developed a novel three-dimensional (3D) cancer cell chip using a three-floor hierarchical 3D pyramid structure (3D pyramid) to simulate 3D tumor cell growth in vitro and to detect anticancer drugs. The proposed 3D pyramidbased cancer cell chip offered substantial advantages for the agglomerate formation of tumor cells, in which cells could be maintained as tumor spheroids for up to 3 weeks. Soon after HeLa tumor cells adhered to the micropatterned pillar sidewalls, they were suspended between the pillars based on scanning electron microscopy images. Treatment with the anticancer drug oleanolic acid resulted in 46.33% and 5.86% apoptotic cells on the 2D plate and 3D pyramid-based cell chip, respectively, compared with only 0.06% apoptotic cells in the control. The increase in chemoresistance to anticancer drugs in the 3D pyramid-based cell chip might be a result of cell confluence and hypoxia due to the spheroid formation of tumor cells in the 3D pyramid structure. These results indicated that the proposed cell chip could potentially be used for anticancer drug screening or can be incorporated into other models aimed at prolonging various cell functions in culture.


Assuntos
Antineoplásicos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais/instrumentação , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Análise Serial de Tecidos/instrumentação , Análise Serial de Tecidos/métodos , Apoptose/efeitos dos fármacos , Desenho de Equipamento , Células HeLa , Humanos
15.
J Microbiol Biotechnol ; 25(6): 795-802, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25639718

RESUMO

This study investigates the bioactivity of tannin from amaranth (Amaranthus caudatus L.) extracts. The antioxidant activities of the extracts from amaranth leaves, flowers, and seeds were evaluated. Tannin from leaves of amaranth has been evaluated for superoxide scavenging activity by using DPPH and ABTS(+) analysis, reducing power, protective effect against H2O2-induced oxidative damage in L-132 and BNL-CL2 cells, and inhibition of superoxide radical effects on HL-60 cells. At a concentration of 100 µg/ml, tannin showed protective effects and restored cell survival to 69.2% and 41.8% for L-132 and BNL-CL2 cells, respectively. Furthermore, at the same concentration, tannin inhibited 41% of the activity of the superoxide radical on HL-60 cells and 43.4% of the increase in nitric oxide levels in RAW 264.7 cells. The expression levels of the antioxidant-associated protein SOD-1 were significantly increased in a concentration-dependent manner in RAW 264.7 cells treated with tannin from amaranth leaves. These results suggest that tannin from the leaves of Amaranthus caudatus L. is a promising source of antioxidant component that can be used as a food preservative or nutraceutical.


Assuntos
Amaranthus/química , Sequestradores de Radicais Livres/isolamento & purificação , Sequestradores de Radicais Livres/metabolismo , Taninos/isolamento & purificação , Taninos/metabolismo , Antioxidantes/isolamento & purificação , Antioxidantes/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Flores/química , Humanos , Oxidantes/toxicidade , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/metabolismo , Folhas de Planta/química , Sementes/química
16.
Biosens Bioelectron ; 67: 739-46, 2015 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-25465795

RESUMO

Dopamine is a potent neuromodulator in the brain that influences a variety of motivated behaviors and is involved in several neurologic diseases. We evaluated a bio-barcode amplification assay for its ability to detect dopamine in a mouse model with and without prior administration of the neurotoxin 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). Our approach uses a combination of DNA barcodes and bead-based immunoassays for detecting neurotransmitters with surface-enhanced Raman spectroscopy (SERS). This method relies on a gold nanoplate with adsorbed antibodies and gold nanoparticles that are encoded with DNA and antibodies that can sandwich the target protein captured by the nanoparticle-bound antibodies. C57BL/6 mice were infused intranasally with MPTP (25mg/kg/day) over 7 consecutive days. At 7 and 21 days after the last administration of MPTP, dopamine was found by western blot analysis to have decreased in the midbrain by 37.44% and 92.95%, respectively. Furthermore, the Raman intensity of dopamine in the midbrains of MPTP-treated mice decreased by 56.77% and 61.12% on days 7 and 21, respectively. Our results demonstrate that the concentration of dopamine in midbrain and striatum of MPTP-treated mice can be easily detected using the bio-barcode assay, which is a rapid, high-throughput screening tool for detecting neurotransmitters.


Assuntos
Técnicas Biossensoriais , Dopamina/isolamento & purificação , Doença de Parkinson/diagnóstico , Transtornos Parkinsonianos/diagnóstico , Animais , Corpo Estriado , DNA/química , DNA/genética , Modelos Animais de Doenças , Dopamina/genética , Ouro/química , Humanos , Camundongos , Técnicas de Amplificação de Ácido Nucleico , Doença de Parkinson/genética , Transtornos Parkinsonianos/genética , Análise Espectral Raman
17.
J Biomed Nanotechnol ; 9(6): 1071-5, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23858972

RESUMO

In this study, the anti-tumor activity of mitoxantrone loaded on magnetic nanoparticles (MTMP) was examined using DU145 prostate cancer cells. Composite nanoparticles with an average size of 20 nm were prepared using a chemical co-precipitation technique. The MTMP nanoparticles were cytotoxic to DU145 cells and inhibited cell proliferation. The expression levels of apoptosis related proteins in DU145 cells, including PARP and caspase 3, were increased after MTMP treatment. In this study, the therapeutic potential of MTMP in targeted-therapy against prostate cancer was demonstrated and MTMP was more effective when coupled to drug delivery vehicle than pure mitoxantrone.


Assuntos
Nanopartículas de Magnetita/administração & dosagem , Mitoxantrona/administração & dosagem , Nanocápsulas/administração & dosagem , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/patologia , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Quimioterapia Combinada , Humanos , Nanopartículas de Magnetita/química , Nanopartículas de Magnetita/ultraestrutura , Masculino , Mitoxantrona/química , Nanocápsulas/química , Resultado do Tratamento
18.
J Nanosci Nanotechnol ; 12(2): 1611-5, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22630012

RESUMO

This study examined the optical characteristics of bicalutamide-loaded magnetic/ethylene glycol composite nanoparticles (BMP), as well as their anti-cancer activity against cancer cells. The gamma-Fe2O3 magnetic nanoparticles (MNPs), approximately 20 nm in diameter, were prepared via a chemical co-precipitation method and coated with two surfactants to yield a water-based product. The characteristics of the particles were determined via X-ray diffraction (XRD), field emission scanning electron microscopy, and Raman spectrophotometry. The Raman spectra of the BMP showed peaks at 222, 283, 395, 520, 669 and 1316 cm(-1), with broadened band in comparison to the Raman spectra of the magnetic nanoparticles. The BMP absorbance evidenced a rapid increase, with a broad peak at 409 nm, thus reflecting a good loading of the bicalutamide onto the magnetic nanoparticles. The results of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that the MNPs were non-toxic against human brain cancer cells (SH-SY5Y), human cervical cancer cells (Hela), human liver cancer cells (HepG2), breast cancer cells (MCF-7), colon cancer cells (CaCO2) and human prostate cancers (Du 145, PC3) tested herein. In particular, BMPs were cytotoxic at 56% against DU145 cells, at 74.37% in SH-SY5Y cells, and at 58% in Hela cells. Our results demonstrated the biological applicability of BMP nanoparticles as anticancer agents and as agents for enhanced drug delivery against human prostate cancer cells. Our results indicated that the MNPs were biostable and that the BMP functioned effectively as drug delivery vehicles.


Assuntos
Anilidas/síntese química , Antineoplásicos/administração & dosagem , Portadores de Fármacos , Magnetismo , Nanopartículas , Nitrilas/síntese química , Compostos de Tosil/síntese química , Anilidas/química , Antineoplásicos/química , Antineoplásicos/farmacologia , Microscopia Eletrônica de Varredura , Nitrilas/química , Análise Espectral Raman , Compostos de Tosil/química , Difração de Raios X
19.
J Nanosci Nanotechnol ; 10(10): 6962-6, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21137834

RESUMO

Prostate cancer is one of the most preventable cancers, yet effective therapeutics, especially targeted therapy, are still lacking. Saponin has been reported to possess various biological properties such as anti-cancer and anti-inflammatory activity. In this study, the anti-tumor activity of magnetic nanoparticles loaded with ethylene glycol and a saponin complex (ESMP) against DU145 prostate cancer cells was examined. Composite nanoparticles with an average size of 23 nm were prepared using a chemical co-precipitation technique. The ESMP were cytotoxic to DU145 cells and specifically inhibited cell proliferation. In contrast, the magnetic nanoparticles by themselves showed no significant cytotoxicity. The expression levels of the angiogenesis related proteins, including phospho-extracellular signal-regulated kinase, p38 and pAKT, decreased after ESMP treatment. This study highlights the therapeutic potential of using ESMP for targeted-therapy against prostate cancer.


Assuntos
Etilenoglicóis/química , Etilenoglicóis/farmacologia , Nanopartículas de Magnetita/química , Nanocompostos/química , Neoplasias da Próstata/tratamento farmacológico , Saponinas/química , Saponinas/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Etilenoglicóis/síntese química , Humanos , Masculino , Neovascularização Patológica/tratamento farmacológico , Tamanho da Partícula , Neoplasias da Próstata/irrigação sanguínea , Neoplasias da Próstata/patologia , Saponinas/síntese química , Análise Espectral Raman
20.
Nanotechnology ; 18(46): 465201, 2007 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-21730469

RESUMO

This study examined the optical properties of an oxidized form of maghemite (γ-Fe(2)O(3)) nanoparticles and their protective effects against the photoaging of human skin fibroblasts irradiated with ultraviolet (UV) light. Nanoparticles with diameters ranging from 8.7 to 12 nm were prepared using a chemical co-precipitation method. The nanoparticles were coated with two surfactants to obtain a water-based product. The onset of the absorption of the γ-Fe(2)O(3) nanoparticles in the UV-visible absorption spectra increased with increasing particle size. The γ-Fe(2)O(3) nanoparticles significantly inhibited the production of matrix metalloproteinase-1 in human skin fibroblast HS 68 cells by 60% compared with the UV-irradiated control. These results suggest that γ-Fe(2)O(3) nanoparticles have photoprotective properties, and have potential use as an agent against photoaging.

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