RESUMO
The majority of patients with advanced colorectal cancer have chemoresistance to oxaliplatin, and studies on oxaliplatin resistance are limited. Our research showed that RNA-binding motif single-stranded interacting protein 1 (RBMS1) caused ferroptosis resistance in tumor cells, leading to oxaliplatin resistance. We employed bioinformatics to evaluate publically accessible data sets and discovered that RBMS1 was significantly upregulated in oxaliplatin-resistant colorectal cancer cells, in tandem with ferroptosis suppression. In vivo and in vitro studies revealed that inhibiting RBMS1 expression caused ferroptosis in colorectal cancer cells, restoring tumor cell sensitivity to oxaliplatin. Mechanistically, this is due to RBMS1 inducing prion protein translation, resulting in ferroptosis resistance in tumor cells. Validation of clinical specimens revealed that RBMS1 is similarly linked to tumor development and a poor prognosis. Overall, RBMS1 is a potential therapeutic target with clinical translational potential, particularly for oxaliplatin chemoresistance in colorectal cancer.
Assuntos
Neoplasias Colorretais , Ferroptose , Humanos , Oxaliplatina/farmacologia , Resistencia a Medicamentos Antineoplásicos , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Linhagem Celular Tumoral , Proteínas de Ligação a DNA/metabolismo , Proteínas de Ligação a RNA , Proteínas Priônicas/metabolismoRESUMO
Diabetic cardiomyopathy (DCM) is a common complication in patients with diabetes, and ultimately leads to heart failure. Endoplasmic reticulum stress (ERS) induced by abnormal glycolipid metabolism is a critical factor that affects the occurrence and development of DCM. Additionally, the upregulation/activation of silent information regulation 2 homolog-1 (SIRT1) has been shown to protect against DCM. Tanshinone II A (Tan IIA), the main active component of Salviae miltiorrhizae radix et rhizome (a valuable Chinese medicine), has protective effects against cardiovascular disease and diabetes. However, its role and mechanisms in diabetes-induced cardiac dysfunction remain unclear. Therefore, we explored whether Tan IIA alleviates ERS-mediated DCM via SIRT1 and elucidated the underlying mechanism. The results suggested that Tan IIA alleviated the pathological changes in the hearts of diabetic mice, ameliorated the cytopathological morphology of cardiomyocytes, reduced the cell death rate, and inhibited the expression of ERS-related proteins and mRNA. The SIRT1 agonist inhibited the activities of glucose-regulated protein 78 (GRP78). Furthermore, the opposite results under the SIRT1 inhibitor. SIRT1 knockdown was induced by siRNA-SIRT1 transfection, and the degree of GRP78 acetylation was increased. Cumulatively, Tan IIA ameliorated DCM by inhibiting ERS and upregulating SIRT1 expression.
Assuntos
Diabetes Mellitus Experimental , Cardiomiopatias Diabéticas , Humanos , Camundongos , Animais , Miócitos Cardíacos/metabolismo , Cardiomiopatias Diabéticas/tratamento farmacológico , Chaperona BiP do Retículo Endoplasmático , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Sirtuína 1/metabolismo , Estresse do Retículo EndoplasmáticoRESUMO
ABSTRACT: Essential oil from fructus of Alpinia zerumbet (EOFAZ) protects vascular endothelial cell (VEC) injury. Stimulation and injury factors can induce phenotypic changes in VECs and the occurrence of endothelial-mesenchymal transformation (EndMT), accelerating the occurrence and development of cardiovascular diseases. We investigated the role of EOFAZ in EndMT induced by transforming growth factor-ß1 (TGF-ß1). All experiments were performed using human umbilical vein endothelial cells (HUVECs). HUVECs were preincubated with EOFAZ for 2 hours and then coincubated with TGF-ß1 for 72 hours. Krüpple-like factor 4 (KLF4) was inhibited by small interfering RNA or overexpressed by adenovirus infection. Wound healing, transwell, and angiogenesis assays were used to evaluate the migration ability of HUVECs. Quantitative RT-PCR and Western blotting were used for mRNA and protein expression analyses, respectively. Immunofluorescence staining was used to detect expression of related markers. A coimmunoprecipitation assay verified the interaction between KLF4 and acetylated histone H3. TGF-ß1 contributed to EndMT in HUVECs in a time-dependent manner, mainly manifested as an increase in cell migration ability and changes in the expression of EndMT-related mRNAs and proteins. EOFAZ could inhibit EndMT induced by TGF-ß1. The results after transfection with siKLF4 were similar to those of EOFAZ treatment. After EOFAZ treatment, the expression of KLF4 and acetylated histone H3 decreased, and protein interactions between them decreased, while expression of the Notch/Snail signal axis decreased. EOFAZ can attenuate endothelial injuries and suppress EndMT in HUVECs under TGF-ß1 stimulation conditions because it may downregulate KLF4, decrease histone H3 acetylation, and inhibit the transduction of the Notch/Snail signaling axis.
Assuntos
Alpinia , Óleos Voláteis , Fator de Crescimento Transformador beta1 , Alpinia/metabolismo , Regulação para Baixo , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Histonas/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Fator 4 Semelhante a Kruppel/metabolismo , Óleos Voláteis/farmacologia , Fator de Crescimento Transformador beta1/metabolismoRESUMO
Gastrodia elata Bl. has a long edible history and is considered an important functional food raw material. Gastrodin (GAS) is one of the main functional substances in G. elata BI. and can be used as a health care product for the elderly to enhance resistance and delay aging. This study investigated the ameliorative effect and mechanism of GAS on cognitive dysfunction in vascular dementia (VaD) rats, which provides a theoretical basis for development and utilization of functional food. The water maze test shows that GAS improves learning and memory impairment in VaD rats. Meanwhile; GAS significantly decreased the levels of Fe2+ and malondialdehyde (MDA); increased the content of glutathione (GSH); and significantly up-regulated the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and glutathione peroxidase 4 (GPx4), the key regulatory factors of ferroptosis; while it down-regulated the expression of kelch-like ECH-associated protein (Keap1) and cyclooxygenase 2 (COX2). However, GAS does not directly regulate GPx4 and COX2 to inhibit ferroptosis. Furthermore, compared with GAS alone, GAS combined with Bardoxolone (an agonist of Nrf2) did not further affect the increase in GPx4 levels and decrease in COX2 levels, nor did it further affect the regulation of GAS on the biochemical parameters of ferroptosis in HT22 hypoxia injury. These findings revealed that GAS inhibited ferroptosis in hippocampal neurons by activating the Nrf2/Keap1-GPx4 signaling pathway, suggesting its possible application as a functional food for improving vascular dementia by inhibiting ferroptosis.
Assuntos
Disfunção Cognitiva , Demência Vascular , Ferroptose , Animais , Álcoois Benzílicos , Disfunção Cognitiva/tratamento farmacológico , Disfunção Cognitiva/etiologia , Ciclo-Oxigenase 2/metabolismo , Demência Vascular/tratamento farmacológico , Glucosídeos , Glutationa/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Malondialdeído , Fator 2 Relacionado a NF-E2/metabolismo , Ácido Oleanólico/análogos & derivados , Fosfolipídeo Hidroperóxido Glutationa Peroxidase , Ratos , Transdução de SinaisRESUMO
Bacterial infections has become an intractable problem to human health. To overcome this challenge, we developed an antimicrobial agent (AgNPs@PDPE) via the conjugation of a pH-responsive copolymer of PDMAEMA-b-PPEGMA onto AgNPs surface. The AgNPs@PDPE underwent an acidity-induced surface charge conversion that favored bacteria-specific aggregation and antibacterial activity improvement. The specific interaction between AgNPs@PDPE and bacteria under acidic conditions was confirmed via an electrochemical method using AgNPs@PDPE modified glassy carbon electrode as the working electrode. AgNPs@PDPE could efficiently aggregate and inhibit the growth of both Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) under acidic condition. The AgNPs@PDPE could also selectively distinguish pathogenic bacteria from host cells, and this characteristic is benefical for reducing the damage to surrounding tissues in the host. Moreover, AgNPs@PDPE could promote the healing of E. coli- and S. aureus-induced infection, as proven by the histological and TNF-α immunohistochemical analyses of rat dermal wounds. The proposed antimicrobial agent could to be an alternative treatment strategy for the safe treatment of treat bacteria-induced infections in clinics.
Assuntos
Ácidos/farmacologia , Antibacterianos/farmacologia , Escherichia coli/efeitos dos fármacos , Nanopartículas Metálicas/administração & dosagem , Nanocompostos/administração & dosagem , Prata/química , Staphylococcus aureus/efeitos dos fármacos , Ácidos/química , Animais , Escherichia coli/crescimento & desenvolvimento , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Nanopartículas Metálicas/química , Testes de Sensibilidade Microbiana , Nanocompostos/química , Ratos , Ratos Wistar , Staphylococcus aureus/crescimento & desenvolvimentoRESUMO
BACKGROUND Oxymatrine (OMT), a quinolizidine alkaloid derived from the traditional Chinese herb Radix Sophorae flavescentis, has widely reported pharmacological efficacy in treating cardiovascular dysfunction-related diseases. However, the underlying mechanism has been unclear. Here, we investigated the potential inhibitory effects and mechanism of OMT on transdifferentiation of cardiac fibroblast to myofibroblasts induced by aldosterone in vitro. MATERIAL AND METHODS The cardiac fibroblasts (CFBs) proliferation and migration capacity were evaluated by MTT assay, cell cycle assay, and scratch analysis, respectively. The protein expression of the Nrf2/Keap1 signal pathway, FN, Collagen I, Collagen III, alpha-SMA, CTGF, and mineralocorticoid receptor (MR) protein was detected by Western blot analysis. The mRNA expression of Nrf2 was detected by qRT-PCR. Immunofluorescence staining was used to observe the expression of alpha-SMA protein. Nrf2 siRNA was used to explore the role of Nrf2 in OMT-treated CFBs. GSH, SOD, and MDA levels and hydroxyproline content were measured by colorimetric assay with commercial kits. The DCFH-DA fluorescent probe was used to assess cellular ROS levels. RESULTS OMT and Curcumin (an Nrf2 agonist) attenuated aldosterone (ALD)-induced proliferation and migration in CFBs, as well as the fibrosis-associated protein expression levels. Moreover, OMT activated Nrf2 and promoted the nucleus translocation of Nrf2. OMT alleviated the elevated levels of alpha-SMA, Collagen I, Collagen III, and CTGF, which were abrogated by the Nrf2 siRNA transfection. We also found that OMT decreased oxidative stress levels. CONCLUSIONS Our results confirm that OMT alleviates transdifferentiation of cardiac fibroblasts to myofibroblasts induced by aldosterone via activating the Nrf2/Keap1 pathway in vitro.
Assuntos
Alcaloides/farmacologia , Transdiferenciação Celular/efeitos dos fármacos , Miofibroblastos/efeitos dos fármacos , Quinolizinas/farmacologia , Aldosterona/metabolismo , Alcaloides/metabolismo , Animais , Animais Recém-Nascidos , Diferenciação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Colágeno/metabolismo , Fibroblastos/metabolismo , Fibrose , Coração/efeitos dos fármacos , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Miocárdio/patologia , Miofibroblastos/metabolismo , Miofibroblastos/fisiologia , Fator 2 Relacionado a NF-E2/metabolismo , Quinolizinas/metabolismo , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosRESUMO
Antimicrobial resistance to traditional antibiotics leads to a serious concern for medical care owing to ineffective antibiotic therapies. This study focused on the preparation of silver nanocomposites (AgNPs@Tob&PAGA) by modifying AgNPs with tobramycin (Tob) and carbohydrate polymer of poly(2-(acrylamido) glucopyranose) (PAGA). The enhanced antibacterial activities of nanocomposites against common pathogens of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were explored. The introduction of PAGA onto silver nanocomposites improved both citocompatibility and antibacterial activity. Compared with nude Tob, AgNPs@Tob&PAGA showed more fascinating antimicrobial effect against E. coli and S. aureus with about 20-fold increase in the antibacterial activity, simultaneously no detectable resistance was observed. Consequently, the silver nanocomposite as an antimicrobial agent presents promising prospects in the treatment of bacterial infections caused by antimicrobial resistant bacteria.
Assuntos
Antibacterianos/química , Farmacorresistência Bacteriana , Nanopartículas Metálicas/química , Nanocompostos/química , Ácido Poliglicólico/análogos & derivados , Prata/química , Animais , Materiais Biocompatíveis/química , Carboidratos/química , Sobrevivência Celular , Escherichia coli/efeitos dos fármacos , Grafite , Células Endoteliais da Veia Umbilical Humana , Humanos , Camundongos , Testes de Sensibilidade Microbiana , Células NIH 3T3 , Ácido Poliglicólico/química , Staphylococcus aureus/efeitos dos fármacos , Tobramicina/químicaRESUMO
BACKGROUND Salvianolic acid B (SalB) is the representative component of phenolic acids derived from the roots and rhizomes of Salvia miltiorrhiza Bge (Labiatae), which has been used widely in Asian countries for clinical therapy of various cardiovascular dysfunction-related diseases. However, cardiac protection effects and the underlying mechanism for clinical application are still poorly understood. Here, we investigated the potential anti-myocardial fibrosis effect and mechanism of SalB on Angiotensin II (Ang II)-induced cardiac fibrosis in vitro. MATERIAL AND METHODS The proliferation and migration capacity of cardiac fibroblasts (CFBs) were measured by MTT assay and scratch analysis, respectively. The colorimetric assay determined the hydroxyproline content in medium. Western blotting detected the protein expressions of nuclear transcription factor-kappa B (NF-κB) pathway-associated proteins, fibronectin (FN), collagen type I (Coll I), α-smooth muscle actin (α-SMA), and connective tissue growth factor (CTGF). The expression of α-SMA protein was observed by immunofluorescence staining. qRT-PCR detected the mRNA expression of NF-κB. RESULTS SalB attenuated Ang II-induced the proliferation and the migration ability of CFBs. Ang II-induced the extracellular matrix protein Coll I, FN, and α-SMA, the pro-fibrotic cytokine CTGF protein expression was inhibited, and the nuclear translocation of NF-κB p65 subunit was reduced by SalB. Western blotting and qRT-PCR confirmed that SalB blocked the activation of NF-κB induced by Ang II. PDTC (the NF-κB inhibitor) also inhibited proliferation of CFBs and reduced α-SMA and Coll I expression induced by Ang II. CONCLUSIONS SalB can alleviate Ang II-induced cardiac fibrosis via suppressing the NF-κB pathway in vitro.
Assuntos
Benzofuranos/farmacologia , Fibroblastos/efeitos dos fármacos , Miocárdio/metabolismo , Angiotensina II/metabolismo , Animais , Benzofuranos/metabolismo , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Colágeno Tipo I/metabolismo , Fibrose/tratamento farmacológico , Coração/efeitos dos fármacos , Proteínas I-kappa B/metabolismo , Miocárdio/patologia , NF-kappa B/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND Oxymatrine, a component extracted from the traditional Chinese herb Sophora japonica (Sophora flavescens Ait.), has various pharmacological effects, especially on the cardiovascular system. However, its cardiac protection effects and the underlying mechanism are still poorly understood. In the present study, we investigated the inhibitory effect and mechanism of oxymatrine on cardiac fibrosis induced by TGF-ß1. MATERIAL AND METHODS Cardiac fibroblasts were isolated and purified from neonatal rats. Immunocytochemical staining was used to identify the cells. MTT assay and immunofluorescence staining were used to assess cardiac fibroblasts proliferation and myofibroblasts transformation. Hematoxylin-eosin staining was performed to evaluate morphological changes of cardiac fibroblasts. The secretion of type I and III collagen was assessed by staining with picrosirius red and the hydroxyproline content was determined by colorimetric assay. Cardiac fibroblast migration was examined by scratch assay and DNA content was detected to analyze cell cycle distribution using flow cytometry. Western blot analysis was used to detect the protein expressions of Notch pathway-associated protein in cardiac fibroblasts. RESULTS Oxymatrine and Notch signaling pathway inhibitor DAPT could attenuated TGF-ß1 induced the capacity of proliferation and migration increased in cardiac fibroblasts, as well as the secretion of collagen and hydroxyproline colorimetric content in medium. TGF-ß1 induced the biomarker α-SMA of fibroblast-to-myofibroblast transformation (FMT), which was inhibited by oxymatrine and DAPT. Western blotting confirmed that oxymatrine blocked the activation of Notch induced by TGF-ß1. CONCLUSIONS Oxymatrine is a potential inhibitor FMT induced by TGF-ß1, which is at least in part mediated via inhibition of Notch signaling.
Assuntos
Alcaloides/farmacologia , Coração/efeitos dos fármacos , Miofibroblastos/efeitos dos fármacos , Quinolizinas/farmacologia , Receptores Notch/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Dipeptídeos/farmacologia , Miocárdio/citologia , Miocárdio/metabolismo , Miofibroblastos/citologia , Miofibroblastos/metabolismo , Fosforilação/efeitos dos fármacos , Cultura Primária de Células , Ratos , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND In China, the essential oil of the fruit, Fructus Alpiniae zerumbet (FAZ), is used to treat cardiovascular diseases. Recent in vitro studies have shown that the essential oil of FAZ (EOFAZ) can protect endothelial cells from injury. Because of the prevalence of diabetes mellitus and its effects on the cardiovascular system, the aim of this study was to investigate the mechanism of the effects of EOFAZ on human umbilical vein endothelial cells (HUVECs) treated with high levels of glucose in vitro. MATERIAL AND METHODS The lactate dehydrogenase (LDH) leakage assay was used to detect HUVEC injury. Tumor necrosis factor-alpha (TNF-α), interleukin-8 (IL-8), and nuclear transcription factor-kappa B (NF-κB) p65 subunit DNA-binding activity was detected. The expression of NF-κB pathway-associated proteins, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) was studied by Western blotting. The cellular location of NF-κB in HUVECs was evaluated using immunofluorescence. RESULTS Cell viability and LDH leakage assays showed that high glucose-induced HUVEC injury was reduced by EOFAZ. High glucose-induced secretion of IL-8, TNF-α, ICAM-1, and VCAM-1 was reduced, and translocation of the p65 subunit of NF-κB to the endothelial cell nucleus was inhibited by EOFAZ. Western blotting confirmed that EOFAZ blocked the activation of NF-κB induced by high glucose levels. EOFAZ reduced high glucose-induced p65/DNA binding to inhibit NF-κB activation. CONCLUSIONS The findings of this in vitro study showed that treatment of HUVECs with EOFAZ had a protective role against the effects of high glucose levels via the NF-κB signaling pathway.
Assuntos
Alpinia/metabolismo , Glucose/efeitos adversos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Cultivadas , China , Endotélio Vascular/efeitos dos fármacos , Frutas , Regulação da Expressão Gênica/efeitos dos fármacos , Glucose/metabolismo , Humanos , Proteínas I-kappa B/metabolismo , Molécula 1 de Adesão Intercelular/biossíntese , Medicina Tradicional Chinesa , NF-kappa B/metabolismo , Óleos Voláteis/farmacologia , Substâncias Protetoras/farmacologia , Transdução de Sinais/efeitos dos fármacos , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Molécula 1 de Adesão de Célula Vascular/biossínteseRESUMO
BACKGROUND: We previously demonstrated oxymatrine, an alkaloid from the Chinese medicine radix Sophorae flavescentis, ameliorates hemodynamic disturbances and cardiac fibrosis; however, the underlying mechanisms are unclear. Here, we investigated the effect and mechanism of action of oxymatrine on aldosterone-induced cardiac fibroblast to myofibroblast differentiation in vitro. METHODS: Cardiac fibroblasts were isolated purified from neonatal Sprague Dawley rats. The optimal concentration of aldosterone to stimulate cardiac fibroblast proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cardiac fibroblasts were pretreated with 7.57 × 10(-4) mol/L or 3.78 × 10(-4) mol/L oxymatrine or without oxymatrine for 2 h, and then coincubated with 1 × 10(-8) mol/L aldosterone for 48 h. The MTT assay and Masson staining were used to detect the cardiac fibroblast proliferation and myofibroblast differentiation. The secretion of type I and III collagen was measured by commercial ELISA kits, and the hydroxyproline content was determined by the colorimetric assay. Western blotting assayed the Smad-2, Smad-3, and Smad-4 protein expression in cardiac fibroblasts. RESULTS: The present results confirmed that aldosterone induced cardiac fibroblast to myofibroblast proliferation and differentiation. The MTT assay and Masson staining indicated oxymatrine significantly inhibited aldosterone-induced cardiac fibroblast proliferation and myofibroblast differentiation. Oxymatrine significantly inhibited aldosterone-induced secretion of type I and III collagen, as indicated by commercial ELISA kits, and aldosterone-induced increase in hydroxyproline content, as indicated by a colorimetric assay. Western blotting revealed oxymatrine attenuated aldosterone-induced Smad-2, Smad-3, and Smad-4 expression in cardiac fibroblasts. CONCLUSION: Oxymatrine can inhibit cardiac fibroblast proliferation and differentiation into myofibroblasts via a mechanism linked to attenuation of the Smad signaling pathway.
Assuntos
Aldosterona/farmacologia , Alcaloides/farmacologia , Proliferação de Células/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Quinolizinas/farmacologia , Proteínas Smad/metabolismo , Alcaloides/química , Animais , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Miocárdio/citologia , Substâncias Protetoras/química , Quinolizinas/química , Ratos , Ratos Sprague-Dawley , Proteínas Smad/análise , Proteínas Smad/genéticaRESUMO
BACKGROUND: To investigate the ameliorated effects of an extract of Ginkgo biloba extract (GBE) on experimental cardiac remodeling in rats induced by acute cardiac infarction, and further explore the mechanism concentrated on myocardial type I collagen, transforming growth factor beta 1 (TGF-ß1), matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9), and provide the experimental data for clinical application of GBE. METHODS: Rats were divided into five groups (n = 20) as following: sham operation group (group A), acute myocardial infarction model group (group B), acute myocardial infarction model + aspirin (10 mg/kg) treatment group (group C), acute myocardial infarction model + captopril (20 mg/kg) treatment group (group D) and acute myocardial infarction model + Ginkgo biloba extract (100 mg/kg) treatment group (group E). The rat acute myocardial infarction model was reproduced by ligaturing the left anterior descending artery excluding the sham operation group which did not ligation only completed the operational process. Each group was further subdivided into treatment regimens lasting 4 weeks and 8 weeks. Immunohistochemistry and real-time polymerase chain reaction (PCR) methods were used to detect the protein expression and mRNA transcriptional levels of rat myocardial TGF-ß1, type I collagen, MMP-2 and MMP-9. RESULTS: Compared with group B, regardless of the length of treatment (4 or 8 weeks), the TGF-ß1, MMP-2 and MMP-9 mRNA transcriptional levels, and the protein expression levels of type I collagen, MMP-2 and MMP-9 in groups D, C and E were significantly decreased (P < 0.01). Furthermore, the mRNA expression levels of TGF-ß1 in groups D, C and E were significantly lower after 8 weeks compared to after 4 weeks (P < 0.01), as were the expression levels of type I collagen in groups D, C and E (P < 0.05). There was no statistically significant difference in the protein expression levels of MMP-2 and MMP-9 between groups E and C. CONCLUSIONS: GBE could inhibit experimental rat myocardial remodeling after acute myocardial infarction via reduced transcription of TGF-ß1, MMP-2 and MMP-9 genes and by the decreased expression of type I collagen, MMP-2 and MMP-9 proteins in myocardial cells.
Assuntos
Ginkgo biloba , Infarto do Miocárdio/metabolismo , Extratos Vegetais/farmacologia , Remodelação Ventricular/efeitos dos fármacos , Animais , Colágeno Tipo I/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Ratos , Fator de Crescimento Transformador beta1/metabolismoRESUMO
OBJECTIVE: To investigate the inhibitory effects of OMT on TGF-ß1-induced CFBs proliferation, and then explore the mechanism. METHOD: The experiment was randomly divided into 6 groups as following: control group (serum free DMEM), model group (20 µg x L(-1) TGF-ß1), OMT low dose group (1.89 x 10(-4) mol x L(-1) + 20 µg x L(-1) TGF-ß1), OMT medium dose group (3.78 x 10(-4) mol x L(-1) + 20 µg x L(-1) TGF-ß1), OMT high dose group (7.56 x 10(-4) mol x L(-1) + 20 µg x L(-1) TGF-ß1), SB203580 group (p38MAPK blocking agent, 1 x 10(-5) mol x L(-1) + 20 µg x L(-1) TGF-ß1). Vimentin of CFBs was identified by immunocytochemical methods, α-SMA of myFBs as well. Inhibitory effects of OMT on CFBs proliferation was detected by the MTT assay. Picric acid Sirius red staining was analyzed collagen type I and collagen type III deposition. Western blot was determined the expression of p38MAPK, p-p38MAPK, collagen type I and collagen type III. RESULT: MTT results showed that OMT significantly inhibited CFBs proliferation induced by TGF-ß1 (P < 0.01) α-SMA immunocytochemical experiments suggested that OMT could protect against the CFBs proliferation. OMT could significantly decrease the deposition of collagen type I and collagen type III by Western bloting and picric acid Sirius red staining. Western blot results showed that TGF-ß1 enhanced p38MAPK phosphorylation, however OMT attenuated the phosphorylation of p38MAPK induced by TGF-ß1 (P < 0.01). CONCLUSION: OMT can inhibit the CFBs proliferation induced by TGF-ß1, and its mechanism may be involved in inhibiting p38MAPK phosphorylation.
Assuntos
Alcaloides/farmacologia , Coração/efeitos dos fármacos , Quinolizinas/farmacologia , Fator de Crescimento Transformador beta1/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Animais , Proliferação de Células/efeitos dos fármacos , Colágeno/metabolismo , Regulação para Baixo , Feminino , Fibroblastos/efeitos dos fármacos , Técnicas In Vitro , Masculino , Fosforilação , Ratos , Ratos Sprague-Dawley , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Oxidized low-density lipoprotein has been demonstrated to induce vascular injury, at least partly, through the nitric oxide-dependent pathway. Our previous studies showed that the essential oil from Fructus Alpiniae zerumbet had a capacity of antioxidation to ameliorate endothelial function and vasodilatation on aortic rings ex vivo. The present study aims to confirm the hypothesis that Fructus Alpiniae zerumbet protects against oxidized low-density lipoprotein-induced endothelial dysfunction in human aortic endothelial cells, and the role nitric oxide synthase plays in the process. Human aortic endothelial cells were pretreated with one of three different doses of Fructus Alpiniae zerumbet (1, 2, or 4 µg/mL), simvastatin (10 µM), or vehicle,followed by coincubation with oxidized low-density lipoprotein (150 µg/mL), for designed time periods. Exposure to oxidized low-density lipoprotein resulted in cytotoxicity accompanied by mitochondrial membrane potential collapse, excessive reactive oxygen species generation, and a decrease of nitric oxide production. The upregulation of inducible nitric oxide synthase mRNA and protein expression was determined by reverse transcription-polymerase chain reaction and Western blot analysis, respectively. Furthermore, oxidized low-density lipoprotein inhibited Akt phosphorylation and downregulated endothelial nitric oxide synthase and soluble guanylatecyclase in mRNA expression and the protein level. However, cytotoxicity and aberrant reactive oxygen species and nitric oxide generation caused by oxidized low-density lipoprotein could be significantly inhibited by Fructus Alpiniae zerumbet or Sim pretreatment. Results from this study confirmed that Fructus Alpiniae zerumbet could protect against oxidized low-density lipoprotein-induced endothelial dysfunction by antioxidation and modulating the Akt/nitric oxide synthase-nitric oxide signaling pathway.
Assuntos
Alpinia/química , Endotélio Vascular/efeitos dos fármacos , Óleos Voláteis/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Aorta/efeitos dos fármacos , Células Cultivadas , Humanos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/metabolismo , Óleos Voláteis/química , Óleos Voláteis/isolamento & purificaçãoRESUMO
OBJECTIVE: To investigate the ameliorated effect of CVB-D on oxidative stress and energy metabolism in experimental cardiac injuried rats induced by sympathetic overactivity in vivo. METHODS: SD rats were randomly divided into five groups as following: control group, model group, Vitamin E 150 mg/kg group, CVB-D low dose and high dose groups, respectively. The rat experimental cardiac injury model was established by exposed to norepinephrine (NE) 3 mg/kg by ip for 16 d. The drugs were administrated to rat for 16 d by ig. The body weight of rats were monitored during all of the experimental period. At the designing ending-time point the indexes were assayed as following: cardiac index, hydroxyproline, histopathologically examination, oxidative stress ( MDA, SOD, CAT, GSH-Px and T-AOC) and energy metabolism indicatricle ( Na+, K(+) -ATPase, and Ca2+, Mg(2+) -ATPase). RESULTS: After exposed with NE for 16 d, the rats of model group was appeared dysfunction of oxidative stress and energy metabolism such as decreasing body weight, increasing cardiac index and hydroxyproline in cardiac tissue, decreasing Na+, K(+) -ATPase and Ca(2+), Mg(2+) -ATPase activities, and deteriorating the oxidative stress. Treated with CVB-D could ameliorate all of the exacerbated indexes. CONCLUSION: CVB-D has protective effect against oxidative stress and energy metabolism in rats of experimental myocardial injury induced by sympathetic overactivity.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Metabolismo Energético/efeitos dos fármacos , Coração/efeitos dos fármacos , Coração/fisiopatologia , Estresse Oxidativo/efeitos dos fármacos , Adenosina Trifosfatases , Animais , Traumatismos Cardíacos , Ratos , Ratos Sprague-Dawley , Vitamina ERESUMO
OBJECTIVE: To investigate the protective effect of CVB-D on cardiac myocytes injury induced by high sympathesis activity and its relationship with oxidative stress. METHODS: Primary culture cardiac myocyte of new-born rat was injuried by NE and then incubated with VE and CVB-D (10 and 50 micromol/L). Indexes of cardiac myoctye injury were assayed by morphologic change, MTT, and LDH leakage ratio. The activity of SOD and the content of MDA were investigated to identify oxidation and antioxygen. RESULTS: CVB-D (10 and 50 micromol/L) significantly increased the cell survival rate,and reduced the LDH leakage rate. CVE-D (50 micromol/L) significantly increased the activity of SOD, and decreased content of MDA in injuried cell. CONCLUSION: CVB-D has protective effect against myocardial injury induced by high sympathesis activity, the mechanism involves in ameliorating oxidative stress.
Assuntos
Miócitos Cardíacos , Animais , Sobrevivência Celular , Células Cultivadas , Traumatismos Cardíacos , Estresse Oxidativo , RatosRESUMO
OBJECTIVE: Enterotypes (ETs) are the clustering of gut microbial community structures, which could serve as indicators of growth performance and carcass traits. However, ETs have been sparsely investigated in waterfowl. The objective of this study was to identify the ileal ETs and explore the correlation of the ETs with growth performance and carcass traits in Muscovy ducks. METHODS: A total of 200 Muscovy ducks were randomly selected from a population of 5,000 ducks at 70-day old, weighed and slaughtered. The growth performance and carcass traits, including body weight, dressed weight and evidenced weight, dressed percentage, percentage of apparent yield, breast muscle weight, leg muscle weight, percentage of leg muscle and percentage of breast muscle, were determined. The contents of ileum were collected for the isolation of DNA and 16S rRNA gene sequencing. The ETs were identified based on the 16S rRNA gene sequencing data and the correlation of the ETs with growth performance and carcass traits was performed by Spearman correlation analysis. RESULTS: Three ETs (ET1, ET2, and ET3) were observed in the ileal microbiota of Muscovy ducks with significant differences in number of features and α-diversity among these ETs (p<0.05). Streptococcus, Candida Arthritis, and Bacteroidetes were the presentative genus in ET1 to ET3, respectively. Correlation analysis revealed that Lactococcus and Bradyrhizobium were significantly correlated with percentage of eviscerated yield and leg muscle weight (p<0.05) while ETs were found to have a close association with percentage of eviscerated yield, leg muscle weight, and percentage of leg muscle in Muscovy ducks. However, the growth performance of ducks with different ETs did not show significant difference (p>0.05). Lactococcus were found to be significantly correlated with leg muscle weight, dressed weight, and percentage of eviscerated yield. CONCLUSION: Our findings revealed a substantial variation in carcass traits associated with ETs in Muscovy ducks. It is implied that ETs might have the potential to serve as a valuable biomarker for assessing duck carcass traits. It would provide novel insights into the interaction of gut microbiota with growth performance and carcass traits of ducks.
RESUMO
Accumulating evidence strongly support the key role of NLRP3-mediated pyroptosis in the pathogenesis and progression of vascular endothelial dysfunction associated with diabetes mellitus. Various studies have demonstrated that the activation or upregulation of Silent Information Regulation 2 homolog 2 (SIRT2) exerts inhibitory effect on the expression of NLRP3. Although 1,8-cineole has been found to protect against endothelial dysfunction and cardiovascular diseases, its role and mechanism in diabetic angiopathy remain unknown. Therefore, the aim of this study was to investigate the ameliorative effect of 1,8-cineole through SIRT2 on pyroptosis associated with diabetic angiopathy in human umbilical vein endothelial cells (HUVECs) and to elucidate the underlying mechanism. The findings revealed that 1,8-cineole exhibited a protective effect against vascular injury and ameliorated pathological alterations in the thoracic aorta of diabetic mice. Moreover, it effectively mitigated pyroptosis induced by palmitic acid-high glucose (PA-HG) in HUVECs. Treatment with 1,8-cineole effectively restored the reduced levels of SIRT2 and suppressed the elevated expression of pyroptosis-associated proteins. Additionally, our findings demonstrated the occurrence of NLRP3 deacetylation and the physical interaction between NLRP3 and SIRT2. The SIRT2 inhibitor AGK2 and siRNA-SIRT2 effectively attenuated the effect of 1,8-cineole on NLRP3 deacetylation in HUVECs and compromised its inhibitory effect against pyroptosis in HUVECs. However, overexpression of SIRT2 inhibited PA-HG-induced pyroptosis in HUVECs. 1,8-Cineole inhibited the deacetylation of NLRP3 by regulating SIRT2, thereby reducing pyroptosis in HUVECs. In conclusion, our findings suggest that PA-HG-induced pyroptosis in HUVECs plays a crucial role in the development of diabetic angiopathy, which can be mitigated by 1,8-cineole.
Assuntos
Diabetes Mellitus Experimental , Eucaliptol , Células Endoteliais da Veia Umbilical Humana , Inflamassomos , Piroptose , Animais , Humanos , Masculino , Camundongos , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Angiopatias Diabéticas/tratamento farmacológico , Angiopatias Diabéticas/metabolismo , Angiopatias Diabéticas/prevenção & controle , Angiopatias Diabéticas/patologia , Eucaliptol/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Inflamassomos/metabolismo , Inflamassomos/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Piroptose/efeitos dos fármacos , Sirtuína 2/metabolismo , Sirtuína 2/antagonistas & inibidoresRESUMO
BACKGROUND: Atherosclerosis (AS) is a progressive chronic disease. Currently, cardiovascular diseases (CVDs) caused by AS is responsible for the global increased mortality. Yanshanjiang as miao herb in Guizhou of China is the dried and ripe fruit of Fructus Alpinia zerumbet. Accumulated evidences have confirmed that Yanshanjiang could ameliorate CVDs, including AS. Nevertheless, its effect and mechanism on AS are still largely unknown. PURPOSE: To investigate the role of essential oil from Fructus Alpinia zerumbet (EOFAZ) on AS, and the potential mechanism. METHODS: A high-fat diet (HFD) ApoE-/- mice model of AS and a oxLDL-induced model of macrophage-derived foam cells (MFCs) were reproduced to investigate the pharmacological properties of EOFAZ on AS in vivo and foam cell formation in vitro, respectively. The underlying mechanisms of EOFAZ were investigated using Network pharmacology and molecular docking. EOFAZ effect on PPARγ protein stability was measured using a cellular thermal shift assay (CETSA). Pharmacological agonists and inhibitors and gene interventions were employed for clarifying EOFAZ's potential mechanism. RESULTS: EOFAZ attenuated AS progression in HFD ApoE-/- mice. This attenuation was manifested by the reduced aortic intima plaque development, increased collagen content in aortic plaques, notable improvement in lipid profiles, and decreased levels of inflammatory factors. Moreover, EOFAZ inhibited the formation of MFCs by enhancing cholesterol efflux through activiting the PPARγ-LXRα-ABCA1/G1 pathway. Interestingly, the pharmacological knockdown of PPARγ impaired the beneficial effects of EOFAZ on MFCs. Additionally, our results indicated that EOFAZ reduced the ubiquitination degradation of PPARγ, and the chemical composition of EOFAZ directly bound to the PPARγ protein, thereby increasing its stability. Finally, PPARγ knockdown mitigated the protective effects of EOFAZ on AS in HFD ApoE-/- mice. CONCLUSION: These findings represent the first confirmation of EOFAZ's in vivo anti-atherosclerotic effects in ApoE-/- mice. Mechanistically, its chemical constituents can directly bind to PPARγ protein, enhancing its stability, while reducing PPARγ ubiquitination degradation, thereby inhibiting foam cell formation via activation of the PPARγ-LXRα-ABCA1/G1 pathway. Simultaneously, EOFAZ could ameliorates blood lipid metabolism and inflammatory microenvironment, thus synergistically exerting its anti-atherosclerotic effects.
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Alpinia , Aterosclerose , Óleos Voláteis , Placa Aterosclerótica , Animais , Camundongos , PPAR gama/metabolismo , Óleos Voláteis/farmacologia , Frutas , Simulação de Acoplamento Molecular , Transdução de Sinais , Aterosclerose/tratamento farmacológico , Aterosclerose/metabolismo , Placa Aterosclerótica/tratamento farmacológico , Apolipoproteínas E , Transportador 1 de Cassete de Ligação de ATP/metabolismo , Receptores X do Fígado/metabolismoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Vascular endothelial cell senescence is associated with cardiovascular complications in diabetes. Essential oil from Fructus Alpiniae zerumbet (Pers.) B.L.Burtt & R.M.Sm. (EOFAZ) has potentially beneficial and promising diabetes-related vascular endothelial cell senescence-mitigating effects; however, the underlying molecular mechanisms remain unclear. AIM OF THE STUDY: To investigate the molecular effects of EOFAZ on vascular endothelial cell senescence in diabetes. MATERIALS AND METHODS: A diabetes mouse model was developed using a high-fat and high-glucose diet (HFD) combined with intraperitoneal injection of low-dose streptozotocin (STZ, 30 mg/kg) and oral treatment with EOFAZ. 4D label-free quantitative proteomics, network pharmacology, and molecular docking techniques were employed to explore the molecular mechanisms via which EOFAZ alleviates diabetes-related vascular endothelial cell senescence. A human aortic endothelial cells (HAECs) senescence model was developed using high palmitic acid and high glucose (PA/HG) concentrations in vitro. Western blotting, immunofluorescence, SA-ß-galactosidase staining, cell cycle, reactive oxygen species (ROS), cell migration, and enzyme linked immunosorbent assays were performed to determine the protective role of EOFAZ against vascular endothelial cell senescence in diabetes. Moreover, the PPAR-γ agonist rosiglitazone, inhibitor GW9662, and siRNA were used to verify the underlying mechanism by which EOFAZ combats vascular endothelial cell senescence in diabetes. RESULTS: EOFAZ treatment ameliorated abnormal lipid metabolism, vascular histopathological damage, and vascular endothelial aging in diabetic mice. Proteomics and network pharmacology analysis revealed that the differentially expressed proteins (DEPs) and drug-disease targets were associated with the peroxisome proliferator-activated receptor gamma (PPAR-γ) signalling pathway, a key player in vascular endothelial cell senescence. Molecular docking indicated that the small-molecule compounds in EOFAZ had a high affinity for the PPAR-γ protein. Western blotting and immunofluorescence analyses confirmed the significance of DEPs and the involvement of the PPAR-γ signalling pathway. In vitro, EOFAZ and rosiglitazone treatment reversed the effects of PA/HG on the number of senescent endothelial cells, expression of senescence-related proteins, the proportion of cells in the G0/G1 phase, ROS levels, cell migration rate, and expression of pro-inflammatory factors. The protective effects of EOFAZ against vascular endothelial cell senescence in diabetes were aborted following treatment with GW9662 or PPAR-γ siRNA. CONCLUSIONS: EOFAZ ameliorates vascular endothelial cell senescence in diabetes by activating PPAR-γ signalling. The results of the present study highlight the potential beneficial and promising therapeutic effects of EOFAZ and provide a basis for its clinical application in diabetes-related vascular endothelial cell senescence.