RESUMO
OBJECTIVE AND DESIGN: Ulcerative colitis (UC) is a multi-faceted, recurrent immune disorder caused by dextran sulfate sodium (DSS). The intestinal microbiota has multiple functions in the host, so UC requires long-term potent medication. The effect of resveratrol (RSV) has seldom been reported, and this study researched that. Herein, the effect of RSV and Grape seed oil that anti-inflammatory ability in experimental mice was explored, also why RSV altered Gut Microbiota has been researched. MATERIALS AND METHODS: In this experiment, the effects of experimental drugs on colon length in mice with DSS-induced colitis were compared. H&E Staining was performed on serial sections of colon tissues and histological scores were determined for all groups. The expression of cyclooxygenase-2 (COX-2) and tumor necrosis factor-α (TNF-α) in the colon tissue of mice was detected by immunohistochemical staining. In the end, the α-diversity index, sobs index, and rarefaction curve of the cecal and colon microbiota of different groups of mice were measured. Bray-Curtis-based Venn diagram of PCoA (principal coordinate analysis) and OTUs distribution in mouse gut microbiota were obtained. RESULTS: The results showed that the use of 40 mg/kg RSV (high dose) significantly reduced the severity of UC. The use of 10 mg/kg RSV (low dose) significantly reduced the effect of shortened colon length in DSS mice. Compared with the DSS-treated group, the levels of COX-2 and TNF-α in the colon tissues of RSV + DSS-treated mice were significantly decreased. According to this experiment, 19 mouse gut microbiota species had a relative abundance greater than 0.1%, with Beerella, Bacteroides, Helicobacter, Oscillator, and cecum pylori being more abundant in the colon than in the colon. A higher relative abundance of Lachnospira NK4A136 was observed in DSS and RSV groups compared with the control group, whereas the opposite was observed for Alloprevotella. This proves that resveratrol increases the uniformity and diversity of gut microbes to a certain extent, and has a protective effect on the gut.
Assuntos
Colite Ulcerativa , Sulfato de Dextrana , Microbioma Gastrointestinal , Resveratrol , Animais , Resveratrol/farmacologia , Microbioma Gastrointestinal/efeitos dos fármacos , Colite Ulcerativa/tratamento farmacológico , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/microbiologia , Colite Ulcerativa/patologia , Camundongos , Masculino , Ciclo-Oxigenase 2/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Colo/efeitos dos fármacos , Colo/patologia , Colo/metabolismo , Camundongos Endogâmicos C57BL , Modelos Animais de Doenças , Anti-Inflamatórios/farmacologiaRESUMO
OBJECTIVES: Chronic neuroinflammation has become one of the important causes of common neurodegeneration disease. Therefore, the target of this study was to explore the protective action of glabridin on lipopolysaccharide (LPS)-induced neuroinflammation in vivo and in vitro and its mechanism. METHODS: The neuroinflammation model was established by LPS-induced BV2 cells. The cell viability with various concentrations of glabridin was determined by MTT assay, and the content of NO in each group was detected. A neuroinflammatory model was established in male C57BL/6J mice for a water maze test. Subsequently, NF-κB and SOD indices were measured by ELISA, GFAP and IBA-1 indices were measured by immunofluorescence, and Nissl staining was used to explore the Nissl bodies in the hippocampus of mice. RESULTS: In vitro experiments, our results expressed that glabridin could markedly increase the cell activity of LPS-induced BV2 cells and reduce the NO expression in cells. It indicated that glabridin had a remarkable impact on the neuroinflammation of LPS-induced BV2 cell protection. In vivo neuroinflammation experiments, mice treated with different doses of glabridin showed significantly improved ability of memory compared with the LPS group in the Morris water maze test. The levels of NF-κB, GFAP, and the number of positive cells in Nissl staining were decreased. High-dose glabridin significantly increased the SOD content in the brain tissue and decreased the IBA-1 levels. CONCLUSION: Glabridin can significantly reduce or even reverse LPS-induced neuroinflammation, which may be related to the fact that glabridin can reduce the NO expression, NF-κB, IBA-1, GFAP, and other inflammatory mediators, upregulate the expression of SOD to relieve oxidative stress of brain and inhibit the activation of gliocyte in brain tissue.
Assuntos
Isoflavonas , NF-kappa B , Fenóis , Transdução de Sinais , Camundongos , Animais , Masculino , NF-kappa B/metabolismo , Lipopolissacarídeos/farmacologia , Doenças Neuroinflamatórias , Inflamação/metabolismo , Camundongos Endogâmicos C57BL , Superóxido Dismutase/metabolismo , Microglia/metabolismoRESUMO
The principal aim of present study was to assess the therapeutic efficacy of bone morphogenetic protein-7 (BMP-7) induced differentiation of bone marrow mesenchymal stem cells (BMSCs) in a rat acute spinal cord injury (SCI) model. BMSCs were isolated from rats, and then divided into a control and a BMP-7 induction groups. The proliferation ability of BMSCs and glial cell markers were determined. Forty Sprague-Dawley (SD) rats were randomly divided into sham, SCI, BMSC, and BMP7 + BMSC groups (n = 10). Among these rats, the recovery of hind limb motor function, the pathological related markers, and motor evoked potentials (MEP) were identified. BMSCs differentiated into neuron-like cells after the introduction of exogenous BMP-7. Interestingly, the expression levels of MAP-2 and Nestin increased, whereas the expression level of GFAP decreased after the treatment with exogenous BMP-7. Furthermore, the Basso, Beattie, and Bresnahan (BBB) score reached 19.33 ± 0.58 in the BMP-7 + BMSC group at day 42. Nissl bodies in the model group were reduced compared to the sham group. After 42 days, in both the BMSC and BMP-7 + BMSC groups, the number of Nissl bodies increased. This is especially so for the number of Nissl bodies in the BMP-7 + BMSC group, which was more than that in the BMSC group. The expression of Tuj-1 and MBP in BMP-7 + BMSC group increased, whereas the expression of GFAP decreased. Moreover, the MEP waveform decreased significantly after surgery. Furthermore, the waveform was wider and the amplitude was higher in BMP-7 + BMSC group than that in BMSC group. BMP-7 promotes BMSC proliferation, induces the differentiation of BMSCsinto neuron-like cells, and inhibits the formation of glial scar. BMP-7 plays a confident role in the recovery of SCI rats.
Assuntos
Células-Tronco Mesenquimais , Traumatismos da Medula Espinal , Ratos , Animais , Proteína Morfogenética Óssea 7/genética , Ratos Sprague-Dawley , Diferenciação Celular , Traumatismos da Medula Espinal/terapiaRESUMO
BACKGROUND: Cervical cancer remains one of the most prevalent cancers worldwide. Accumulating evidence suggests that specificity protein 1 (Sp1) plays a pivotal role in tumour progression. The underlying role and mechanism of Sp1 in tumour progression remain unclear. METHODS: The protein level of Sp1 in tumour tissues was determined by immunohistochemistry. The effect of Sp1 expression on the biological characteristics of cervical cancer cells was assessed by colony, wound healing, transwell formation, EdU, and TUNEL assays. Finally, the underlying mechanisms and effects of Sp1 on the mitochondrial network and metabolism of cervical cancer were analysed both in vitro and in vivo. RESULTS: Sp1 expression was upregulated in cervical cancer. Sp1 knockdown suppressed cell proliferation both in vitro and in vivo, while overexpression of Sp1 had the opposite effects. Mechanistically, Sp1 facilitated mitochondrial remodelling by regulating mitofusin 1/2 (Mfn1/2), OPA1 mitochondrial dynamin-like GTPase (Opa1), and dynamin 1-like (Drp1). Additionally, the Sp1-mediated reprogramming of glucose metabolism played a critical role in the progression of cervical cancer cells. CONCLUSIONS: Our study demonstrates that Sp1 plays a vital role in cervical tumorigenesis by regulating the mitochondrial network and reprogramming glucose metabolism. Targeting Sp1 could be an effective strategy for the treatment of cervical cancer.
Assuntos
MicroRNAs , Neoplasias do Colo do Útero , Feminino , Humanos , Neoplasias do Colo do Útero/patologia , MicroRNAs/metabolismo , Transformação Celular Neoplásica , Glucose/metabolismo , Proliferação de Células , Fator de Transcrição Sp1/genética , Fator de Transcrição Sp1/metabolismo , Regulação Neoplásica da Expressão Gênica , Linhagem Celular TumoralRESUMO
Soluble fibrinogen-like protein 2 (sFgl2), a novel effector of regulatory T cells (Tregs), has been demonstrated to have potent immunosuppressive functions. Multiple studies indicate that Tregs could exert important atheroprotective effects, but their numbers gradually decrease during atherogenesis. The receptor of sFgl2 can be expressed on Treg precursor cells, while the role of sFgl2 on Treg differentiation and atherosclerosis progression remains unclear. Firstly, we detected that the sFgl2 was decreased in humans and mice with atherosclerotic diseases and was especially lower in their vulnerable plaques. Then, we used both Adeno-associated virus-sFgl2 (AAV-sFgl2)-injected ApoE-/- mice, which is systemic overexpression of sFgl2, and sFgl2TgApoE-/- bone marrow cells (BMC)-transplanted ApoE-/- mice, which is almost immune-system-specific overexpression of sFgl2, to explore the role of sFgl2 in atherosclerosis. Our experiment data showed that AAV-sFgl2 and BMT-sFgl2 could reduce atherosclerotic area and enhance plaque stability. Mechanistically, sFgl2 increases the abundance and immunosuppressive function of Tregs, which is partly mediated by binding to FcγRIIB receptors and phosphorylating Smad2/3. Collectively, sFgl2 has an atheroprotective effect that is mainly achieved by forming a positive feedback pathway with Treg. sFgl2 and Treg could synergistically protect against atherosclerosis.
Assuntos
Aterosclerose , Placa Aterosclerótica , Humanos , Animais , Camundongos , Linfócitos T Reguladores/metabolismo , Retroalimentação , Aterosclerose/genética , Aterosclerose/prevenção & controle , Aterosclerose/metabolismo , Placa Aterosclerótica/genética , Placa Aterosclerótica/metabolismo , Fibrinogênio/metabolismo , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismoRESUMO
Species of the genus Citrus are cultivated in many regions of China and are widely used for medicinal purposes. In the present study, essential oils (EOs) were extracted from four different Citrus species using steam distillation. The chemical components of these four essential oils were separated using gas chromatography-mass spectrometry, and 52 compounds were confirmed. D-limonene was found to be the most abundant compound. All four essential oils demonstrated varied but remarkable radical scavenging capacity (IC50 ; 0.77-13.9 %). Citrus paradisi essential oil exhibited excellent antioxidant activity. Compared to ibuprofen, topical application of the four Citrus spp. essential oils significantly inhibited ear edema formation in mice. Furthermore, essential oils from the four Citrus species reduced the expression levels of interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and nuclear transcription factor kappa B p65 (NF-κB) to different degrees. The cytotoxicity of the four essential oils on BV2 microglial cells was determined using the MTT assay (IC50 ; 321.37-1558.87â µg/mL), wherein Citrus limon essential oil showed the lowest cytotoxicity. The essential oils of Citrus limon, Citrus reticulata, and Citrus paradisi had an inhibitory effect on the lung cancer cell lines H1299 by inducing a G0/G1 cell cycle arrest. Cluster and principal component analyses were used to determine the relationship among the Citrus species. These results suggest that the four Citrus essential oils have potential for use as active ingredients in functional foods or cosmeceutical products.
Assuntos
Citrus paradisi , Citrus sinensis , Citrus , Óleos Voláteis , Animais , Citrus/química , Limoneno , Camundongos , Óleos Voláteis/química , Óleos Voláteis/farmacologiaRESUMO
Curcuma longa L. is one of the traditional Chinese herbs in the list of medicinal and food homology. Aromatic-turmerone is the main ingredient in turmeric essential oil. The aim of the present study is to investigate the treatment of Aromatic-turmerone on DSS-included colitis and its regulatory effect on intestinal flora disorder. Male KM mice supplemented with different concentration of aromatic-turmerone and mesalazine are subjected to 2% DSS in drinking water to induce colitis. Colon and cecum contents were collected for colitis lesion evaluation and inflammation-related gene analysis and colon contents for gut microbiota. The results show that treatments with Aromatic-turmerone significantly prevents colon shortening, alleviates the damage of colonic tissue, and reduces colonic inflammatory cytokines TNF-α and COX-2. Furthermore, the 16S rDNA gene sequence data indicate that Aromatic-turmerone improve the abundance of bacterial species, maintain some beneficial bacteria, and reduce harmful bacteria. Aromatic-turmerone downregulates the colonic inflammatory cytokines and modulates the abundance of intestinal flora, which is conductive to ameliorates DSS-induced colitis. Regularly intake of the edible herb may be help to prevent ulcerative colitis-related diseases.
Assuntos
Colite Ulcerativa , Colite , Microbioma Gastrointestinal , Animais , Colite/induzido quimicamente , Colite/tratamento farmacológico , Colite/patologia , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/tratamento farmacológico , Colo , Citocinas , Sulfato de Dextrana/farmacologia , Modelos Animais de Doenças , Cetonas , Masculino , Camundongos , Camundongos Endogâmicos C57BL , SesquiterpenosRESUMO
OBJECTIVE: Platelet activation by stimulatory factors leads to an increase in intracellular calcium concentration ([Ca2+]i), which is essential for almost all platelet functions. Modulation of Ca2+ influx and [Ca2+]i in platelets has been emerging as a possible strategy for preventing and treating platelet-dependent thrombosis. Voltage-gated potassium 1.3 channels (Kv1.3) are highly expressed in platelets and able to regulate agonist-evoked [Ca2+]i increase. However, the role of Kv1.3 channels in regulating platelet functions and thrombosis has not yet been elucidated. In addition, it is difficult to obtain a specific blocker for this channel, since Kv1.3 shares identical drug-binding sites with other K+ channels. Here, we investigate whether specific blockade of Kv1.3 channels by monoclonal antibodies affects platelet functions and thrombosis. Approach and Results: In this study, we produced the anti-Kv1.3 monoclonal antibody 6E12#15, which could specifically recognize both human and mouse Kv1.3 proteins and sufficiently block Kv1.3 channel currents. We found Kv1.3 blockade by 6E12#15 inhibited platelet aggregation, adhesion, and activation upon agonist stimulation. In vivo treatment with 6E12#15 alleviated thrombus formation in a mesenteric arteriole thrombosis mouse model and protected mice from collagen/epinephrine-induced pulmonary thromboembolism. Furthermore, we observed Kv1.3 regulated platelet functions by modulating Ca2+ influx and [Ca2+]i elevation, and that this is mediated in part by P2X1. Interestingly, Kv1.3-/- mice showed impaired platelet aggregation while displayed no abnormalities in in vivo thrombus formation. This phenomenon was related to more megakaryocytes and platelets produced in Kv1.3-/- mice compared with wild-type mice. CONCLUSIONS: We showed specific inhibition of Kv1.3 by the novel monoclonal antibody 6E12#15 suppressed platelet functions and platelet-dependent thrombosis through modulating platelet [Ca2+]i elevation. These results indicate that Kv1.3 could act as a promising therapeutic target for antiplatelet therapies.
Assuntos
Anticorpos Monoclonais/farmacologia , Plaquetas/efeitos dos fármacos , Fibrinolíticos/farmacologia , Canal de Potássio Kv1.3/antagonistas & inibidores , Inibidores da Agregação Plaquetária/farmacologia , Bloqueadores dos Canais de Potássio/farmacologia , Embolia Pulmonar/prevenção & controle , Trombose/prevenção & controle , Animais , Coagulação Sanguínea/efeitos dos fármacos , Plaquetas/metabolismo , Modelos Animais de Doenças , Feminino , Células HEK293 , Humanos , Canal de Potássio Kv1.3/sangue , Canal de Potássio Kv1.3/deficiência , Canal de Potássio Kv1.3/genética , Masculino , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ativação Plaquetária/efeitos dos fármacos , Embolia Pulmonar/sangue , Embolia Pulmonar/genética , Embolia Pulmonar/metabolismo , Transdução de Sinais , Trombose/sangue , Trombose/genética , Trombose/metabolismoRESUMO
Myoblast differentiation is a crucial process for myogenesis. Mitochondria function as an energy-providing machine that is critical to this process, and mitochondrial dysfunction can prevent myoblasts from fusing into myotubes. However, the molecular mechanisms underlying the dynamic regulation of mitochondrial networks remain poorly understood. In the present study, we found that the PTEN induced kinase 1 (PINK1)/Parkin (an E3 ubiquitin-protein ligase) pathway is activated at the early stage of myoblast differentiation. Moreover, downregulation of mitofusin 2 (Mfn2) and increased dynamin-related protein 1 (Drp1) resulted in loosely formed mitochondria during this period. Furthermore, selective knockdown of the mitochondrial matrix protein Lon peptidase-1 (LonP1) at the early stage of myoblast differentiation induced mitochondrial depolarization and suppressed the PINK1/Parkin pathway and reduced Mfn2 and Drp1 levels, which blocked mitochondrial remodeling and myoblast differentiation. Overall, these data demonstrate that LonP1 promotes myoblast differentiation by regulating PINK1/Parkin-mediated mitochondrial remodeling.
Assuntos
Proteases Dependentes de ATP/metabolismo , Diferenciação Celular/fisiologia , Mitocôndrias/metabolismo , Proteínas Mitocondriais/metabolismo , Mioblastos/metabolismo , Proteínas Quinases/metabolismo , Transdução de Sinais/fisiologia , Ubiquitina-Proteína Ligases/metabolismo , Animais , Camundongos , Proteínas Associadas aos Microtúbulos/metabolismo , Desenvolvimento Muscular/fisiologiaRESUMO
As a chronic musculoskeletal degeneration disease, intervertebral disc degeneration (IVDD) has been identified as a crucial cause for low back pain. This condition has a prevalence of 80% among adults without effective preventative therapy. Procyanidin B3 (Pro-B3) is a procyanidin dimer, which is widely present in the human diet and has multiple functions, such as preventing inflammation. But the inhibiting effect of Pro-B3 in IVDD development is still no known. Thus, our study aimed to demonstrate the therapeutical effect of Pro-B3 in IVDD and explain the underlying mechanism. In vitro studies, human nucleus pulposus (NP) cells were isolated and exposed in lipopolysaccharide (LPS) to simulate IVDD development. Pro-B3 pre-treatment inhibited LPS-induced production of inflammation correlated factors such as tumour necrosis factor α (TNF-α), interleukin-6 (IL-6), prostaglandin E2 (PGE2) and Nitric oxide (NO). On the other hand, LPS-medicated extracellular matrix (ECM) breakdown was blocked in Pro-B3 treated NP cells. Additionally, Pro-B3 treatment blocked the activation of NF-κB/toll-like receptor 4 pathway in LPS-exposed NP cells. Mechanistically, Pro-B3 could occupy MD-2's hydrophobic pocket exhibiting high affinity for LPS to intervene LPS/TLR4/MD-2 complex formation. In vivo, Pro-B3 treatment prevented the loss of gelatin NP cells and structural damage of annulus fibrosus in rat IVDD model. In brief, Pro-B3 is considered to be a treatment agent for IVDD.
Assuntos
Biflavonoides/uso terapêutico , Catequina/uso terapêutico , Degeneração do Disco Intervertebral/tratamento farmacológico , Degeneração do Disco Intervertebral/metabolismo , Antígeno 96 de Linfócito/metabolismo , Proantocianidinas/uso terapêutico , Receptor 4 Toll-Like/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Biflavonoides/química , Biflavonoides/farmacologia , Catequina/química , Catequina/farmacologia , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Progressão da Doença , Matriz Extracelular/metabolismo , Humanos , Degeneração do Disco Intervertebral/patologia , Lipopolissacarídeos , Masculino , Fator 88 de Diferenciação Mieloide/metabolismo , NF-kappa B/metabolismo , Núcleo Pulposo/patologia , Proantocianidinas/química , Proantocianidinas/farmacologia , Punções , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Fator 6 Associado a Receptor de TNF/metabolismoRESUMO
Melittin (MEL), the primary active component of bee venom, has recently emerged as a promising cancer chemotherapeutic agent. However, the instability and rapid degradation of MEL is a significant challenge in practical therapeutic applications. In the present study, graphene oxide (GO)-based magnetic nanocomposites (PEG-GO-Fe3O4) were prepared and adopted as the drug delivery vehicles of MEL, and the anticancer effects of PEG-GO-Fe3O4/MEL complexes on human cervical cancer HeLa cells were studied. PEG-GO-Fe3O4 exhibited a series of unique physical and chemical properties resulting in multiple interactions with MEL, and ultimately the release of MEL. In vitro experiments showed that PEG-GO-Fe3O4/MEL not only distinctly enhanced the inhibition effect on HeLa cells, but also induced pore formation in the cell membrane that ultimately led to cell lysis. In this newly developed drug delivery system, PEGylated GO plays the role of a MEL protector while Fe3O4 nanoparticles act as magnetic responders; therefore active MEL can be released over a long period of time (up to 72 h) and maintain its inhibition effect on HeLa cells.
Assuntos
Grafite/química , Meliteno/farmacologia , Neoplasias do Colo do Útero/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sistemas de Liberação de Medicamentos , Estabilidade de Medicamentos , Feminino , Células HeLa , Humanos , Meliteno/química , Nanocompostos , Polietilenoglicóis/químicaRESUMO
Previous work has confirmed that the chronic hypoxia-hypercapnia (CHH) associated with chronic obstructive pulmonary disease contributes to the development of skeletal muscle atrophy. Neuromuscular Electrical Stimulation (NMES) has shown some efficacy when used as a treatment to reduce skeletal muscle atrophy. The present study focuses on the MicroRNA-486/PTEN/FoxO1 pathway with the goal of identifying its physiological role in skeletal muscle atrophy induced by CHH as well as its role during NMES treatment. To test this, 32 male Sprague Dawley rats were randomly divided into four groups. After completion of the disease modeling, gastrocnemius muscles were collected from all animals and cross-sectional areas of muscular fiber were observed and analyzed via H&E staining. MiR-486 expression was further assessed by qRT-PCR, and protein levels of TNF-α, PTEN, p-Akt, Akt, FoxO1, atrogin-1 and MuRF1 were measured by immunohistochemistry and western blotting. CSA, miR-486, and the ratio p-Akt/Akt were significantly reduced in the CHH group, while the levels of TNF-α, PTEN, FoxO1, atrogin-1, and MuRF1 were markedly increased. Importantly, these findings were reversed as a result of NMES. Thus, the MicroRNA-486/PTEN/FoxO1 pathway functions in muscle protein synthesis and degradation. NEW & NOTEWORTHY: Our research provides a theoretical basis for the application of NMES as a means of improving muscle atrophy. Moreover, these therapeutic targets provide possible clues relevant to the treatment of amyotrophic diseases.
Assuntos
Estimulação Elétrica , Hipercapnia/complicações , Hipóxia/complicações , MicroRNAs/metabolismo , Atrofia Muscular/terapia , Proteínas do Tecido Nervoso/metabolismo , Animais , Hipercapnia/fisiopatologia , Hipóxia/fisiopatologia , Proteínas Musculares/biossíntese , Proteínas Musculares/metabolismo , Atrofia Muscular/etiologia , PTEN Fosfo-Hidrolase/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Yes-associated protein (Yap) is a core transcriptional coactivator in the downstream Hippo pathway that regulates cell proliferation and tissue growth. However, its role in the regulation of myoblast differentiation remains unclear. Regulation of mitochondrial networks by dynamin-related protein 1 (Drp1) and mitofusion 2 (Mfn2) is crucial for the activation of myoblast differentiation. In the present study, we investigated the interplay between the Hippo/Yap pathway and protein contents of Mfn2 and Drp1 during myoblast differentiation. The Hippo/Yap pathway was inactivated at the early stage of myoblast differentiation due to the decreased ratio of phosphorylated mammalian sterile 20 kinases 1/2 (p-Mst1/2) to Mst1/2, phosphorylated large tumor suppressor 1 (p-Lats1) to Lats1, and phosphorylated Yap (serine 112, p-Yap S112) to Yap, which resulted in the translocation of Yap from cytoplasm to nucleus, increased protein content of Drp1, and mitochondrial fission events. Downregulation of Yap inhibited myoblast differentiation and decreased the content of Drp1, which resulted in elongated mitochondria, fused mitochondrial networks, and collapsed mitochondrial membrane potential. Together, our data indicate that inactivation of the Hippo/Yap pathway could induce mitochondrial fission by promoting Drp1 content at the early stage of myoblast differentiation.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Diferenciação Celular/fisiologia , Mitocôndrias/metabolismo , Mitocôndrias/fisiologia , Mioblastos/metabolismo , Fosfoproteínas/metabolismo , Animais , Proteínas de Ciclo Celular , Regulação para Baixo/fisiologia , Dinaminas/metabolismo , GTP Fosfo-Hidrolases/metabolismo , Potencial da Membrana Mitocondrial/fisiologia , Camundongos , Dinâmica Mitocondrial/fisiologia , Mioblastos/fisiologia , Fosforilação/fisiologia , Transdução de Sinais/fisiologia , Proteínas de Sinalização YAPRESUMO
Previous studies have confirmed that selective blockade of Kv1.3 channels could modulate the activities of pathogenic T cells and microglia/macrophages, which play key roles in experimental autoimmune encephalomyelitis (EAE). In this study, we designed an anti-Kv1.3 vaccine (PADRE-Kv1.3) to explore its protective role in EAE rat models. When the vaccine was applied in EAE rats, clinical scores and several staining techniques were used to evaluate the severity of the disease. T cell subtypes and related cytokines, as well as microglia/macrophage activation were assayed through flow cytometry, qRT-PCR or immunofluorescence staining, respectively. We herein showed that rats and mice developed high titers of anti-Kv1.3 antibodies and appeared no abnormal manifestations after the PADRE-Kv1.3 vaccine treatment. In EAE models, the vaccine treatment effectively alleviated the clinical severity and lessened pathological damages in the central nervous system (CNS). In addition, we found the vaccine significantly decreased the number of pathogenic T cells (Th17 and IFN-γ-producing T cells) and the production of related pro-inflammatory cytokines (IL-17A, IFN-γ and IL-1ß), but increased the number of protective T subsets (CD4+IL-10+ T cells and Treg cells) in the spleen or CNS. Moreover, the infiltration of microglia/macrophages significantly reduced and these cells shifted toward anti-inflammatory M2 subtype in the CNS after the vaccine treatment. Thus, we demonstrated that the PADRE-Kv1.3 vaccine could induce therapeutic anti-Kv1.3 antibodies and ameliorate EAE in rats effectively and safely, which provides a new field of vision for the protection and therapy of multiple sclerosis.
Assuntos
Sistema Nervoso Central/imunologia , Encefalomielite Autoimune Experimental/imunologia , Canal de Potássio Kv1.3/metabolismo , Macrófagos/imunologia , Microglia/imunologia , Esclerose Múltipla/imunologia , Linfócitos T Reguladores/imunologia , Células Th17/imunologia , Vacinas de Subunidades Antigênicas/imunologia , Animais , Células Cultivadas , Sistema Nervoso Central/patologia , Citocinas/metabolismo , Modelos Animais de Doenças , Epitopos de Linfócito B/genética , Cobaias , Humanos , Mediadores da Inflamação/metabolismo , Canal de Potássio Kv1.3/genética , Vacinas Antimaláricas/genética , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ratos , Ratos Endogâmicos Lew , Proteínas Recombinantes de Fusão/genética , Vacinas de Subunidades Antigênicas/genéticaRESUMO
Numerous evidence suggests that RhoA/Rho kinase (ROCK) signaling pathway plays an important role in the pathogenesis of pulmonary arterial hypertension (PAH), but little is known about its effects on the development of PAH in mice with absence of the adenosine A2A receptor (A2AR). Eight A2AR knockout (KO) and 8 wild-type mice were used. Morphometric analysis of pulmonary arterioles included right ventricle/left ventricle plus ventricular septum (Fulton index), vessel wall thickness/total vascular diameter (WT%), and vessel wall area/total vascular area (WA%). The expression of RhoA and ROCK1 mRNA was determined by real-time polymerase chain reaction. The expression of RhoA, ROCK1, and phosphorylation of myosin phosphatase target subunit 1 proteins in pulmonary tissue was tested by Western blot. The position of ROCK1 protein was evaluated by immunohistochemistry. Compared with wild-type mice, A2AR KO mice displayed (1) increased Fulton index, WT%, and WA% (P < 0.01); (2) increased mRNA expression of RhoA and ROCK1 (each P < 0.05); (3) increased protein expression of RhoA, ROCK1, and phosphorylation of myosin phosphatase target subunit 1 (each P < 0.01); (4) increased location of ROCK1 protein in endothelial and smooth muscle cells of pulmonary artery, bronchial, and alveolar epithelial cells. Activation of RhoA/ROCK signaling pathway may cause pulmonary vascular constriction, pulmonary artery remodeling, and PAH in adenosine A2A receptor KO mice.
Assuntos
Hipertensão Pulmonar/metabolismo , Receptor A2A de Adenosina/deficiência , Transdução de Sinais/fisiologia , Proteínas rho de Ligação ao GTP/metabolismo , Quinases Associadas a rho/metabolismo , Animais , Hipertensão Pulmonar/patologia , Masculino , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteína rhoA de Ligação ao GTPRESUMO
AIMS: γ-aminobutyric acid (GABA), the principal inhibitory neurotransmitter, acts on GABA receptors to play an important role in the modulation of macrophage functions. The present study examined the effects of GABA and a GABA receptor agonist on modulating cholesterol-metabolism-associated molecules in human monocyte-derived macrophages (HMDMs). METHODS: ORO stain, HPLC, qRT-PCR, Western blot and EMSA were carried out using HMDMs exposed to ox-LDL with or without GABAergic agents as the experimental model. RESULTS: GABA and topiramate reduced the percentage of cholesterol ester in lipid-laden HMDMs by down-regulating SR-A, CD36 and LOX-1 expression and up-regulating ABCA1, ABCG1 and SR-BI expression in lipid-laden HMDMs. The production of TNF-α was decreased in GABA-and topiramate-treated lipid-laden HMDMs, and levels of interleukin (IL)-6 did not change. The activation of two signaling pathways, p38MAPK and NF-κB, was repressed by GABA and topiramate in lipid-laden HMDMs. CONCLUSION: GABA and topiramate inhibit the formation of human macrophage-derived foam cells and may be a possibility for macrophage targeted therapy of atherosclerotic lesions.
Assuntos
Colesterol/metabolismo , Células Espumosas/efeitos dos fármacos , Frutose/análogos & derivados , Fármacos Neuroprotetores/farmacologia , Ácido gama-Aminobutírico/farmacologia , Transportador 1 de Cassete de Ligação de ATP/genética , Transportador 1 de Cassete de Ligação de ATP/metabolismo , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Antígenos CD36/genética , Antígenos CD36/metabolismo , Regulação para Baixo/efeitos dos fármacos , Células Espumosas/citologia , Frutose/farmacologia , Humanos , Lipoproteínas LDL/farmacologia , Macrófagos/citologia , Fosforilação/efeitos dos fármacos , Receptores Depuradores Classe A/genética , Receptores Depuradores Classe A/metabolismo , Receptores Depuradores Classe E/genética , Receptores Depuradores Classe E/metabolismo , Transdução de Sinais , Topiramato , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Background: Bone tissue engineering (BTE) is a promising alternative to autologous bone grafting for the clinical treatment of bone defects, and inorganic/organic composite hydrogels as BTE scaffolds are a hot spot in current research. The construction of nano-hydroxyapatite/gelatin methacrylate/oxidized sodium alginate (nHAP/GelMA/OSA), abbreviated as HGO, composite hydrogels loaded with bone morphogenetic protein 7 (BMP7) will provide a suitable 3D microenvironment to promote cell aggregation, proliferation, and differentiation, thus facilitating bone repair and regeneration. Methods: Dually-crosslinked hydrogels were fabricated by combining GelMA and OSA, while HGO hydrogels were formulated by incorporating varying amounts of nHAP. The hydrogels were physically and chemically characterized followed by the assessment of their biocompatibility. BMP7-HGO (BHGO) hydrogels were fabricated by incorporating suitable concentrations of BMP7 into HGO hydrogels. The osteogenic potential of BHGO hydrogels was then validated through in vitro experiments and using rat femoral defect models. Results: The addition of nHAP significantly improved the physical properties of the hydrogel, and the composite hydrogel with 10% nHAP demonstrated the best overall performance among all groups. The selected concentration of HGO hydrogel served as a carrier for BMP7 loading and was evaluated for its osteogenic potential both in vivo and in vitro. The BHGO hydrogel demonstrated superior in vitro osteogenic induction and in vivo potential for repairing bone tissue compared to the outcomes observed in the blank control, BMP7, and HGO groups. Conclusion: Using hydrogel containing 10% HGO appears promising for bone tissue engineering scaffolds, especially when loaded with BMP7 to boost its osteogenic potential. However, further investigation is needed to optimize the GelMA, OSA, and nHAP ratios, along with the BMP7 concentration, to maximize the osteogenic potential.
Assuntos
Alginatos , Proteína Morfogenética Óssea 7 , Regeneração Óssea , Durapatita , Gelatina , Hidrogéis , Osteogênese , Engenharia Tecidual , Alicerces Teciduais , Alginatos/química , Alginatos/farmacologia , Animais , Proteína Morfogenética Óssea 7/química , Proteína Morfogenética Óssea 7/farmacologia , Gelatina/química , Engenharia Tecidual/métodos , Hidrogéis/química , Hidrogéis/farmacologia , Durapatita/química , Durapatita/farmacologia , Osteogênese/efeitos dos fármacos , Ratos , Regeneração Óssea/efeitos dos fármacos , Alicerces Teciduais/química , Ratos Sprague-Dawley , Metacrilatos/química , Masculino , Humanos , Osso e Ossos/efeitos dos fármacosRESUMO
OBJECTIVE: CT scans combined with Mimics software were used to measure femoral offset (FO), rotation center height (RCH) and lower leg length discrepancy (LLD) following total hip arthroplasty (THA), and the relationship between FO, RCH and LLD after THA is discussed. METHODS: Retrospective analysis was performed on 40 patients with unilateral THA who met standard cases from October 2020 to June 2022. There were 21 males and 19 females, 18 patients on the left side and 22 patients on the right side, aged range from 30 to 81 years old, with an average age of (58.90 ±14.13) years old, BMI ranged from 17.3 to 31.5 kg·m-2 with an average of (25.3±3.4) kg·m-2. There were 30 cases of femoral head necrosis (Ficat type â £), 2 cases of hip osteoarthritis (Tönnis type â ¢), 2 cases of developmental hip dislocation combined with end-stage osteoarthritis (Crowe type â ¢), and 6 cases of femoral neck fracture (Garden type â £). Three-dimensional CT reconstruction of pelvis was taken preoperative and postoperative, and three-dimensional reconstruction model was established after processing by Mimics software. FO, RCH and LLD were measured on the model. The criteria for FO reconstruction were as follows:postoperative bilateral FO difference less than 5 mm;the standard for equal length of both lower limbs was as follows:postoperative LLD difference less than 5 mm. RESULTS: Bilateral FO difference was positively correlated with LLD (r=0.744, P<0.001). Chi-square test was performed between the FO reconstructed group and the non-reconstructed eccentricity group:The results showed that the isometric ratio of lower limbs in the FO reconstructed group was significantly higher than that in the FO reconstructed group (χ2=6.320, P=0.012). The bilateral RCH difference was significantly negatively correlated with LLD(r=-0.877, P<0.001). There is a linear relationship between bilateral FO difference and bilateral RCH difference and postoperative LLD, and the linear regression equation is satisfied:postoperative LLD=0.038x-0.099y+0.257(x:postoperative bilateral FO difference, y:postoperative bilateral RCH difference; Unit:cm), F=77.993, R2=0.808, P=0.009. CONCLUSION: After THA, LLD increased with the increase of FO and decreased with the increase of RCH. The effect of lower limb isometric length can be obtained more easily by reconstruction of FO. There is a linear relationship between the bilateral FO difference and the bilateral RCH difference after THA and LLD, and the regression equation can provide a theoretical reference for judging LLD.
Assuntos
Artroplastia de Quadril , Fêmur , Desigualdade de Membros Inferiores , Humanos , Masculino , Feminino , Desigualdade de Membros Inferiores/etiologia , Idoso , Pessoa de Meia-Idade , Artroplastia de Quadril/métodos , Idoso de 80 Anos ou mais , Estudos Retrospectivos , Adulto , Fêmur/cirurgia , Tomografia Computadorizada por Raios X , Rotação , Osteoartrite do Quadril/cirurgia , Osteoartrite do Quadril/etiologiaRESUMO
Using DIVA-GIS software to study the spatial accumulation of Citrus species, an important economic crop in China. Draw the distribution maps of Citrus to concerning altitude and vegetation, and use DIVA-GIS' Bioclim ecological model and maximum information entropy model Maxent software to estimate the potential distribution areas of various Citrus species. The results show that the Citrus genus is located in the south of Qinling Mountains, mainly in the southwest of China and the coast of southeastern China. Sichuan and Chongqing are the most densely distributed regions of Citrus. The distribution of Citrus is closely related to the vegetation type and altitude. The vegetation types in the distribution area is evergreen broad-leaved forest, evergreen shrub, deciduous broad-leaved forest, mixed-leaf forest coverage area, deciduous coniferous forest, farmland, trees, other vegetation coverage, and evergreen coniferous forest. The current potential distribution area of Citrus is mainly in Guizhou, Hubei, Hunan, Guangdong, Guangxi, Yunnan, Sichuan, Chongqing, and other provinces and municipalities and their borders, while the potential distribution area in the future moves northward and enter Henan and southern Gansu. At the same time, climate warming changes the distribution of suitable areas of Citrus, which makes the suitable areas of C. sinensis Osbeck, C. reticulata Blanco, and other crops greatly increased. Planning the planting area will effectively improve the yield and quality in the future. Planning presents new challenges.
Assuntos
Clima , Árvores , China , Florestas , AltitudeRESUMO
Climate change is closely related to the distribution of plant resources. Cerasus serrulata is an important plant resource in China. The study on the influence of environmental factors on the distribution of suitable areas of C. serrulata is conducive to the protection and development of C. serrulata. In this paper, the distribution information of 238 Chinese Cerasus serrulata plants was processed by DIVA-GIS. The MaxEnt model was used to simulate the current and future distribution, and the ecological distribution and richness of Cerasus were analyzed. The results showed that the Cerasus serrulata was widely distributed in Hebei, Heilongjiang, Shandong, Jiangsu, Zhejiang, Anhui, Jiangxi, Hunan, and Guizhou provinces, mainly in the low and middle elevation areas of 10 to 1200 m. Based on this model, the precipitation of the warmest quarter, the precipitation of the driest month, and the mean temperature of the coldest were the most significant bioclimatic variables affecting the distribution of C. serrulata. In the future, climate change may lead to a slight increase of 2.31% in the area of suitable habitat for Cerasus serrulata, while the optimal habitat will decrease from 20.81 to 14.55%. Therefore, conservation measures should be taken to protect these precious resources.