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1.
Biochem Biophys Res Commun ; 694: 149409, 2024 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-38141558

RESUMO

The NLRP3 inflammasome has gained significant attention due to its participation in diverse cellular processes. Nevertheless, the detailed framework of the canonical NLRP3 inflammasome assembly still remains unrevealed. This study aims to elucidate the transcriptomic landscape of the various stages involved in the canonical activation of the NLRP3 inflammasome in BMDMs by integrating RNA-seq, bioinformatics, and molecular dynamics analyses. The model for the canonical activation of the NLRP3 inflammasome was confirmed through morphological observations, functional assessments (ELISA and LDH), and protein detection (western blot). Subsequently, cells were subjected to RNA sequencing following three groups: control, priming (LPS 500 ng/ml, 4 h), and activation (LPS 500 ng/ml, 4 h; ATP 5 mM, 1 h). A total of 9116 differentially expressed genes (DEGs) were identified, which exerted regulatory effects on various pathways, including cell metabolism, ion fluxes, post-translational modifications, and organelles. Subsequently, six hub genes (Sirt3, Stat3, Syk, Trpm2, Tspo, and Txnip) were identified via integrating literature review and database screening. Finally, the three-dimensional structures of these six hub proteins were obtained using the MD-optimized RoseTTAFold and Gromacs simulations (at least 200 ns). In summary, our research offers novel insights into the transcriptomic-level understanding of the assembly of the canonical NLRP3 inflammasome.


Assuntos
Inflamassomos , Proteína 3 que Contém Domínio de Pirina da Família NLR , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Lipopolissacarídeos/metabolismo , Macrófagos/metabolismo , Perfilação da Expressão Gênica , Interleucina-1beta/metabolismo
2.
Fish Shellfish Immunol ; 145: 109322, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38128679

RESUMO

Nocardia seriolae is a severe bacterial pathogen that has seriously affected the development of aquaculture industry. Largemouth bass (Micropterus salmoides) is a commercially significant freshwater fish that suffers a variety of environmental threats, including bacterial pathogens. However, the immune responses and metabolic alterations of largemouth bass to N. seriolae infection remain largely unclear. We discovered that N. seriolae caused pathological alterations in largemouth bass and shifted the transcript of immune-related and apoptotic genes in head kidney after infection. To answer the aforementioned question, a combined transcriptome and metabolome analysis was employed to explore the alterations in genes, metabolites, and metabolic pathways in largemouth bass following bacterial infection. A total of 3579 genes and 1929 metabolites are significant differentially changed in the head kidney post infection. In response to N. seriolae infection, host modifies the PI3K-Akt signaling pathway, TCA cycle, glycolysis, and amino acid metabolism. The integrated analysis of transcriptome and metabolome suggested that with the arginine metabolism pathway as the core, multiple biomarkers (arg gene, arginine) are involved in the antibacterial and immune functions of largemouth bass. Thus, we hypothesized that arginine plays a crucial role in the immune responses of largemouth bass against N. seriolae infection, and increasing arginine levels suitably is beneficial for the host against bacterial infection. Our results shed light on the regulatory mechanism of largemouth bass resistance to N. seriolae infection and contributed to the development of more effective N. seriolae resistance strategies.


Assuntos
Infecções Bacterianas , Bass , Nocardiose , Nocardia , Animais , Transcriptoma , Fosfatidilinositol 3-Quinases/genética , Metaboloma , Arginina
3.
J Environ Manage ; 359: 120914, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38669886

RESUMO

RESEARCH QUESTION: In the context of global low-carbon emission reduction, how to achieve green and high-quality development has become a major issue for the Communist Party of China (CPC) and the Chinese government recently. Based on the data of China's listed companies from 2013 to 2020, this paper uses Python to implement text analysis of annual reports, and explores the relationship between political acuity and carbon-efficiency synergies (CES) from the perspective of enterprise initiative. RESEARCH FINDINGS: We found that (1) political acuity positively affects carbon-efficiency synergies. (2) Increased political acuity can reduce carbon emissions, but the effect on economic efficiency is not obvious. That is, low carbon takes the lead in raising the level of carbon-efficiency synergies. (3) Environmental regulations can positively regulate the relationship between political acuity and carbon-efficiency synergies. (4) Political acuity in southern China, carbon neutral and non-state-owned enterprises (NSOEs) will have a more pronounced effect on carbon-efficiency synergies. ACADEMIC IMPLICATIONS: From the perspective of the root causes of political linkages, we find the synergies between formal and informal institutions, and the key factors for policy implementation. POLICY IMPLICATIONS: This paper is helpful for enterprises to improve the synergies of emission reduction and efficiency promotion, and has practical implications for the government to promote green and high-quality development.


Assuntos
Carbono , China , Política , Política Ambiental
4.
Fish Shellfish Immunol ; 122: 131-145, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35122948

RESUMO

Haliotis discus hannai is a commercially important mollusk species, and the abalone aquaculture sector has been jeopardized by deteriorating environmental circumstances such as bacterial infection and thermal stress during the hot summers. However, due to a paucity of genetic information, such as transcriptome resources, our understanding of their stress adaptation is restricted. In this research, using single-molecule long-read (SMRT) sequencing technology, a library composed of ten tissues (i.e., haemocytes, gills, muscle, hepatopancreas, digestive tract, mantle, mucous gland, ovary, testis and head) was constructed and sequenced. In all, 41,855 high-quality unique transcripts, among which 24,778 were successfully annotated. Additionally, 13,463 SSRs, 1,169 transcription factors, and 18,124 lncRNAs were identified in H. discus hannai transcriptome. Furthermore, multiple immune-related transcripts were identified according to KEGG annotation, and a portion of these transcripts were mapped into several classical immune-related pathways, including the PI3K-AKT signaling pathway and Toll-like receptor signaling pathway. Additionally, 24 typical sequences related to the immunity pathway were detected by RT-PCR; the results showed that most of the immune-related genes showed significantly high expression at 72 h after bacterial challenges and thermal stress, especially the expression level of genes in gills was significantly higher than that in haemocytes under V. parahaemolyticus stress at 24 h. At the same time. The analysis of alternative splicing identified several innate immunity-related functions genes, including CD109 and caspase 2. These results suggest that the complex immune system, particularly the powerful innate immunity system, was crucial for H. discus hannai response to numerous environmental challenges.


Assuntos
Gastrópodes , Fosfatidilinositol 3-Quinases , Animais , Sequência de Bases , Feminino , Gastrópodes/genética , Imunidade Inata/genética , Fosfatidilinositol 3-Quinases/genética , Análise de Sequência de RNA
5.
Fish Shellfish Immunol ; 125: 128-140, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35523358

RESUMO

Traditionally, invertebrates were thought to lack immune memory owing to a lack of acquired immune-related factors such as immunoglobulin. Nonetheless, with the in-depth consideration of invertebrate immune priming, scholars have gradually realized that the immune defenses of invertebrates are more complex than previously imagined. In the current investigation, the survival rate of Vibrio parahaemolyticus re-infected Haliotis discus hannai (VV group) was significantly different from the other groups (p < 0.05), indicating that an enhanced immune response may commence after first exposure to the same strain of V. parahaemolyticus. The transcriptome profiles of hemocytes obtained 102,052 unigenes, and 27,449 of them were annotated successfully. Venn diagram analysis showed that 2832 DEGs commonly responded to the first and second immune responses. 1734 "immune response genes" and 1460 "potential immune-enhancing genes" were also identified. A comparison of both "immune response genes" and "potential immune-enhancing genes" revealed 1019 immune-enhancing regulatory genes and 281 essential immune-enhancing genes. According to the KEGG enrichment analysis results of ERGs and EEGs, classical immune-related signaling pathways, such as NF-kappa B signaling pathway, NOD-like receptor signaling pathway, IL-17 signaling pathway, and TLR signaling pathway were significantly enriched, indicating that they were all involved in the response to V. parahaemolyticus re-infection and were likely dominant in the immune enhancement process of H. discus hannai hemocytes. The intermolecular interactions generated by Cytoscape after re-infection of V. parahaemolyticus appear more intuitively to demonstrate that hemocytes regulation was not an independent process, but rather an intricate regulatory network. H. discus hannai demonstrated enhanced immunological activity after re-infection with V. parahaemolyticus, showing immune memory in hemocytes. The current study's findings have broadened the study of immune enhancement in invertebrates and laid the framework for future research into the molecular mechanism of immune enhancement in abalones.


Assuntos
Gastrópodes , Vibrio parahaemolyticus , Animais , Gastrópodes/genética , Imunidade Inata/genética , Reinfecção , Transcriptoma , Vibrio parahaemolyticus/fisiologia
6.
Fish Shellfish Immunol ; 124: 300-312, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35398223

RESUMO

Long non-coding RNAs (lncRNAs) have been linked to immunological modulation. Unfortunately, little is known about the processes of immune control in shrimp. In crustaceans such as Litopenaeus vannamei, a prominent aquaculture species, the X-organ-sinus gland complex (XO-SG) in the eyestalk is an essential neuroendocrine regulatory organ. Eyestalk ablation is commonly employed in aquaculture to accelerate ovarian maturation in shrimp. It does, however, have a negative impact on the shrimps' immunocompetence and causes death. As a result, we used RNA-seq to profile the transcriptomes of L. vannamei hemocytes infected with Vibrio parahaemolyticus after the eyestalk ablation. Following strict transcript screening procedures, 2307 lncRNAs were identified from L. vannamei hemocytes in this study. Pearson correlation analysis was finally used to uncover 535 DElncRNAs and 1566 DEmRNA targets. According to the Venn diagram analysis, 326 non-eyestalk regulatory lncRNAs (NElncRNAs) with a target of 1014 non-eyestalk regulatory genes (NEmRNAs), 47 eyestalk negative regulatory lncRNAs (ENRlncRNAs) with a target of 95 eyestalk negative regulatory genes (ENRmRNAs), and 162 eyestalk positive regulatory lncRNAs (EPRlncRNAs) with a target of 457 eyestalk positive regulatory genes (EPRmRNAs) were screened. The bioinformatics analysis revealed that lncRNAs were associated with Axon regeneration, Rap1 signaling pathway, Thyroid hormone signaling pathway, TGF-beta signaling pathway, and PI3K-Akt signaling pathway, implying that lncRNAs may play a role in the regulation of the neuroendocrine-immune (NEI) system. Furthermore, several lncRNAs targeting HSP70, YWHAZ, FER2, HIF1α, and Notch were discovered and verified by qRT-PCR. These findings showed that regulation of lncRNAs in hemocytes which were controlled by the eyestalk might be one of the impact variables in controlling the differential expression of mRNAs associated with immune response in L. vannamei infected with V. parahaemolyticus.


Assuntos
Penaeidae , RNA Longo não Codificante , Vibrioses , Vibrio parahaemolyticus , Animais , Proteínas de Artrópodes , Axônios , Imunidade Inata/genética , Regeneração Nervosa , Fosfatidilinositol 3-Quinases/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Vibrio parahaemolyticus/genética
7.
Fish Shellfish Immunol ; 121: 62-73, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34998096

RESUMO

The X-organ-sinus gland complex (XO-SG) in the eyestalk is an important neuroendocrine regulatory organ of crustaceans such as Litopenaeus vannamei, a prominent aquaculture species. The current study found significant changes in the enzyme activities of ALP, ACP, and T-SOD of hepatopancreatic in response to Vibrio parahaemolyticus exposure following eyestalk ablation, indicating that they were all involved in the immunological regulation of shrimps against V. parahaemolyticus infection. A total of 52,656 unigenes were obtained after RNA-Seq, with an average length of 1036 bp and an N50 of 1847 bp. Subsequently, 1899 eyestalk positive regulation genes (EPRGs), 745 eyestalk negative regulation genes (ENRGs), and 2077 non-eyestalk regulatory genes (NEGs) were identified. KEGG analysis of EPRGs revealed that eyestalk ablation might activate the neuroendocrine-immune (NEI) system. The RNA-Seq data were validated using quantitative real-time PCR (qRT-PCR). The findings suggested that eyestalk ablation might affect the expression of genes involved in the prophenoloxidase-activating system, the TLR signaling pathway, and numerous other immune-related genes in L. vannamei. All of these findings revealed that the eyestalk might have a role in the immune response of L. vannamei. The genes and pathways discovered in this study will help to elucidate the molecular mechanisms of hemocytes' immune response to V. parahaemolyticus following eyestalk ablation in shrimp, as well as provide the framework for building crustacean immunity theory.


Assuntos
Estruturas Animais/imunologia , Imunidade Inata , Penaeidae , Vibrioses , Animais , Aquicultura , Hemócitos , Penaeidae/genética , Penaeidae/imunologia , Vibrio parahaemolyticus
8.
J Anat ; 239(5): 1134-1140, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34235729

RESUMO

The rat is frequently used as a model to study the characteristics, aetiology and pathology of the Achilles tendon. However, though the structure of the human Achilles tendon has been extensively investigated, the anatomical structure of the rat Achilles tendon remains unclear, which impedes the ability to use rats to study Achilles tendinopathy. The purpose of this study was to reveal the structure of the rat Achilles tendon and to explore its similarities with the human Achilles tendon through an anatomical dissection of 80 rat Achilles tendons (40 female, 40 male). This study found that the subtendons of the rat Achilles tendon originating from the triceps surae muscle were twisted, and each subtendon also had its own torsion. The extent of these two types of torsion could be very different between rats. Alterations in this torsion may result in distinct stress fields in the Achilles tendon, which may play a critical role in the pathogenesis of Achilles tendinopathy. This study provides an important basis to support the use of rats as model animals to investigate the characteristics of the human Achilles tendon and Achilles tendinopathy.


Assuntos
Tendão do Calcâneo , Tendinopatia , Animais , Dissecação , Feminino , Masculino , Músculo Esquelético , Ratos
9.
FASEB J ; 34(7): 9393-9404, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32474999

RESUMO

Parasitic infections are a severe issue in many regions of the world. We assume that if a chemical can destroy a DNA barcode sequence, then this chemical could be developed as a species-specific parasiticidal agent. To test this hypothesis, we designed sgRNAs that target the sequences of both a DNA barcode (ITS-2) and a control (5.8S rDNA) in Cryptocaryon irritans. In in vivo tests, we found that exposure to Cas9 mRNA mixed with sgRNAs was able to significantly reduce the hatching rate of tomont and the survival rate of theront. Quantitative Real-time PCR demonstrated that the DNAs of tomont and theront exposed to sgRNAs and Cas9 mRNA were significantly disrupted, no matter whether they were exposed to a single sgRNA or a mixture of two sgRNAs. DNA sequencing also suggested the test group that was exposed to a single sgRNA mixed with Cas9-induced mutation at sgRNA targeted fragments and the test group exposed to two sgRNAs combined with Cas9-induced deletion of large pieces. The findings and principles provided by this study contribute to the development of novel nucleic acid therapeutic drugs for cryptocaryoniasis and other parasitic diseases and provide insight into the development of species-specific parasiticidal agents.


Assuntos
Sistemas CRISPR-Cas , Infecções por Cilióforos/veterinária , Cilióforos/genética , Doenças dos Peixes/parasitologia , Edição de Genes , Proteínas de Protozoários/genética , RNA Guia de Cinetoplastídeos/genética , Animais , Cilióforos/crescimento & desenvolvimento , Cilióforos/metabolismo , Infecções por Cilióforos/parasitologia
10.
Int J Mol Sci ; 22(23)2021 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-34884814

RESUMO

Peritonitis caused by LPS is a severe clinical challenge, which causes organ damage and death. However, the mechanism of LPS-induced peritonitis has not been fully revealed yet. Here, we investigated the transcriptome profile of the peritoneal tissue of LPS-induced peritonitis in mice. A model of LPS-induced peritonitis in mice was established (LPS 10 mg/kg, i.p.), and the influence of TAK 242 (TLR4 inhibitor) on the level of inflammatory cytokines in mouse peritoneal lavage fluid was investigated by using an ELISA test. Next, the peritoneal tissues of the three groups of mice (Control, LPS, and LPS+TAK 242) (n = 6) were isolated and subjected to RNA-seq, followed by a series of bioinformatics analyses, including differentially expressed genes (DEGs), enrichment pathway, protein-protein interaction, and transcription factor pathway. Then, qPCR verified-hub genes that may interact with TAK 242 were obtained. Subsequently, the three-dimensional structure of hub proteins was obtained by using homology modeling and molecular dynamics optimization (300 ns). Finally, the virtual docking between TAK 242 and hub proteins was analyzed. Our results showed that TAK 242 significantly inhibited the production of inflammatory cytokines in the peritoneal lavage fluid of mice with peritonitis, including IL-6, IFN-γ, IL-1ß, NO, and TNF-α. Compared with the Control group, LPS treatment induced 4201 DEGs (2442 down-regulated DEGs and 1759 up-regulated DEGs). Compared with the LPS group, 30 DEGs were affected by TAK 242 (8 down-regulated DEGs and 22 up-regulated DEGs). A total of 10 TAK 242-triggered hub genes were obtained, and the possible docking modes between TAK 242 and hub proteins were acquired. Overall, our data demonstrated that a large number of DEGs were affected in LPS-triggered peritonitis mice. Moreover, the TLR4 inhibitor TAK 242 is capable of suppressing the inflammatory response of LPS-induced peritonitis. Our work provides clues for understanding the pathogenesis of LPS-induced peritonitis in mice.


Assuntos
Citocinas/análise , Lipopolissacarídeos/toxicidade , Peritônio/patologia , Peritonite/patologia , Sulfonamidas/farmacologia , Transcriptoma/genética , Animais , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Lavagem Peritoneal , Peritonite/induzido quimicamente , Peritonite/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/antagonistas & inibidores
11.
Int J Mol Sci ; 22(9)2021 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-33923285

RESUMO

Prolactin-releasing Peptide (PrRP) is a neuropeptide whose receptor is GPR10. Recently, the regulatory role of PrRP in the neuroendocrine field has attracted increasing attention. However, the influence of PrRP on macrophages, the critical housekeeper in the neuroendocrine field, has not yet been fully elucidated. Here, we investigated the effect of PrRP on the transcriptome of mouse bone marrow-derived macrophages (BMDMs) with RNA sequencing, bioinformatics, and molecular simulation. BMDMs were exposed to PrRP (18 h) and were subjected to RNA sequencing. Differentially expressed genes (DEGs) were acquired, followed by GO, KEGG, and PPI analysis. Eight qPCR-validated DEGs were chosen as hub genes. Next, the three-dimensional structures of the proteins encoded by these hub genes were modeled by Rosetta and Modeller, followed by molecular dynamics simulation by the Gromacs program. Finally, the binding modes between PrRP and hub proteins were investigated with the Rosetta program. PrRP showed no noticeable effect on the morphology of macrophages. A total of 410 DEGs were acquired, and PrRP regulated multiple BMDM-mediated functional pathways. Besides, the possible docking modes between PrRP and hub proteins were investigated. Moreover, GPR10 was expressed on the cell membrane of BMDMs, which increased after PrRP exposure. Collectively, PrRP significantly changed the transcriptome profile of BMDMs, implying that PrRP may be involved in various physiological activities mastered by macrophages.


Assuntos
Regulação da Expressão Gênica/efeitos dos fármacos , Macrófagos/metabolismo , Hormônio Liberador de Prolactina/farmacologia , Transcriptoma/efeitos dos fármacos , Animais , Biologia Computacional , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL
12.
Int J Mol Sci ; 22(5)2021 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-33652750

RESUMO

In the process of exploring space, the astronaut's body undergoes a series of physiological changes. At the level of cellular behavior, microgravity causes significant alterations, including bone loss, muscle atrophy, and cardiovascular deconditioning. At the level of gene expression, microgravity changes the expression of cytokines in many physiological processes, such as cell immunity, proliferation, and differentiation. At the level of signaling pathways, the mitogen-activated protein kinase (MAPK) signaling pathway participates in microgravity-induced immune malfunction. However, the mechanisms of these changes have not been fully elucidated. Recent studies suggest that the malfunction of macrophages is an important breakthrough for immune disorders in microgravity. As the first line of immune defense, macrophages play an essential role in maintaining homeostasis. They activate specific immune responses and participate in large numbers of physiological activities by presenting antigen and secreting cytokines. The purpose of this review is to summarize recent advances on the dysfunction of macrophages arisen from microgravity and to discuss the mechanisms of these abnormal responses. Hopefully, our work will contribute not only to the future exploration on the immune system in space, but also to the development of preventive and therapeutic drugs against the physiological consequences of spaceflight.


Assuntos
Doenças do Sistema Imunitário/imunologia , Sistema de Sinalização das MAP Quinases/imunologia , Macrófagos/imunologia , Simulação de Ausência de Peso , Ausência de Peso/efeitos adversos , Animais , Apresentação de Antígeno , Citocinas/imunologia , Humanos , Doenças do Sistema Imunitário/patologia , Macrófagos/patologia
13.
Chemistry ; 26(46): 10564-10574, 2020 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-32428289

RESUMO

Osteoarthritis is a typical degenerative joint disease related to a lubrication deficiency of articular cartilage, which is characterized by increased friction at the joint surface and severe inflammation of the joint capsule. Consequently, therapies combining lubrication restoration and drug intervention are regarded as a promising strategy for the treatment of osteoarthritis. In the present study, thermo-sensitive dual-functional nanospheres, poly[N-isopropylacrylamide-2-methacryloyloxyethyl phosphorylcholine] (PNIPAM-PMPC), are developed through emulsion polymerization. The PNIPAM-PMPC nanospheres could enhance lubrication based on the hydration lubrication mechanism by forming a tenacious hydration layer surrounding the zwitterionic headgroups, and achieve local drug delivery by encapsulating the anti-inflammatory drug diclofenac sodium. The lubrication and drug release tests showed improved lubrication and thermo-sensitive drug release of the nanospheres. The in vitro test using cytokines-treated chondrocytes indicated that the PNIPAM-PMPC nanospheres were biocompatible and upregulated anabolic genes and simultaneously downregulated catabolic genes of the articular cartilage. In summary, the developed PNIPAM-PMPC nanospheres, with the property of enhanced lubrication and local drug delivery, can be an effective nanomedicine for the treatment of osteoarthritis.


Assuntos
Nanosferas , Osteoartrite , Preparações Farmacêuticas , Acrilamidas/química , Fricção , Humanos , Lubrificantes/química , Lubrificação , Osteoartrite/tratamento farmacológico , Fosforilcolina/química
14.
Int J Mol Sci ; 21(22)2020 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-33198196

RESUMO

Osteoarthritis (OA) is one of the most prevalent chronic joint diseases worldwide, which causes a series of problems, such as joint pain, muscle atrophy, and joint deformities. Benefiting from some advances in the clinical treatment of OA, the quality of life of OA patients has been improved. However, the clinical need for more effective treatments for OA is still very urgent. Increasing findings show that macrophages are a critical breakthrough in OA therapy. Stimulated by different factors, macrophages are differentiated into two phenotypes: the pro-inflammatory M1 type and anti-inflammatory M2 type. In this study, various therapeutic reagents for macrophage-dependent OA treatment are summarized, including physical stimuli, chemical compounds, and biological molecules. Subsequently, the mechanisms of action of various approaches to modulating macrophages are discussed, and the signaling pathways underlying these treatments are interpreted. The NF-κB signaling pathway plays a vital role in the occurrence and development of macrophage-mediated OA, as NF-κB signaling pathway agonists promote the occurrence of OA, whereas NF-κB inhibitors ameliorate OA. Besides, several signaling pathways are also involved in the process of OA, including the JNK, Akt, MAPK, STAT6, Wnt/ß-catenin, and mTOR pathways. In summary, macrophage polarization is a critical node in regulating the inflammatory response of OA. Reagents targeting the polarization of macrophages can effectively inhibit inflammation in the joints, which finally relieves OA symptoms. Our work lays the foundation for the development of macrophage-targeted therapeutic molecules and helps to elucidate the role of macrophages in OA.


Assuntos
Polaridade Celular/fisiologia , Macrófagos/metabolismo , Macrófagos/fisiologia , Osteoartrite/metabolismo , Osteoartrite/fisiopatologia , Transdução de Sinais/fisiologia , Animais , Humanos , Inflamação/metabolismo , Inflamação/fisiopatologia
15.
Int J Mol Sci ; 21(12)2020 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-32599940

RESUMO

At present, studies on macrophage proteins mainly focus on biological stimuli, with less attention paid to the responses of macrophage proteins to physical stimuli, such as electric fields. Here, we exploited the electric field-sensitive hub proteins of macrophages. RAW 264.7 macrophages were treated with a direct current electric field (dcEF) (200 mV/mm) for four hours, followed by RNA-Seq analysis. Differentially expressed genes (DEGs) were obtained, followed by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) and protein-protein interaction (PPI) analysis. Eight qPCR-verified DEGs were selected. Subsequently, three-dimensional protein models of DEGs were modeled by Modeller and Rosetta, followed by molecular dynamics simulation for 200 ns with GROMACS. Finally, dcEFs (10, 50, and 500 mV/mm) were used to simulate the molecular dynamics of DEG proteins for 200 ns, followed by trajectory analysis. The dcEF has no obvious effect on RAW 264.7 morphology. A total of 689 DEGs were obtained, and enrichment analysis showed that the steroid biosynthesis pathway was most affected by the dcEF. Moreover, the three-dimensional protein structures of hub proteins were constructed, and trajectory analysis suggested that the dcEF caused an increase in the atomic motion of the protein in a dcEF-intensity-dependent manner. Overall, we provide new clues and a basis for investigating the hub proteins of macrophages in response to electric field stimulation.


Assuntos
Biomarcadores/química , Biomarcadores/metabolismo , Biologia Computacional/métodos , Eletricidade , Regulação da Expressão Gênica/efeitos da radiação , Redes Reguladoras de Genes , Macrófagos/metabolismo , Animais , Células Cultivadas , Perfilação da Expressão Gênica , Camundongos , Simulação de Dinâmica Molecular , Conformação Proteica , Mapas de Interação de Proteínas , Transcriptoma
16.
Biochemistry ; 58(6): 590-607, 2019 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-30489059

RESUMO

Intraneuronal aggregation of TDP-43 is seen in 97% of all amyotrophic lateral sclerosis cases and occurs by a poorly understood mechanism. We developed a simple in vitro model system for the study of full-length TDP-43 aggregation in solution and in protein droplets. We found that soluble, YFP-tagged full-length TDP-43 (yTDP-43) dimers can be produced by refolding in low-salt HEPES buffer; these solutions are stable for several weeks. We found that physiological electrolytes induced reversible aggregation of yTDP-43 into 10-50 nm tufted particles, without amyloid characteristics. The order of aggregation induction potency was K+ < Na+ < Mg2+ < Ca2+, which is the reverse of the Hofmeister series. The kinetics of aggregation were fit to a single-step model, and the apparent rate of aggregation was affected by yTDP-43 and NaCl concentrations. While yTDP-43 alone did not form stable liquid droplets, it partitioned into preformed Ddx4N1 droplets, showing dynamic diffusion behavior consistent with liquid-liquid phase transition, but then aggregated over time. Aggregation of yTDP-43 in droplets also occurred rapidly in response to changes in electrolyte concentrations, mirroring solution behavior. This was accompanied by changes to droplet localization and solvent exchange. Exposure to extracellular-like electrolyte conditions caused rapid aggregation at the droplet periphery. The aggregation behavior of yTDP-43 is controlled by ion-specific effects that occur at physiological concentrations, suggesting a mechanistic role for local electrolyte concentrations in TDP-43 proteinopathies.


Assuntos
Amiloide/química , Proteínas de Ligação a DNA/química , Eletrólitos/farmacologia , Gotículas Lipídicas/efeitos dos fármacos , Agregados Proteicos/efeitos dos fármacos , Amiloide/efeitos dos fármacos , Proteínas de Bactérias/metabolismo , Proteínas de Ligação a DNA/efeitos dos fármacos , Proteínas de Ligação a DNA/metabolismo , Humanos , Proteínas Luminescentes/metabolismo
17.
J Am Chem Soc ; 141(14): 5640-5644, 2019 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-30912936

RESUMO

Sodium superionic conductors are keys to develop high safety and low cost all-solid-state sodium batteries. Among developed sodium ionic conductors, antiperovskite-type ionic conductors have attracted vast interest due to their high structural tolerance and good formability. Herein, we successfully synthesize Na3OBH4 with cubic antiperovskite structure by solid-state reaction from Na2O and NaBH4. Na3OBH4 exhibits ionic conductivity of 4.4 × 10-3 S cm-1 at room temperature (1.1 × 10-2 S cm-1 at 328 K) and activation energy of 0.25 eV. The ionic conductivity is 4 orders of magnitude higher than the existing antiperovskite Na3OX (X = Cl, Br, I). It is shown that such enhancement is not only due to the specific cubic antiperovskite structure of Na3OBH4 but also because of the rotation of BH4 cluster anion. This work deepens the understanding of the antiperovskite structure and the role of cluster anions for superionic conduction.

18.
Mol Reprod Dev ; 86(2): 132-144, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30303593

RESUMO

In the present study, the next-generation sequencing technology was used to develop a transcriptome database of gonad and liver from 3-year-old male and female Amur sturgeons (Acipenser schrenckii). A total of 139,406 unigenes were generated after the Illumina Hiseq. 2500 sequence and assembled by Trinity. The differential expression analysis between male and female obtained 5,199 differentially expressed genes (DEGs) in gonad and 457 DEGs in liver. Gene Ontology enrich analysis showed that the specific DEGs of gonad play a dominant role in reproductive processes. Although the specific DEGs of liver indicated their primary responsibility for energy metabolism, the DEGs of liver and gonad co-own enriched in terms associated with reproduction suggested that liver also plays a role in sex-related differences in Amur sturgeon. Furthermore, genes related to sex-related differences were selected to validate among the four different tissues by real-time quantitative polymerase chain reaction (qRT-PCR). In addition, by trans-acting analysis, a total of 5,206 putative long noncoding RNAs (lncRNAs) and 3,490 target genes of lncRNAs were predicted from gonad and liver. Moreover, several lncRNAs targeting Mea1, Piwil1, Tdrd1, Nanos2, Ankrd49, and ZP3 may have potential regulatory effect related to gametogenesis and gonadal differentiation were identified and validated by qRT-PCR. These results suggested for the first time that lncRNAs might be one of the effect factors in regulating the differential expression of messenger RNAs associated with sex-related differences in Amur sturgeon.


Assuntos
Proteínas de Peixes/biossíntese , Peixes/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/fisiologia , RNA Longo não Codificante/biossíntese , Caracteres Sexuais , Animais , Feminino , Proteínas de Peixes/genética , Peixes/genética , Masculino , RNA Longo não Codificante/genética
19.
Langmuir ; 35(20): 6735-6741, 2019 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-31030507

RESUMO

Titanium and its alloys have long been used as implantable biomaterials in orthopedics; however, to the best of our knowledge, few studies were reported to investigate surface functionalization of titanium for enhanced lubrication and sustained drug release. In the present study, titania nanotube arrays (TNTs) were prepared by anodization as effective drug nanocarriers, using titanium as the substrate. Meanwhile, motivated by articular cartilage-inspired superlubricity and mussel-inspired adhesion, a copolymer containing both dopamine methacrylamide and 2-methacryloyloxyethyl phosphorylcholine was synthesized (DMA-MPC) and spontaneously grafted onto the TNT surface, which was validated by characterization techniques such as scanning electron microscopy, water contact angle measurements, and X-ray photoelectron spectroscopy. Additionally, the lubrication test showed that copolymer-grafted TNTs have remarkably reduced friction coefficients compared with bare TNTs. Furthermore, the drug release test demonstrated that copolymer-grafted TNTs inhibited burst drug release and achieved sustained drug release in comparison with bare TNTs. In conclusion, the bioinspired surface functionalization strategy developed here, namely DMA-MPC copolymer-grafted TNTs, can be applied to modify orthopedic biomaterials (such as titanium) for enhanced lubrication and sustained drug release.


Assuntos
Materiais Revestidos Biocompatíveis/química , Portadores de Fármacos/química , Implantes Experimentais , Nanoestruturas/química , Titânio/química , Preparações de Ação Retardada/química
20.
Langmuir ; 35(40): 13189-13195, 2019 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-31547644

RESUMO

In clinics it is extremely important for implanted devices to achieve the property of enhanced lubrication and bacterial resistance; however, such a strategy has rarely been reported in previous literature. In the present study, a surface functionalization method, motivated by articular cartilage-inspired superlubrication and mussel-inspired adhesion, was proposed to modify titanium alloy (Ti6Al4V) using the copolymer (DMA-MPC) synthesized via free radical copolymerization. The copolymer-coated Ti6Al4V (Ti6Al4V@DMA-MPC) was evaluated by X-ray photoelectron spectroscopy, water contact angle, and Raman spectra to confirm that the DMA-MPC copolymer was successfully coated onto the Ti6Al4V substrate. In addition, the tribological test, with the polystyrene microsphere and Ti6Al4V or Ti6Al4V@DMA-MPC as the tribopair, indicated that the friction coefficient was greatly reduced for Ti6Al4V@DMA-MPC. Furthermore, the bacterial resistance test showed that bacterial attachment was significantly inhibited for Ti6Al4V@DMA-MPC for the three types of bacteria tested. The enhanced lubrication and bacterial resistance of Ti6Al4V@DMA-MPC was due to the tenacious hydration shell formed surrounding the zwitterionic charges in the phosphorylcholine group of the DMA-MPC copolymer. In summary, a bioinspired surface functionalization strategy is developed in this study, which can act as a universal and promising method to achieve enhanced lubrication and bacterial resistance for biomedical implants.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Materiais Revestidos Biocompatíveis/química , Lubrificantes/farmacologia , Metacrilatos/farmacologia , Fosforilcolina/análogos & derivados , Titânio/química , Ligas , Animais , Antibacterianos/química , Antibacterianos/toxicidade , Linhagem Celular , Materiais Revestidos Biocompatíveis/toxicidade , Escherichia coli/efeitos dos fármacos , Escherichia coli/fisiologia , Lubrificantes/química , Lubrificantes/toxicidade , Lubrificação , Metacrilatos/química , Metacrilatos/toxicidade , Camundongos , Fosforilcolina/química , Fosforilcolina/farmacologia , Fosforilcolina/toxicidade , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/fisiologia , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/fisiologia , Titânio/toxicidade
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