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1.
Biomed Chromatogr ; 38(5): e5840, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38402901

RESUMO

The incidence of colibacillosis in poultry is on the rise, significantly affecting the chicken industry. Ceftiofur sodium (CS) is frequently employed to treat this disease, resulting in lipopolysaccharide (LPS) buildup. Processing plays a vital role in traditional Chinese veterinary medicine. The potential intervention in liver injury by polysaccharides from the differently processed products of Angelica sinensis (PDPPAS) induced by combined CS and LPS remains unclear. This study aims to investigate the protective effect of PDPPAS on chicken liver injury caused by CS combined with LPS buildup and further identify the polysaccharides with the highest hepatoprotective activity in chickens. Furthermore, the study elucidates polysaccharides' intervention mechanism using tandem mass tag (TMT) proteomics and multiple reaction monitoring (MRM) methods. A total of 190 1-day-old layer chickens were randomly assigned into 12 groups, of which 14 chickens were in the control group and 16 in other groups, for a 10-day trial. The screening results showed that charred A. sinensis polysaccharide (CASP) had the most effective and the best hepatoprotective effect at 48 h. TMT proteomics and MRM validation results demonstrated that the intervention mechanism of the CASP high-dose (CASPH) intervention group was closely related to the protein expressions of FCER2, TBXAS1, CD34, AGXT, GCAT, COX7A2L, and CYP2AC1. Conclusively, the intervention mechanism of CASPH had multitarget, multicenter regulatory features.


Assuntos
Angelica sinensis , Galinhas , Fígado , Polissacarídeos , Proteômica , Espectrometria de Massas em Tandem , Animais , Angelica sinensis/química , Proteômica/métodos , Polissacarídeos/farmacologia , Polissacarídeos/química , Polissacarídeos/análise , Espectrometria de Massas em Tandem/métodos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Proteoma/análise , Proteoma/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/química , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle
2.
EMBO J ; 38(18): e100948, 2019 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-31418899

RESUMO

As a ubiquitous bacterial secondary messenger, c-di-GMP plays key regulatory roles in processes such as bacterial motility and transcription regulation. CobB is the Sir2 family protein deacetylase that controls energy metabolism, chemotaxis, and DNA supercoiling in many bacteria. Using an Escherichia coli proteome microarray, we found that c-di-GMP strongly binds to CobB. Further, protein deacetylation assays showed that c-di-GMP inhibits the activity of CobB and thereby modulates the biogenesis of acetyl-CoA. Interestingly, we also found that one of the key enzymes directly involved in c-di-GMP production, DgcZ, is a substrate of CobB. Deacetylation of DgcZ by CobB enhances its activity and thus the production of c-di-GMP. Our work establishes a novel negative feedback loop linking c-di-GMP biogenesis and CobB-mediated protein deacetylation.


Assuntos
GMP Cíclico/análogos & derivados , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Fósforo-Oxigênio Liases/metabolismo , Sirtuínas/metabolismo , Acetilcoenzima A/metabolismo , Acetilação , GMP Cíclico/metabolismo , Retroalimentação Fisiológica , Regulação Bacteriana da Expressão Gênica , Análise Serial de Proteínas/métodos , Proteômica/métodos , Sistemas do Segundo Mensageiro
3.
Mol Cell Proteomics ; 20: 100059, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33109704

RESUMO

Antibodies play essential roles in both diagnostics and therapeutics. Epitope mapping is essential to understand how an antibody works and to protect intellectual property. Given the millions of antibodies for which epitope information is lacking, there is a need for high-throughput epitope mapping. To address this, we developed a strategy, Antibody binding epitope Mapping (AbMap), by combining a phage displayed peptide library with next-generation sequencing. Using AbMap, profiles of the peptides bound by 202 antibodies were determined in a single test, and linear epitopes were identified for >50% of the antibodies. Using spike protein (S1 and S2)-enriched antibodies from the convalescent serum of one COVID-19 patient as the input, both linear and potentially conformational epitopes of spike protein specific antibodies were identified. We defined peptide-binding profile of an antibody as the binding capacity (BiC). Conceptually, the BiC could serve as a systematic and functional descriptor of any antibody. Requiring at least one order of magnitude less time and money to map linear epitopes than traditional technologies, AbMap allows for high-throughput epitope mapping and creates many possibilities.


Assuntos
COVID-19/imunologia , Mapeamento de Epitopos/métodos , Glicoproteína da Espícula de Coronavírus/imunologia , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Anticorpos Antivirais/metabolismo , Ensaio de Imunoadsorção Enzimática , Epitopos/metabolismo , Proteínas de Escherichia coli/imunologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Soros Imunes/sangue , Soros Imunes/imunologia , Biblioteca de Peptídeos
4.
Arch Microbiol ; 204(3): 189, 2022 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-35194691

RESUMO

Truffles are the fruiting bodies of hypogeous fungi in the genus Tuber. Some truffle species usually grow in an area devoid of vegetation, called brûlé, but limited knowledge is available on the microbial composition and structure of them. Here, we investigated the bacterial and fungal communities of Tuber indicum ascocarps and soils inside and outside a characteristic brûlé from a poplar plantation with no truffle production history in northeastern China using a high-throughput sequencing approach. A predominance of members of the bacterial phylum Proteobacteria was observed in all samples. Members of Bacillus were the main genera in the ascocarps, while members of Lysobacter and unidentified Acidobacteria were more abundant in the soil. In addition, members of Gibberella, Fusarium, and Absidia were the dominant fungi in the ascocarps, while members of Tuber were enriched in the ascocarps and soils inside the brûlé. Some mycorrhization helper bacteria (Rhizobium) and ectomycorrhiza-associated bacteria (Lysobacter) were detected, indicating their potential roles in the complex development of underground fruiting bodies and brûlé formation. These findings may contribute to the protection and cultivation of truffles.


Assuntos
Ascomicetos , Microbiota , Ascomicetos/genética , Solo , Microbiologia do Solo
5.
Plant Cell Rep ; 41(10): 2037-2088, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35904590

RESUMO

KEY MESSAGE: Forty PaCRKs have been identified from sweet cherry and overexpression PaCRK1 in sweet cherry enhances its resistance to salt stress. Cysteine-rich receptor-like kinases (CRKs), a large subgroup of the receptor-like kinases, play an important role in plant development and stress response. However, knowledge about CRKs and its function against adverse environmental stresses in sweet cherry were lacking. In this study, 40 PaCRKs were identified from sweet cherry (Prunus avium) genome database. Phylogenetic analysis indicated that PaCRKs could be classified into six subgroups. Transcriptome analysis showed that the expression levels of most PaCRKs were changed under external environmental stresses. Functional study showed that PaCRK1 overexpression could enhance Arabidopsis and sweet cherry tolerance to salt stress. Moreover, biochemical analysis showed that PaCRK1 increased salt tolerance of sweet cherry by regulating the expression of antioxidation-related genes and their enzyme activities. This study provides a comprehensive understanding of PaCRKs in sweet cherry and elucidates the potential role of PaCRKs in response to various environmental stimuli.


Assuntos
Arabidopsis , Prunus avium , Arabidopsis/genética , Cisteína/metabolismo , Filogenia , Prunus avium/genética , Tolerância ao Sal/genética
6.
Biomed Chromatogr ; 36(6): e5362, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35393691

RESUMO

Chicken colibacillosis is one of the most severe diseases in the poultry industry. Ceftiofur sodium (CS) is often used to treat it in clinical practice and lipopolysaccharide (LPS) accumulates in the chicken's body. Previous experimental studies found that CS combined with LPS could induce liver injury in layer chickens, and polysaccharides from charred Angelica sinensis(CASP) had a better hepatoprotective effect than polysaccharides from unprocessed Angelica sinensis(UASP). However, the intervention mechanism was unclear. Thus, UPLC-Q/TOF-MS/MS-based metabonomics and transcriptomics were used in this study to clarify the hepatoprotective effect mechanism of CASP and UASP in layer chickens. Transcriptomics and enzyme-linked immunosorbent assay were used for biological verification of some critical mutual metabolic pathways screened with metabonomics. The comprehensive analysis results showed that in a layer chicken liver injury model built with LPS and CS, 12 critical metabolic pathways were disturbed, involving 10 important differential metabolites. The hepatoprotective effect mechanism of CASP is related to the arachidonic acid metabolism and mTOR signaling pathways, involving nine important differential metabolites. In contrast, the hepatoprotective effect mechanism of UASP is related to the arachidonic acid metabolism pathway, involving six important differential metabolites.


Assuntos
Angelica sinensis , Animais , Ácido Araquidônico , Galinhas , Lipopolissacarídeos , Metabolômica/métodos , Polissacarídeos/farmacologia , Espectrometria de Massas em Tandem
7.
Biomed Chromatogr ; 36(2): e5252, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34591996

RESUMO

Angelica sinensis (AS) is a common Traditional Chinese Medicine used for tonifying blood in China. Unprocessed AS and its four kinds of processed products (ASs) are used to treat blood deficiency syndrome in the country. The different blood-tonifying mechanisms of ASs remain unclear. In this work, a novel method integrating metabolomics and hematological and biochemical parameters was established to provide a complementary explanation of blood supplementation mechanism of ASs. Our results revealed that different ASs exhibited various blood supplementation effect, and that AS parched with alcohol demonstrated the best blood supplementation effect. Eight metabolites from liver tissue and 12 metabolites from spleen tissue were considered to be potential biomarkers. These biomarkers were involved in four metabolic pathways. Correlation analysis results showed that l-aspartic acid and l-alanine (spleen tissue), linoleic acid, and l-cystathionine (liver tissue) exhibited a high positive or negative correlation with the aforesaid biochemical indicators. The blood-supplementation effect mechanism of ASs were related to four metabolic pathways. l-Aspartic acid and l-alanine (spleen tissue), linoleic acid, and l-cystathionine (liver tissue) were the four key metabolites associated with the blood supplementation effect of ASs. This study gives a complementary explanation of the blood supplementation effect and mechanism of action of ASs.


Assuntos
Angelica sinensis/química , Medicamentos de Ervas Chinesas/farmacologia , Medicina Tradicional Chinesa , Metaboloma/efeitos dos fármacos , Aminoácidos/metabolismo , Animais , Cromatografia Gasosa-Espectrometria de Massas , Ácido Linoleico/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Metabolômica/métodos , Camundongos , Baço/efeitos dos fármacos , Baço/metabolismo
8.
Mol Cell Proteomics ; 18(9): 1851-1863, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31308251

RESUMO

Systemic lupus erythematosus (SLE) is one of the most serious autoimmune diseases, characterized by highly diverse clinical manifestations. A biomarker is still needed for accurate diagnostics. SLE serum autoantibodies were discovered and validated using serum samples from independent sample cohorts encompassing 306 participants divided into three groups, i.e. healthy, SLE patients, and other autoimmune-related diseases. To discover biomarkers for SLE, a phage displayed random peptide library (Ph.D. 12) and deep sequencing were applied to screen specific autoantibodies in a total of 100 serum samples from 50 SLE patients and 50 healthy controls. A statistical analysis protocol was set up for the identification of peptides as potential biomarkers. For validation, 10 peptides were analyzed using enzyme-linked immunosorbent assays (ELISA). As a result, four peptides (SLE2018Val001, SLE2018Val002, SLE2018Val006, and SLE2018Val008) were discovered with high diagnostic power to differentiate SLE patients from healthy controls. Among them, two peptides, i.e. SLE2018Val001 and SLE2018Val002, were confirmed between SLE with other autoimmune patients. The procedure we established could be easily adopted for the identification of autoantibodies as biomarkers for many other diseases.


Assuntos
Lúpus Eritematoso Sistêmico/sangue , Biblioteca de Peptídeos , Peptídeos/sangue , Adulto , Área Sob a Curva , Doenças Autoimunes/sangue , Biomarcadores/sangue , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Peptídeos/genética , Reprodutibilidade dos Testes
9.
Acta Biochim Biophys Sin (Shanghai) ; 53(5): 628-635, 2021 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-33637989

RESUMO

PD-1 plays an important role as an immune checkpoint. Sintilimab is a newly approved PD-1 antibody for cancer immunotherapy with an unknown binding epitope on PD-1. In this study, to elucidate the molecular mechanism by which sintilimab blocks PD-1 activation, we applied Antibody binding epitope Mapping (AbMap) to identify the binding epitope of sintilimab. An epitope was successfully identified, i.e. SLAPKA, aa 127-132. By constructing a series of point mutations, the dominant residues S127, L128, A129, P130, and A132 of PD-1 were further validated by western blot analysis, biolayer interferometry, and flow cytometry. Structural analysis showed that the epitope is partially within the binding interface of PD-1 and PD-L1, and this epitope also partially overlaps with that of nivolumab and pembrolizumab. These results demonstrate that sintilimab can attenuate PD-1 activation by directly competing with the interaction between PD-1 and PD-L1 through binding with the key residues of the FG loop on PD-1. This study also demonstrates the high efficiency and accuracy of AbMap for determining the binding epitope of therapeutic antibodies.


Assuntos
Anticorpos Monoclonais Humanizados/química , Antineoplásicos Imunológicos/química , Mapeamento de Epitopos , Epitopos/química , Receptor de Morte Celular Programada 1/química , Anticorpos Monoclonais Humanizados/imunologia , Antineoplásicos Imunológicos/imunologia , Epitopos/imunologia , Humanos , Receptor de Morte Celular Programada 1/imunologia
10.
Pharm Biol ; 58(1): 1167-1176, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33222580

RESUMO

CONTEXT: Angelica sinensis (Oliv.) Diels (Apiaceae) (syn. Angelica polymorpha Maxim var. sinensis Oliver) processed with yellow rice wine (WAS) has a blood-supplementing effect. OBJECTIVE: To establish an optimal technology for preparing water decoction of WAS (WASD), and screen blood-supplementing fractions. MATERIALS AND METHODS: Ferulic acid and crude polysaccharide were used in optimizing the preparation technology for WASD through response surface methodology. The independent variables were liquid-solid ratio, soaking time, and extraction time. Eighty Kunming mice were randomly divided into normal control, model, and six intervention groups (n = 10). The intervention groups were given different WASD fractions by gavage (5 or 10 g/kg). The model intervention groups received acetylphenyl hydrazine (subcutaneous injection) and cyclophosphamide (intraperitoneal injection). Duration of study, 9 days. The components of blood-supplementing fractions were analyzed. RESULTS: The optimum extraction parameters were liquid-solid ratio, 7.69:1 mL/g; soaking time, 119.78 min; and extraction time, 143.35 min. The optimal OD value was 0.8437. RBC, WBC, and Hb in the water fraction (5, 10 g/kg) and n-butanol fraction (10 g/kg) intervention groups increased significantly compared with the model group (p < 0.05). Polysaccharide and caffeic acid contents of water fraction were 252.565 and 0.346 µg/mg, respectively; ferulic acid was not detected. Caffeic acid and ferulic acid contents of n-butanol fraction were 1.187 and 0.806 µg/mg, respectively, polysaccharide was not detected. CONCLUSIONS: The optimum preparation technology of WASD was obtained, and the water, n-butanol fractions were blood-supplementing fractions. This study provides a theoretical foundation for further application of WAS in the pharmaceutical industry.


Assuntos
Angelica sinensis/química , Sangue/efeitos dos fármacos , Oryza/química , Extratos Vegetais/farmacologia , Animais , Contagem de Células Sanguíneas , Ácidos Cafeicos/química , Ácidos Cafeicos/farmacologia , Cromatografia Líquida de Alta Pressão , Ácidos Cumáricos/química , Ácidos Cumáricos/farmacologia , Medicina Tradicional Chinesa , Camundongos , Raízes de Plantas/química , Polissacarídeos/química , Polissacarídeos/farmacologia , Solventes , Espectrofotometria Ultravioleta , Timo/efeitos dos fármacos , Água , Vinho
11.
Mol Cell Proteomics ; 16(8): 1491-1506, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28572091

RESUMO

Mycobacterium tuberculosis (Mtb) is the causative agent of tuberculosis, the leading cause of death among all infectious diseases. There are 11 eukaryotic-like serine/threonine protein kinases (STPKs) in Mtb, which are thought to play pivotal roles in cell growth, signal transduction and pathogenesis. However, their underlying mechanisms of action remain largely uncharacterized. In this study, using a Mtb proteome microarray, we have globally identified the binding proteins in Mtb for all of the STPKs, and constructed the first STPK protein interaction (KPI) map that includes 492 binding proteins and 1,027 interactions. Bioinformatics analysis showed that the interacting proteins reflect diverse functions, including roles in two-component system, transcription, protein degradation, and cell wall integrity. Functional investigations confirmed that PknG regulates cell wall integrity through key components of peptidoglycan (PG) biosynthesis, e.g. MurC. The global STPK-KPIs network constructed here is expected to serve as a rich resource for understanding the key signaling pathways in Mtb, thus facilitating drug development and effective control of Mtb.


Assuntos
Proteínas de Bactérias/metabolismo , Mycobacterium tuberculosis/metabolismo , Mapas de Interação de Proteínas , Proteínas Serina-Treonina Quinases/metabolismo , Proteoma/metabolismo , Proteínas de Bactérias/genética , Parede Celular , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/patogenicidade , Fosforilação , Proteínas Serina-Treonina Quinases/genética , Proteoma/genética , Proteômica , Transdução de Sinais
12.
Mol Cell Proteomics ; 16(12): 2243-2253, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29018126

RESUMO

Mycobacterium tuberculosis (Mtb) has evolved multiple strategies to counter the human immune system. The effectors of Mtb play important roles in the interactions with the host. However, because of the lack of highly efficient strategies, there are only a handful of known Mtb effectors, thus hampering our understanding of Mtb pathogenesis. In this study, we probed Mtb proteome microarray with biotinylated whole-cell lysates of human macrophages, identifying 26 Mtb membrane proteins and secreted proteins that bind to macrophage proteins. Combining GST pull-down with mass spectroscopy then enabled the specific identification of all binders. We refer to this proteome microarray-based strategy as SOPHIE (Systematic unlOcking of Pathogen and Host Interacting Effectors). Detailed investigation of a novel effector identified here, the iron storage protein BfrB (Rv3841), revealed that BfrB inhibits NF-κB-dependent transcription through binding and reducing the nuclear abundance of the ribosomal protein S3 (RPS3), which is a functional subunit of NF- κB. The importance of this interaction was evidenced by the promotion of survival in macrophages of the mycobacteria, Mycobacterium smegmatis, by overexpression of BfrB. Thus, beyond demonstrating the power of SOPHIE in the discovery of novel effectors of human pathogens, we expect that the set of Mtb effectors identified in this work will greatly facilitate the understanding of the pathogenesis of Mtb, possibly leading to additional potential molecular targets in the battle against tuberculosis.


Assuntos
Proteínas de Bactérias/metabolismo , Grupo dos Citocromos b/metabolismo , Ferritinas/metabolismo , Macrófagos/microbiologia , Mycobacterium tuberculosis/patogenicidade , Proteômica/métodos , Proteínas Ribossômicas/metabolismo , Proteínas de Bactérias/química , Sítios de Ligação , Linhagem Celular , Cristalografia por Raios X , Grupo dos Citocromos b/química , Ferritinas/química , Células HEK293 , Humanos , Imunidade Inata , Macrófagos/citologia , Macrófagos/metabolismo , Espectrometria de Massas , Modelos Moleculares , Mycobacterium tuberculosis/metabolismo , NF-kappa B/metabolismo , Análise Serial de Proteínas/métodos , Ligação Proteica , Proteínas Ribossômicas/química , Células THP-1
14.
Proteomics ; 18(23): e1800265, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30281201

RESUMO

Mycobacterium tuberculosis (Mtb) serine/threonine kinase PknG plays an important role in the Mtb-host interaction by facilitating the survival of Mtb in macrophages. However, the human proteins with which the PknG interacts, and the underlying molecular mechanisms are still largely unknown. In this study, a HuProt array is been applied to globally identify the host proteins to which PknG binds. In this way, 125 interactors are discovered, including a cyclophilin protein, CypA. This interaction between PknG and CypA is validated both in vitro and in vivo, and functional studies show that PknG significantly reduces the protein levels of CypA through phosphorylation, which consequently inhibit the inflammatory response through downregulation of NF-κB and ERK1/2 pathways. Phenotypically, overexpression of PknG reduces cytokine levels and promotes the survival of Mycobacterium smegmatis (Msm) in macrophages. Overall, it is expected that the PknG interactors identified in this study will serve as a useful resource for further systematic studies of the roles that PknG plays in the Mtb-host interactions.


Assuntos
Mycobacterium tuberculosis/metabolismo , Proteoma/análise , Proteínas de Bactérias/metabolismo , Humanos , Sistema de Sinalização das MAP Quinases , Macrófagos/metabolismo , NF-kappa B/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo
15.
Acta Biochim Biophys Sin (Shanghai) ; 46(7): 548-55, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24907045

RESUMO

Protein acetylation is one of the most abundant post-translational modifications and plays critical roles in many important biological processes. Based on the recent advances in mass spectrometry technology, in bacteria, such as Escherichia coli, tremendous acetylated proteins and acetylation sites have been identified. However, only one protein deacetylase, i.e. CobB, has been identified in E. coli so far. How CobB is regulated is still elusive. One right strategy to study the regulation of CobB is to globally identify its interacting proteins. In this study, we used a proteome microarray containing ∼4000 affinity-purified E. coli proteins to globally identify CobB interactors, and finally identified 183 binding proteins of high stringency. Bioinformatics analysis showed that these interacting proteins play a variety of roles in a wide range of cellular functions and are highly enriched in carboxylic acid metabolic process and hexose catabolic process, and also enriched in transferase and hydrolase. We further used bio-layer interferometry to analyze the interaction and quantify the kinetic parameters of putative CobB interactors, and clearly showed that CobB could strongly interact with TopA and AccC. The novel CobB interactors that we identified could serve as a start point for further functional analysis.


Assuntos
Proteínas de Escherichia coli/metabolismo , Análise Serial de Proteínas , Proteoma , Ligação Proteica
16.
Front Cell Infect Microbiol ; 14: 1427763, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39006744

RESUMO

Introduction: Rumen acidosis is one of the most common diseases in beef cattle. It severely affects the normal development of calves and poses a significant threat to the farming industry. However, the influence of rumen acidosis on the gut microbiota and serum metabolites of calves is currently unclear. Objective: The aim of this study is to investigate the changes in the gut microbiota and serum metabolites in calves after rumen acidosis and analyse the correlation. Methods: Eight calves were selected as the rumen acidosis group, and eight health calves were selected as the healthy group. The faecal gut microbiota and serum metabolites of calves were detected respectively using 16S rDNA high-throughput sequencing and non-target metabolomics. The correlation between gut microbiota and serum metabolites was analyzed by Spearman correlation analysis. Results: Differential analysis of the diversity and composition of gut microbiota between eight male healthy (Health) and eight male rumen acidosis (Disease) calves revealed that rumen acidosis increased the abundance of the gut microbiota in calves. At the phylum level, compared to the Healthy group, the relative abundance of Proteobacteria in the Disease group significantly decreased (P<0.05), while the relative abundance of Desulfobacterota significantly increased in the Disease group (P<0.05). At the genus level, compared to the Disease group, the relative abundance of Alloprevotella, Muribaculaceae, Succinivibrio, Prevotella, Agathobacter and Parabacteroides significantly increased in the Healthy group (P<0.05), while the relative abundance of Christensenellaceae_R-7 and Monoglobus significantly decreased in the Healthy group (P<0.05). Differential analysis results showed the Healthy group had 23 genera with higher abundance, while the Disease group had 47 genera with higher abundance. Serum metabolomics results revealed the differential metabolites associated with rumen acidosis, including nicotinamide, niacin, L-glutamic acid and carnosine, were mainly enriched in the nicotinate and nicotinamide pathway and the histidine pathway. Conclusion: The occurrence of rumen acidosis can induce changes in the gut microbiota of calves, with a significant increase of the Christensenellaceae_R-7 genus and a significant decrease of Prevotella and Succinivibrio genera. In addition, the occurrence of rumen acidosis can also induce changes in serum metabolites including niacin, niacinamide, L-glutamine, and carnosine, which may serve as the diagnostic biomarkers of rumen acidosis of calves.


Assuntos
Acidose , Doenças dos Bovinos , Fezes , Microbioma Gastrointestinal , Metabolômica , RNA Ribossômico 16S , Rúmen , Animais , Bovinos , Rúmen/microbiologia , Acidose/veterinária , Acidose/microbiologia , Acidose/sangue , RNA Ribossômico 16S/genética , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/sangue , Masculino , Fezes/microbiologia , DNA Ribossômico/genética , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , DNA Bacteriano/genética
17.
Tree Physiol ; 44(9)2024 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-39163268

RESUMO

Drought is a significant global issue affecting agricultural production, and the utilization of beneficial rhizosphere microorganisms is one of the effective ways to increase the productivity of crops and forest under drought. In this study, we characterized a novel growth-promoting dark septate endophytes (DSE) fungus R16 (Dothideomycetes sp.) derived from blueberry roots. Hyphae or microsclerotia were visible within the epidermal or cortical cells of R16-colonized blueberry roots, which was consistent with the typical characteristics of DSE fungi. Inoculation with R16 promoted the growth of blueberry seedlings, and the advantage over the control group was more significant under PEG-induced drought. Comparison of physiological indicators related to drought resistance between the inoculated and control groups was performed on the potted blueberry plants, including the chlorophyll content, net photosynthetic rate, root activities, malondialdehyde and H2O2 content, which indicated that R16 colonization mitigated drought injury in blueberry plants. We further analyzed the effects of R16 on phytohormones and non-structural carbohydrates (NSCs) to explore the mechanism of increased drought tolerance by R16 in blueberry seedlings. The results showed that except for the gibberellin content, indole-3-acetic acid, zeatin and abscisic acid varied significantly between the inoculated and control groups. Sucrose phosphate synthase and sorbitol-6-phosphate dehydrogenase activities in mature leaves, the key enzymes responsible for sucrose and sorbitol synthesis, respectively, as well as sorbitol dehydrogenase, sucrose synthase, cell wall invertase, hexokinase and fructokinase in roots, the key enzymes involved in the NSCs metabolism, showed significant differences between the inoculated and control groups before and after drought treatment. These results suggested that the positive effects of R16 colonization on the drought tolerance of blueberry seedlings are partially attributable to the regulation of phytohormone and sugar metabolism. This study provided valuable information for the research on the interaction between DSE fungi and host plants as well as the application of DSE preparations in agriculture.


Assuntos
Mirtilos Azuis (Planta) , Endófitos , Reguladores de Crescimento de Plantas , Mirtilos Azuis (Planta)/microbiologia , Mirtilos Azuis (Planta)/fisiologia , Mirtilos Azuis (Planta)/crescimento & desenvolvimento , Reguladores de Crescimento de Plantas/metabolismo , Endófitos/fisiologia , Secas , Raízes de Plantas/microbiologia , Raízes de Plantas/fisiologia , Raízes de Plantas/crescimento & desenvolvimento , Ascomicetos/fisiologia , Metabolismo dos Carboidratos , Plântula/fisiologia , Plântula/crescimento & desenvolvimento , Plântula/microbiologia , Resistência à Seca
18.
Tree Physiol ; 44(6)2024 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-38775231

RESUMO

Plant biomass is a highly promising renewable feedstock for the production of biofuels, chemicals and materials. Enhancing the content of plant biomass through endophyte symbiosis can effectively reduce economic and technological barriers in industrial production. In this study, we found that symbiosis with the dark septate endophyte (DSE) Anteaglonium sp. T010 significantly promoted the growth of poplar trees and increased plant biomass, including cellulose, lignin and starch. To further investigate whether plant biomass was related to sucrose metabolism, we analyzed the levels of relevant sugars and enzyme activities. During the symbiosis of Anteaglonium sp. T010, sucrose, fructose and glucose levels in the stem of poplar decreased, while the content of intermediates such as glucose-6-phosphate (G6P), fructose-6-phosphate (F6P) and UDP-glucose (UDPG), and the activity of enzymes related to sucrose metabolism, including sucrose synthase (SUSY), cell wall invertase (CWINV), fructokinase (FRK) and hexokinase, increased. In addition, the contents of glucose, fructose, starch, and their intermediates G6P, F6P and UDPG, as well as the enzyme activities of SUSY, CWINV, neutral invertase and FRK in roots were increased, which ultimately led to the increase of root biomass. Besides that, during the symbiotic process of Anteaglonium sp. T010, there were significant changes in the expression levels of root-related hormones, which may promote changes in sucrose metabolism and consequently increase the plant biomass. Therefore, this study suggested that DSE fungi can increase the plant biomass synthesis capacity by regulating the carbohydrate allocation and sink strength in poplar.


Assuntos
Biomassa , Endófitos , Populus , Sacarose , Populus/metabolismo , Populus/crescimento & desenvolvimento , Populus/microbiologia , Sacarose/metabolismo , Endófitos/fisiologia , Endófitos/metabolismo , Ascomicetos/fisiologia , Reguladores de Crescimento de Plantas/metabolismo , Simbiose
19.
Plant Physiol Biochem ; 216: 109125, 2024 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-39278049

RESUMO

Soil salinization is a major environmental factor that severely affects global agriculture. Root endophytes can enter root cells, and offer various ecological benefits, such as promoting plant growth, improving soil conditions, and enhancing plant resistance. Su100 is a novel strain of endophytic fungus that was characterized from blueberry roots. In this study, we focused on evaluating the effects of Su100 secretion on maize growth. The results demonstrated that maize treated with Su100 fermentation broth (SFB) exhibited significantly stronger salt tolerance than the control. It is worth mentioning that the treated root system not only had an advantage in terms of biomass but also a change in root structure with a significant increase in lateral roots (LRs) compared to the control. Transcriptome analysis combined with hormone content measurements indicated that SFB upregulated the auxin signaling pathway, and also caused alterations in brassinosteroids (BR) and jasmonic acid (JA) biosynthesis and signaling pathways. Transcriptome analyses also indicated that SFB caused significant changes in the sugar metabolism of maize roots. The major changes included: enhancing the conversion and utilization of sucrose in roots; increasing carbon flow to uridine diphosphate glucose (UDPG), which acted as a precursor for producing more cell wall polysaccharides, mainly pectin and lignin; accelerating the tricarboxylic acid cycle, which were further supported by sugar content determinations. Taken together, our results indicated that the enhanced salt tolerance of maize treated with SFB was due to the modulation of sugar metabolism and phytohormone biosynthesis or signaling pathways. This study provided new insights into the mechanisms of action of endophytic fungi and highlighted the potential application of fungal preparations in agriculture.

20.
Food Chem Toxicol ; 190: 114796, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38852756

RESUMO

Pulmonary fibrosis is the outcome of the prolonged interstitial pneumonia, characterized by excessive accumulation of fibroblasts and collagen deposition, leading to its development. This study aimed to study the changes in PI3K/AKT and NRF2/HO-1 signaling expression and intestinal microbiota in a rat model of a novel bleomycin-induced pulmonary fibrosis. The findings of our study showed the model was successfully established. The results showed that the alveolar septum in the model was significantly widened and infiltrated by severe inflammatory cells. Alveolar atrophy occurred due to the formation of multiple inflammatory foci. During this period, fibrous tissue was distributed in strips and patches, primarily around the pulmonary interstitium and bronchus. Moreover, lung damage and fibrosis progressively worsened over time. The mRNA expression of HO-1 and NRF2 in the model decreased while the mRNA expression of HIF-1α, VEGF, PI3K and AKT increased. Furthermore, it was observed to decrease the protein expression of E-cad, HO-1 and NRF2, and increase the protein expression of α-SMA and p-AKT. Additionally, this model leaded to an imbalance in the intestinal microbiota. This study demonstrate that the novel pulmonary fibrosis model activates the NRF2/HO-1 pathway and the PI3K/AKT pathway in rat lung tissues, and leading to intestinal barrier disorder.


Assuntos
Bleomicina , Microbioma Gastrointestinal , Fator 2 Relacionado a NF-E2 , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Fibrose Pulmonar , Transdução de Sinais , Animais , Masculino , Ratos , Heme Oxigenase (Desciclizante)/metabolismo , Heme Oxigenase (Desciclizante)/genética , Heme Oxigenase-1/metabolismo , Heme Oxigenase-1/genética , Fator 2 Relacionado a NF-E2/metabolismo , Fator 2 Relacionado a NF-E2/genética , Fosfatidilinositol 3-Quinases/metabolismo , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/metabolismo , Ratos Sprague-Dawley
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