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Fatty liver injury is characterized by liver fat accumulation and results in serious health problems worldwide. There is no effective treatment that reverses fatty liver injury besides etiological therapy. Inflammation is an important macrophage-involving pathological process of liver injury. Here, we investigated the role of sphingosine 1-phosphate receptors (S1PRs) in fatty liver injury and explored whether S1PR2/3 blockade could cure fatty liver injury. A methionine-choline-deficient and a high-fat (MCDHF) diet was used to induce fatty liver injury, and the number of macrophages was evaluated by flow cytometry. Gene expressions were detected using RT-qPCR and cytometric bead array. In MCDHF-diet-fed mice, pro-inflammatory factor expressions were upregulated by fatty liver injury. The S1P level and S1PR2/3 expressions were significantly elevated. Moreover, increased S1P level and S1PR2/3 mRNA expressions were positively correlated with pro-inflammatory factor expressions in the liver. Furthermore, the number of pro-inflammatory macrophages (iMφ) increased in injured liver, and they were mainly bone-marrow-derived macrophages. In vivo, S1PR2/3 blockade decreased the amount of iMφ and inflammation and attenuated liver injury and fibrosis, although liver fat accumulation was unchanged. These data strongly suggest that anti-inflammatory treatment by blocking the S1P/S1PR2/3 axis attenuates fatty liver injury, which might serve as a potential target for fatty liver injury.
Assuntos
Fígado Gorduroso/etiologia , Fígado Gorduroso/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Receptores de Esfingosina-1-Fosfato/antagonistas & inibidores , Animais , Biópsia , Células Cultivadas , Dieta Hiperlipídica , Modelos Animais de Doenças , Fígado Gorduroso/patologia , Fibrose , Inflamação/etiologia , Inflamação/metabolismo , Inflamação/patologia , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Receptores de Esfingosina-1-Fosfato/metabolismoRESUMO
(1) Background: Ionic liquids (ILs) are considered "green" solvents and have been widely used in the extraction and separation field in recent years; (2) Methods: In this study, some common ILs and functionalized magnetic ionic liquids (MILs) were used as adjuvants for the solvent extraction of paclitaxel from Taxus x media (T. x media) using methanol solution. The extraction conditions of methanol concentration, IL type and amount, solid-liquid ratio, extraction temperature, and ultrasonic irradiation time were investigated in single factor experiments. Then, three factors of IL amount, solid-liquid ratio, and ultrasonic irradiation time were optimized by response surface methodology (RSM); (3) Results: The MIL [C4MIM]FeCl3Br was screened as the optimal adjuvant. Under the optimization conditions of 1.2% IL amount, 1:10.5 solid-liquid ratio, and 30 min ultrasonic irradiation time, the extraction yield reached 0.224 mg/g; and (4) Conclusions: Compared with the conventional solvent extraction, this ultrasonic assisted extraction (UAE) using methanol and MIL as adjuvants can significantly improve the extraction yield, reduce the use of methanol, and shorten the extraction time, which has the potentiality of being used in the extraction of some other important bioactive compounds from natural plant resources.
Assuntos
Antineoplásicos Fitogênicos/isolamento & purificação , Líquidos Iônicos/química , Extração Líquido-Líquido/métodos , Paclitaxel/isolamento & purificação , Taxus/química , Análise Fatorial , Concentração de Íons de Hidrogênio , Extração Líquido-Líquido/economia , Metanol/química , Extratos Vegetais/química , Solventes/química , Sonicação , Temperatura , Fatores de TempoRESUMO
Intercropping is a widely used agricultural system; however, the effect of intercropping between accumulator plants on phytoextraction in heavy metal-contaminated soils remains unknown. Here, a field experiment was conducted to investigate the phytoextraction efficiency and related environmental effects of three Amaranthaceae plants (Amaranthus hypochondriacus, Celosia argentea, and Pfaffia glomerata) using mono- and intercropping models. In monocropping, the total biomass of A. hypochondriacus was only 51.2 % of that of C. argentea. Compared with monocropping, intercropping reduced the fresh weight per plant of A. hypochondriacus by 53.0 % (intercropping with C. argentea) and 40.5 % (intercropping with P. glomerata) but increased the biomass per plant of C. argentea and P. glomerata by 128.2 and 14.2 %, respectively. The Cd uptake of the three plants in the monocropping models showed the following trend: C. argentea > P. glomerata > A. hypochondriacus. Interplanting A. hypochondriacus and C. argentea further increased the phytoextraction efficiency by 361.2 % (compared with A. hypochondriacus monocropping) and 52.0 % (compared with C. argentea monocropping). Soil exchangeable Cd, Pb, Cu, Zn, K, and P, soil N-NO3- and N-NH4+, soil common bacteria and arbuscular mycorrhiza (AM) fungi, and soil total organic carbon (TOC) play key roles in Cd and Pb uptake by the three accumulator plants (p < 0.05). The biomass of common bacteria, Gm+, Gm- bacteria, fungi, AM fungi, and actinomycetes increased with the three accumulators planted in the mono- and intercropping models. Compared with C. argentea monocropping, the biomass of soil microbes in the rhizosphere soil was obviously increased in the intercropping A. hypochondriacus and C. argentea models. These results suggest that interplanting A. hypochondriacus and C. argentea can increase Cd removal efficiency from Cd-contaminated soils, and this model could be recommended to remediate Cd-contaminated soils on a field scale.
Assuntos
Amaranthus , Poluentes do Solo , Cádmio/análise , Biodegradação Ambiental , Chumbo , Poluentes do Solo/análise , Solo , PlantasRESUMO
The gut microbiota and diet-induced changes in microbiome composition have been linked to various liver diseases, although the specific microbes and mechanisms remain understudied. Alcohol-related liver disease (ALD) is one such disease with limited therapeutic options due to its complex pathogenesis. We demonstrate that a diet rich in soluble dietary fiber increases the abundance of Bacteroides acidifaciens (B. acidifaciens) and alleviates alcohol-induced liver injury in mice. B. acidifaciens treatment alone ameliorates liver injury through a bile salt hydrolase that generates unconjugated bile acids to activate intestinal farnesoid X receptor (FXR) and its downstream target, fibroblast growth factor-15 (FGF15). FGF15 promotes hepatocyte expression of ornithine aminotransferase (OAT), which facilitates the metabolism of accumulated ornithine in the liver into glutamate, thereby providing sufficient glutamate for ammonia detoxification via the glutamine synthesis pathway. Collectively, these findings uncover a potential therapeutic strategy for ALD involving dietary fiber supplementation and B. acidifaciens.
Assuntos
Amônia , Bacteroides , Fibras na Dieta , Fatores de Crescimento de Fibroblastos , Microbioma Gastrointestinal , Camundongos Endogâmicos C57BL , Animais , Bacteroides/metabolismo , Camundongos , Fibras na Dieta/metabolismo , Amônia/metabolismo , Microbioma Gastrointestinal/fisiologia , Fatores de Crescimento de Fibroblastos/metabolismo , Hepatopatias Alcoólicas/metabolismo , Hepatopatias Alcoólicas/microbiologia , Masculino , Fígado/metabolismo , Hepatócitos/metabolismo , Ácidos e Sais Biliares/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Humanos , Inativação Metabólica , AmidoidrolasesRESUMO
Healthy Kupffer cell (KC) pool is dominated by embryonic KCs (EmKCs), preserving liver homeostasis. How the KC pool varies upon injury remains unclear. Using chimeric mice with bone marrow (BM) cells labeled with enhanced green fluorescent protein, we identify that BM monocyte-derived KCs (MoKCs) become dominant in cholestatic- or toxic-injured livers via immunofluorescence and mass cytometry. Single-cell RNA sequencing (scRNA-seq) unveils the enhanced proliferative, anti-apoptotic properties and repair potential of MoKCs compared with EmKCs, which are confirmed in vivo and ex vivo through flow cytometry, qPCR, Cell Counting Kit-8, and immunofluorescence. Furthermore, compared with EmKC-dominated livers, MoKC-dominated livers exhibit less functional damage, necrosis, and fibrosis under damage, as tested by serum alanine aminotransferase activity detection, H&E and Sirius red staining, qPCR, and western blot. Collectively, we highlight that MoKCs dominate the KC pool in injured livers and show enhanced proliferative and anti-apoptotic properties while also promoting repair and attenuating fibrosis.
Assuntos
Células de Kupffer , Monócitos , Camundongos , Animais , Células de Kupffer/metabolismo , Fígado/metabolismo , Necrose/metabolismo , FibroseRESUMO
15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) exhibited potential to alleviate liver inflammation in chronic injury but was less studied in acute injury. Acute liver injury was associated with elevated macrophage migration inhibitory factor (MIF) levels in damaged hepatocytes. This study aimed to investigate the regulatory mechanism of hepatocyte-derived MIF by 15d-PGJ2 and its subsequent impact on acute liver injury. In vivo, mouse models were established by carbon tetrachloride (CCl4) intraperitoneal injection, with or without 15d-PGJ2 administration. 15d-PGJ2 treatment reduced the necrotic areas induced by CCl4. In the same mouse model constructed using enhanced green fluorescent protein (EGFP)-labeled bone marrow (BM) chimeric mice, 15d-PGJ2 reduced CCl4 induced BM-derived macrophage (BMM, EGFP+F4/80+) infiltration and inflammatory cytokine expression. Additionally, 15d-PGJ2 down-regulated liver and serum MIF levels; liver MIF expression was positively correlated with BMM percentage and inflammatory cytokine expression. In vitro, 15d-PGJ2 inhibited Mif expression in hepatocytes. In primary hepatocytes, reactive oxygen species inhibitor (NAC) showed no effect on MIF inhibition by 15d-PGJ2; PPARγ inhibitor (GW9662) abolished 15d-PGJ2 suppressed MIF expression and antagonists (troglitazone, ciglitazone) mimicked its function. In Pparg silenced AML12 cells, the suppression of MIF by 15d-PGJ2 was weakened; 15d-PGJ2 promoted PPARγ activation in AML 12 cells and primary hepatocytes. Furthermore, the conditioned medium of recombinant MIF- and lipopolysaccharide-treated AML12 respectively promoted BMM migration and inflammatory cytokine expression. Conditioned medium of 15d-PGJ2- or siMif-treated injured AML12 suppressed these effects. Collectively, 15d-PGJ2 activated PPARγ to suppress MIF expression in injured hepatocytes, reducing BMM infiltration and pro-inflammatory activation, ultimately alleviating acute liver injury.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas , Fatores Inibidores da Migração de Macrófagos , Prostaglandina D2 , Animais , Camundongos , Meios de Cultivo Condicionados , Hepatócitos , Fígado , Fatores Inibidores da Migração de Macrófagos/metabolismo , PPAR gama , Prostaglandina D2/uso terapêutico , Prostaglandina D2/farmacologia , Prostaglandinas , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológicoRESUMO
Lily Fusarium wilt disease caused by Fusarium spp. spreads rapidly and is highly destructive, leading to a severe reduction in yield. In this study, lily (Lilium brownii var. viridulum) bulbs were irrigated after planting with suspensions of two Bacillus strains that effectively control lily Fusarium wilt disease to assess their effects on the rhizosphere soil properties and microbial community. A high-throughput sequencing of microorganisms in the rhizosphere soil was performed and the soil physicochemical properties were measured. The FunGuild and Tax4Fun tools were used for a functional profile prediction. The results showed that Bacillus amyloliquefaciens BF1 and B. subtilis Y37 controlled lily Fusarium wilt disease with control efficacies of 58.74% and 68.93%, respectively, and effectively colonized the rhizosphere soil. BF1 and Y37 increased the bacterial diversity and richness of the rhizosphere soil and improved the physicochemical properties of the soil, thereby favoring the proliferation of beneficial microbes. The relative abundance of beneficial bacteria was increased and that of pathogenic bacteria was decreased. Bacillus abundance in the rhizosphere was positively correlated with most soil physicochemical properties, whereas Fusarium abundance was negatively correlated with most physicochemical properties. Functional prediction revealed that irrigation with BF1 and Y37 significantly upregulated glycolysis/gluconeogenesis among metabolism and absorption pathways. This study provides insights into the mechanism by which two Bacillus strains with antifungal activity, BF1 and Y37, antagonize plant pathogenic fungi and lays the foundation for their effective application as biocontrol agents.
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Hepatic macrophages play a central role in liver fibrosis. Scar-associated macrophages (SAMs), a recently identified subgroup of macrophages, play an important role in this process. However, the mechanism by which SAMs transform during liver fibrosis is still unclear. In this study, we aimed to characterize SAMs and elucidate the underlying mechanism of SAM transformation. Bile duct ligation (BDL) and carbon tetrachloride (CCl4) were used to induce mouse liver fibrosis. Non-parenchymal cells were isolated from normal/fibrotic livers and were analyzed using single cell RNA sequencing (scRNA-seq) or mass cytometry (CyTOF). The glucan-encapsulated siRNA particles (siRNA-GeRPs) was employed to perform macrophage selective gene knockdown. The results of scRNA-seq and CyTOF revealed that SAMs, which derived from bone marrow-derived macrophages (BMMs), accumulated in mouse fibrotic livers. Further analysis showed that SAMs highly expressed genes related to fibrosis, indicating the pro-fibrotic functions of SAMs. Moreover, plasminogen receptor Plg-RKT was highly expressed by SAMs, suggesting the role of Plg-RKT and plasminogen (PLG) in SAM transformation. In vitro, PLG-treated BMMs transformed into SAMs and expressed SAM functional genes. Knockdown of Plg-RKT blocked the effects of PLG. In vivo, selective knockdown of Plg-RKT in intrahepatic macrophages of BDL- and CCl4-treated mice reduced the number of SAMs and alleviated BDL- and CCl4-induced liver fibrosis, suggesting that Plg-RKT-PLG played an important role in liver fibrosis by mediating SAM transformation. Our findings reveal that SAMs are crucial participants in liver fibrosis. Inhibition of SAM transformation by blocking Plg-RKT might be a potential therapeutic target for liver fibrosis.
Assuntos
Cicatriz , Plasminogênio , Camundongos , Animais , Plasminogênio/genética , Cicatriz/patologia , Análise da Expressão Gênica de Célula Única , Receptores de Superfície Celular/genética , Macrófagos/patologia , Cirrose Hepática/genética , Cirrose Hepática/patologia , FibroseRESUMO
In this study, cellulose nanofibril (CNF) films from ramie fibers were prepared with different pectin compositions and contents, and the influence of residual pectin on the overall performances of CNF films was evaluated. There was no significant effect of the residual pectin composition on the properties of obtained CNF films. However, when the content of residual pectin was increased from 0.45 % to 9.16 %, the surface area and water absorption of CNF films were increased from 0.2223 to 0.3300 m2/g, and from 93.51 % to 122.42 %, respectively. Pectin covers the CNF surface and act as a physical barrier between the cellulose fibrils; thus the nanocellulose films with high pectin content will have a loose and porous structure, resulting in a high surface area and a high water absorption. Besides, with the residual pectin content decreasing from 9.16 % to 0.45 %, the UVA light transmittance and tensile strength of CNF films were increased from 30.6 % to 59.9 %, and from 37.67 to 100.26 MPa, respectively. After removal of amorphous pectins in CNFs, the low pectin containing CNFs are able to pack more compactly to form a strong and thin film. This paper provides guidance for the preparation of CNF films with different performance requirements.
Assuntos
Boehmeria , Nanofibras , Nanofibras/química , Pectinas , Celulose/química , ÁguaRESUMO
In this study, a series of pretreatment methods (pectin extraction) were employed to prepare cellulose nanofibrils (CNFs) with different pectin composition and contents from ramie fibers. The comprehensive effects of residual pectin on the final properties of CNFs were investigated. The residual pectin did not have a significant effect on the size distribution of the obtained CNFs. While the presence of pectin led to higher zeta potential value, improved dispersion stability and enhanced storage modulus of the CNF dispersion, with the sodium carbonate extraction method showing the greatest impact. The possible mechanism for enhanced dispersion stability of CNFs is the formation of self-assembled hierarchical pectin-hemicellulose/lignin-cellulose nanostructures, offering abundant electrostatic repulsion and steric hindrance between the nanoparticles. This work provides guidelines for the tailored production of CNFs to meet the requirements for different applications.
Assuntos
Boehmeria , Nanofibras , Celulose/química , Lignina/química , Nanofibras/química , PectinasRESUMO
Deep eutectic solvents (DESs), featured as promising green solvents, were applied to examine their effectiveness in pretreating raw ramie fibers (RFs) for cellulose nanofibril (CNF) production. The pretreatment performance of three DESs, i.e., choline chloride-urea (CU), choline chloride-oxalic acid dihydrate (CO), and choline chloride-glycerol (CG), was evaluated based on chemical composition analysis and structural and morphological changes. CO attained the most dramatic morphological changes of RFs, followed by CG and CU. Its high structural disruption of RFs during the pretreatment process, shown in the results from scanning electron microscopy and atomic force microscopy, could be due to an outstanding ability to remove amorphous cellulose and noncellulosic components from raw RFs, confirmed by the results of chemical composition analysis, Fourier transform infrared spectroscopy, and X-ray diffractometry. Overall, this study provided an innovative and effective pretreatment process for fractionating raw cellulosic fibers, so as to promote the subsequent preparation of CNFs.
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Amino acid-based ionic liquids (AAILs) are generally thought of as green solvents and widely used in many regions without systematic assessment of their effect on the environment or human health. In this work, a series of AAILs with different cations and amino acid anions were prepared and characterized, after which their microbial toxicity, phytotoxicity, and biodegradability were evaluated. The results showed that not all AAILs had low toxicity against microorganisms and that some AAILs were highly toxic towards the targeted microorganisms. The phytotoxic effect of the AAILs on rice (Oryza sativa L.) further demonstrated that AAILs should not be presumed to be non-toxic to plants. Moreover, the biodegradability tests showed that majority of AAILs were not satisfactorily biodegradable. In summary, not all AAILs are non-toxic or biodegradable, and their effect on the environment and human health must be assessed before their mass preparation and application.
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Hepatocytes are the main parenchymal cells of the liver and play important roles in liver homeostasis and disease process. The heterogeneity of normal hepatocytes has been reported, but there is little knowledge about hepatocyte subtype and distinctive functions during liver cholestatic injury. Bile duct ligation (BDL)-induced mouse liver injury model was employed, and single-cell RNA sequencing was performed. Western blot and qPCR were used to study gene expression. Immunofluoresence was employed to detect the expressions of marker genes in hepatocytes. We detected a specific hepatocyte cluster (BDL-6) expressing extracellular matrix genes, indicating these hepatocytes might undergo epithelia-mesenchymal transition. Hepatocytes of BDL-6 also performed tissue repair functions (such as angiogenesis) during cholestatic injury. We also found that four clusters of cholestatic hepatocytes (BDL-2, BDL-3, BDL-4, and BDL-5) were involved in inflammatory process in different ways. To be specific, BDL-2/3/5 were inflammation-regulated hepatocytes, while BDL-4 played a role in cell chemotaxis. Among these four clusters, BDL-5 was special. because the hepatocytes of BDL-5 were proliferating hepatocytes. Our analysis provided more knowledge of hepatocyte distinctive functions in injured liver and gave rise to future treatment aiming at hepatocytes.
Assuntos
Colestase , Hepatócitos , Fígado , Análise de Célula Única/métodos , Transcriptoma , Animais , Ductos Biliares/cirurgia , Proliferação de Células , Colestase/metabolismo , Colestase/patologia , Matriz Extracelular/metabolismo , Hepatócitos/metabolismo , Hepatócitos/patologia , Inflamação/metabolismo , Ligadura , Fígado/metabolismo , Fígado/patologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Análise de Sequência de RNA/métodosRESUMO
Cytotoxicity studies are important tools for the assessment of the toxicity of ionic liquids (ILs). In the present study, the cytotoxicity of eleven ILs against Spodoptera frugiperda 9 (Sf-9) cell lines were evaluated via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. The effect on cellular morphology, ultrastructural morphology, and nuclear morphology induced by 1-ethyl-3-methylimidazolium bromide ([C2mim][Br]) was studied via inverted light microscopy observation, acridine orange staining, and transmission electron microscope (TEM) analysis, respectively. The effect on cell DNA fragmentation, cell apoptosis and cell cycle induced by [C2mim][Br] was also investigated via DNA agarose gel electrophoresis and flow cytometry analysis, respectively. The results showed that the cytotoxic effect of ILs on Sf-9 cells was related to the IL structures, concentrations, and length of exposure. The morphological features of apoptosis induced by [C2mim][Br] such as cell shrinkage and convolution, apoptotic bodies, pyknosis, and karyorrhesis were observed. All these phenomena confirmed that Sf-9 cells exposed to [C2mim][Br] died via apoptosis. This study complements the current knowledge about the cytotoxic properties of ILs on insect cells and highlights the mechanism by which ILs kill these cells. Furthermore, it provides a basis for further studies on the future applications of ILs as insecticides.