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BACKGROUND: The interplay between Toxoplasma gondii infection and tumor development is intriguing and not yet fully understood. Some studies showed that T. gondii reversed tumor immune suppression, while some reported the opposite, stating that T. gondii infection promoted tumor growth. METHODS: We created three mouse models to investigate the interplay between T. gondii and tumor. Model I aimed to study the effect of tumor growth on T. gondii infection by measuring cyst number and size. Models II and III were used to investigate the effect of different stages of T. gondii infection on tumor development via flow cytometry and bioluminescent imaging. Mouse strains (Kunming, BALB/c, and C57BL/6J) with varying susceptibilities to tumors were used in the study. RESULTS: The size and number of brain cysts in the tumor-infected group were significantly higher, indicating that tumor presence promotes T. gondii growth in the brain. Acute T. gondii infection, before or after tumor cell introduction, decreased tumor growth manifested by reduced bioluminescent signal and tumor size and weight. In the tumor microenvironment, CD4+ and CD8+ T cell number, including their subpopulations (cytotoxic CD8+ T cells and Th1 cells) had a time-dependent increase in the group with acute T. gondii infection compared with the group without infection. However, in the peripheral blood, the increase of T cells, including cytotoxic CD8+ T cells and Th1 cells, persisted 25 days after Lewis lung carcinoma (LLC) cell injection in the group with acute T. gondii. Chronic T. gondii infection enhanced tumor growth as reflected by increase in tumor size and weight. The LLC group with chronic T. gondii infection exhibited decreased percentages of cytotoxic CD8+ T cells and Th1 cells 25 days post-LLC injection as compared with the LLC group without T. gondii infection. At week 4 post-LLC injection, chronic T. gondii infection increased tumor formation rate [odds ratio (OR) 1.71] in both KM and BALB/c mice. CONCLUSIONS: Our research elucidates the dynamics between T. gondii infection and tumorigenesis. Tumor-induced immune suppression promoted T. gondii replication in the brain. Acute and chronic T. gondii infection had opposing effects on tumor development.
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Modelos Animais de Doenças , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Toxoplasma , Animais , Camundongos , Toxoplasma/imunologia , Toxoplasmose/imunologia , Toxoplasmose/parasitologia , Feminino , Linfócitos T CD8-Positivos/imunologia , Encéfalo/parasitologia , Encéfalo/patologia , Doença Crônica , Microambiente Tumoral , Neoplasias/parasitologia , Doença AgudaRESUMO
BACKGROUND: The inflammatory cascade mediated by macrophages and T cells is considered to be an important factor in promoting the progression of rheumatoid arthritis (RA). Our previous study found that berberine (BBR) can therapeutically impact adjuvant arthritis (AA) in rats through the regulation of macrophage polarization and the balance of Th17/Treg. However, whether BBR's effects on CD4+T cells response are related to its suppression of M1 macrophage still unclear. PURPOSE: The study aimed to estimate the mechanism of BBR in regulating the immunometabolism and differentiation of CD4+T cells are related to exosome derived from M1-macrophage (M1-exo). STUDY-DESIGN/METHODS: Mice model of collagen-induced arthritis (CIA) was established to investigate the antiarthritic effect of BBR was related with regulation of M1-exo to balance T cell subsets. Bioinformatics analysis using the GEO database and meta-analysis. In vitro, we established the co-culture system involving M1-exo and CD4+ T cells to examine whether BBR inhibits CD4+T cell activation and differentiation by influencing M1-exo-miR155. Exosome was characterized using transmission electron microscopy and western blot analysis, macrophage and CD4+T cell subpopulation were detected by flow cytometry. Further, the metabolic profiles of CD4+T cells were assessed by ECAR, OCR, and the level of glucose, lactate, intracellular ATP. RESULT: BBR reinstates CD4+ T cell homeostasis and reduces miR155 levels in both M1-exo and CD4+ T cells obtained from mice with CIA. In vitro, we found exosomes are indispensable for M1-CM on T lymphocyte activation and differentiation. BBR reversed M1-exo facilitating the activation and differentiation of CD4+T cells. Furthermore, BBR reversed glycolysis reprogramming of CD4+T cells induced by M1-exo, while these regulation effects were significantly weakened by miR155 mimic. CONCLUSION: The delivery of miR-155 by M1-exo contributes to CD4+ T cell immunometabolism dysfunction, a process implicated in the development of RA. The anti-arthritic effect of BBR is associated with the suppression of glycolysis and the disruption of CD4+ T cell subsets balance, achieved by reducing the transfer of M1-exo-miR155 into T cells.
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Artrite Experimental , Artrite Reumatoide , Berberina , MicroRNAs , Animais , Camundongos , Ratos , Artrite Experimental/tratamento farmacológico , Artrite Experimental/metabolismo , Artrite Reumatoide/metabolismo , Berberina/farmacologia , Linfócitos T CD4-Positivos , Modelos Animais de Doenças , Macrófagos , MicroRNAs/metabolismoRESUMO
Dysfunction of macrophage polarization majorly contributes to the progression of rheumatoid arthritis (RA). Polarization and functions of activated macrophages are closely associated with the reprogramming of intracellular metabolisms. Previously, we demonstrated that the anti-arthritis effect of berberine (BBR) in rats with adjuvant-induced arthritis (AA) may be related to AMP-activated protein kinase (AMPK) activation (a key regulator in the biological energy metabolism), and balanced macrophage polarization. However, the specific molecular mechanism of BBR in macrophage metabolism is yet to be elucidated. In this study, we clarified that BBR ameliorated articular inflammation and restored M1/M2 ratio in collagen-induced arthritis (CIA) mice in an AMPK-dependent manner. Mechanistically, BBR reversed the effects of mTORC1 agonist leucine (Leu) on regulating macrophage polarization through activation of AMPK to switch glycolytic reprogramming. Furthermore, BBR inhibition of mTORC1 rely on activation of AMPK to phosphorylate raptor and TSC2 instead of destroying its structure. Our study revealed that the activation of AMPK is required for the BBR-mediated anti-arthritis effect by downregulating mTORC1/HIF-1α and inhibiting the glycolysis in M1 macrophages.
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Artrite Experimental , Berberina , Camundongos , Ratos , Animais , Artrite Experimental/tratamento farmacológico , Artrite Experimental/metabolismo , Proteínas Quinases Ativadas por AMP/metabolismo , Berberina/farmacologia , Berberina/uso terapêutico , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Macrófagos , GlicóliseRESUMO
CONTEXT: Ginsenoside metabolite compound K (CK) is an active metabolite produced by ginsenosides in vivo that has an anti-arthritic effect related to the glucocorticoid receptor (GR). However, the potential mechanisms of CK remain unclear. OBJECTIVE: This study explores the role and potential mechanisms of CK in vivo and in vitro. MATERIALS AND METHODS: Adjuvant arthritis (AA) model was induced in Sprague-Dawley (SD) rats; the rats were randomly divided into four groups (n = 10): normal, AA, CK (80 mg/kg), and dexamethasone (Dex) group (1 mg/kg). From day 15, rats were treated with CK (once a day, i.g.) and Dex (once every 3 days, i.p.) for 18 days. To further verify the mechanism of CK, fibroblast-like synoviocytes (FLS) were stimulated by tumour necrosis factor α (TNF-α) to establish an inflammatory model in vitro. RESULTS: CK (80 mg/kg) reduced paw swelling (52%) and arthritis global assessment (31%) compared to that in AA rats. In addition, CK (80 mg/kg) suppressed GLUT1 (38%), HK2 (50%), and PKM2 (56%) levels compared with those in AA FLS. However, the effects of CK (30 µM) on these events were weakened or enhanced after GR knockdown or overexpression in FLS stimulated by TNF-α (30 ng/mL). CK (80 mg/kg) also downregulated the expression of P65 (61%), p-IκB (92%), and HIF-1α (59%). DISCUSSION AND CONCLUSIONS: The inhibition of CK on glycolysis and the NF-κB/HIF-1α pathway is potentially mediated through activating GR. These findings provide experimental evidence for elucidating the molecular mechanism of CK in treating rheumatoid arthritis (RA).
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Artrite Experimental , Ginsenosídeos , Sinoviócitos , Ratos , Animais , NF-kappa B/metabolismo , Receptores de Glucocorticoides/metabolismo , Ginsenosídeos/farmacologia , Artrite Experimental/patologia , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/metabolismo , Proliferação de Células , Fibroblastos/metabolismo , Glicólise , Membrana Sinovial/metabolismoRESUMO
The clinical transformation of polysaccharide-based nano-prodrugs remains a long way off, due to the shackles on easy metabolic clearance, dilemma of dose-dependent toxicity and immunogenicity, and poor tumor selectivity. To address these challenges, the fluorinated dual-crosslinked carboxymethyl chitosan (CMCS)-based nano-prodrugs with precise structure were facilely developed through the reaction of CMCS with water-soluble stimuli-responsive synergistic small molecule prodrug (Pt(IV)-1), glutaraldehyde and heptafluorobutyric anhydride successively. The fluorination enabled the nano-prodrugs to display metabolic stability and improve tumoral cellular uptake. The pH/glutathione (GSH)-sensitive dual-crosslinked structure enabled the nano-prodrugs to show physicochemical stability at physiological pH, selective drug release and synergistic cytotoxicity at tumoral intracellular pH/GSH, and circumventing the dilemma of dose-dependent toxicity and immunogenicity induced by that crosslinked or grafted via a single drug. These superior performances promoted stability in long-term storage and circulation, normal blood routine and aminotransferase, fantastic hemocompatibility, selective tumor accumulation and precisely synergistic chemotherapy, therefore achieving significant tumor growth inhibition while minimizing side effects. Thus, the precise fluorinated dual-crosslinked CMCS-based nano-prodrugs have great potential for selective clinical cancer treatment.
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Antineoplásicos , Quitosana , Nanopartículas , Neoplasias , Pró-Fármacos , Humanos , Pró-Fármacos/química , Quitosana/química , Nanopartículas/química , Portadores de Fármacos/química , Antineoplásicos/farmacologia , Antineoplásicos/químicaRESUMO
BACKGROUND: Toxoplasma gondii, an intracellular protozoan parasite, exists in the host brain as cysts, which can result in Toxoplasmic Encephalitis (TE) and neurological diseases. However, few studies have been conducted on TE, particularly on how to prevent it. Previous proteomics studies have showed that the expression of C3 in rat brains was up-regulated after T. gondii infection. METHODS: In this study, we used T. gondii to infect mice and bEnd 3 cells to confirm the relation between T. gondii and the expression of C3. BEnd3 cells membrane proteins which directly interacted with C3a were screened by pull down. Finally, animal behavior experiments were conducted to compare the differences in the inhibitory ability of TE by four chemotherapeutic compounds (SB290157, CVF, NSC23766, and Anxa1). RESULTS: All chemotherapeutic compounds in this study can inhibit TE and cognitive behavior in the host. However, Anxa 1 is the most suitable material to inhibit mice TE. CONCLUSION: T. gondii infection promotes TE by promoting host C3 production. Anxa1 was selected as the most appropriate material to prevent TE among four chemotherapeutic compounds closely related to C3.
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Toxoplasma , Toxoplasmose Cerebral , Animais , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Camundongos , Proteômica , Toxoplasmose Cerebral/tratamento farmacológico , Toxoplasmose Cerebral/metabolismo , Toxoplasmose Cerebral/parasitologiaRESUMO
BACKGROUND: In order to avoid excessive treatment of thyroid nodules in the clinic, it is necessary to find a simple and practical analysis method to comprehensively and accurately reflect benign or malignant thyroid nodules. This study aimed to construct and validate a comprehensive and reliable network-based predictive model using a variety of imaging and laboratory criteria for thyroid nodules to stratify the risk of malignancy prior to surgery. METHODS: We retrospectively analyzed data from patients who underwent surgical treatment for thyroid nodules at the Thyroid and Breast Diagnosis and Treatment Center of Weifang Hospital of Traditional Chinese Medicine between January 2018 and December 2020. Binary logical regression analysis was performed to predict whether nodules were malignant or benign. The developmental dataset included 457 patients (January 2018-December 2020). The validation set included separate data points (n = 225, January 2018-December 2020). RESULTS: In this study, criteria that showed significant predictive value for malignant nodules included TI-RADS: 4b (p = 0.065); Bethesda IV, Bethesda V, Bethesda VI (P < 0.0001); BRAFV600E mutation (P < 0.0001); Calcitonin>5 pg/ml (p = 0.0037); and FNA-Tg>30 ng/ml (p = 0.0003). A 10-grade risk scoring system was developed. The risk of malignancy risk ranged from 2.06% to 100% and was positively associated with increasing risk grade. The areas under the receiver-operating characteristic curve of the development and validation sets were 0.972 and 0.946, respectively. CONCLUSION: A simple, comprehensive and reliable web-based predictive model was designed using a variety of imaging and laboratory criteria to stratify thyroid nodules by probability of malignancy.
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Neoplasias da Glândula Tireoide , Nódulo da Glândula Tireoide , Humanos , Curva ROC , Estudos Retrospectivos , Neoplasias da Glândula Tireoide/patologia , Nódulo da Glândula Tireoide/diagnóstico por imagem , Nódulo da Glândula Tireoide/cirurgia , Ultrassonografia/métodosRESUMO
To achieve highly selective synergistic chemotherapy attractive for clinical translation, the precise polymeric nano-prodrugs (PPD-NPs) were successfully constructed via the facile crosslinking reaction between pH-sensitive poly(ortho ester)s and reduction-sensitive small molecule synergistic prodrug (Pt(IV)-1). PPD-NPs endowed the defined structure and high drug loading of cisplatin and demethylcantharidin (DMC). Moreover, PPD-NPs exhibited steady long-term storage and circulation via the crosslinked structure, suitable negative potentials and low critical micelle concentration (CMC), improved selective tumour accumulation and cellular internalization via dynamic size transition and surficial amino protonation at tumoural extracellular pH, promoted efficient disintegration and drug release at tumoural intracellular pH/glutathione, and enhanced cytotoxicity via the synergistic effect between cisplatin and DMC with the feed ratio of 1:2, achieving significant tumour suppression while decreasing the side effects. Thus, the dynamic crosslinked polymeric nano-prodrugs exhibit tremendous potential for clinically targeted synergistic cancer therapy.
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Macrophages are highly plastic cells critical for the development of rheumatoid arthritis (RA). Macrophages exhibit a high degree of pro-inflammatory plasticity in RA, accompanied by a metabolic reprogramming from oxidative phosphorylation (OXPHOS) to glycolysis. 2-deoxyglucose (2-DG), a glycolysis inhibitor, has previously been shown to exhibit anti-inflammatory and anti-arthritic properties. However, the specific mechanisms of inflammatory modulation by 2-DG remain unclear. This study used 2-DG to treat rats with adjuvant arthritis (AA) and investigated its specific anti-arthritic mechanisms in the murine-derived macrophage cell line RAW264.7 in vitro. 2-DG reduced the arthritis index as well as alleviated cellular infiltration, synovial hyperplasia, and bone erosion in AA rats. Moreover, 2-DG treatment modulated peritoneal macrophage polarization, increasing levels of the arginase1 (Arg1) and decreasing expression of the inducible nitric oxide synthase (iNOS). 2-DG activated AMP-activated protein kinase (AMPK) via phosphorylation and reduced activation of the nuclear factor κB (NF-κB) in peritoneal macrophages of AA rats. In vitro, we verified that 2-DG promoted macrophage transition from M1 to M2-type by upregulating the expression of p-AMPKα and suppressing NF-κB activation in LPS-stimulated RAW264.7 cells. LPS-induced macrophages exhibited a metabolic shift from glycolysis to OXPHOS following 2-DG treatment, as observed by reduced extracellular acidification rate (ECAR), lactate export, glucose consumption, as well as an elevated oxygen consumption rate (OCR) and intracellular ATP concentration. Importantly, changes in polarization and metabolism in response to 2-DG were dampened after AMPKα knockdown. These findings indicate that the anti-arthritic 2-DG effect is mediated by a modulation of macrophage polarization in an AMPK-dependent manner.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Artrite Experimental/patologia , Polaridade Celular , Desoxiglucose/farmacologia , Glicólise/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/patologia , Animais , Artrite Experimental/enzimologia , Movimento Celular/efeitos dos fármacos , Polaridade Celular/efeitos dos fármacos , Modelos Animais de Doenças , Ativação Enzimática/efeitos dos fármacos , Inflamação/patologia , Articulações/patologia , Lipopolissacarídeos , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , NF-kappa B/metabolismo , Fagocitose/efeitos dos fármacos , Células RAW 264.7 , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacosRESUMO
Taenia solium cysts were collected from pig skeletal muscle and analyzed via a shotgun proteomic approach to identify known proteins in the cyst fluid and to explore host-parasite interactions. Cyst fluid was aseptically collected and analyzed with shotgun liquid chromatography-tandem mass spectrometry (LC-MS/MS). Gene alignment and annotation were performed using Blast2GO software followed by gene ontology analysis of the annotated proteins. The pathways were further analyzed with the Kyoto Encyclopedia of Genes and Genomes (KEGG), and a protein-protein interaction (PPI) network map was generated using STRING software. A total of 158 known proteins were identified, most of which were low-molecular-mass proteins. These proteins were mainly involved in cellular and metabolic processes, and their molecular functions were predominantly related to catalytic activity and binding functions. The pathway enrichment analysis revealed that the known proteins were mainly enriched in the PI3K-Akt and glycolysis/gluconeogenesis signaling pathways. The nodes in the PPI network mainly consisted of enzymes involved in sugar metabolism. The cyst fluid proteins screened in this study may play important roles in the interaction between the cysticerci and the host. The shotgun LC-MS/MS, gene ontology, KEGG, and PPI network map data will be used to identify and analyze the cyst fluid proteome of cysticerci, which will provide a basis for further exploration of the invasion and activities of T. solium.
Assuntos
Proteínas de Helminto/análise , Proteômica/métodos , Taenia solium/química , Animais , Cromatografia Líquida , Proteínas de Helminto/classificação , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Interações Hospedeiro-Parasita , Anotação de Sequência Molecular/métodos , Peso Molecular , Músculo Esquelético/parasitologia , Mapas de Interação de Proteínas , Alinhamento de Sequência , Transdução de Sinais , Suínos , Taenia solium/genética , Espectrometria de Massas em TandemRESUMO
BACKGROUND: The life cycle of Taenia solium is characterized by different stages of development, requiring various kinds of hosts that can appropriately harbor the eggs (proglottids), the oncospheres, the larvae and the adults. Similar to other metazoan pathogens, T. solium undergoes transcriptional and developmental regulation via epigenetics during its complex lifecycle and host interactions. RESULT: In the present study, we integrated whole-genome bisulfite sequencing and RNA-seq technologies to characterize the genome-wide DNA methylation and its effect on transcription of Cysticercus cellulosae of T. solium. We confirm that the T. solium genome in the cysticercus stage is epigenetically modified by DNA methylation in a pattern similar to that of other invertebrate genomes, i.e., sparsely or moderately methylated. We also observed an enrichment of non-CpG methylation in defined genetic elements of the T. solium genome. Furthermore, an integrative analysis of both the transcriptome and the DNA methylome indicated a strong correlation between these two datasets, suggesting that gene expression might be tightly regulated by DNA methylation. Importantly, our data suggested that DNA methylation might play an important role in repressing key parasitism-related genes, including genes encoding excretion-secretion proteins, thereby raising the possibility of targeting DNA methylation processes as a useful strategy in therapeutics of cysticercosis.
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Metilação de DNA , Genoma Helmíntico , Taenia solium/genética , Animais , Epigenômica , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , RNA-Seq , Sequenciamento Completo do GenomaRESUMO
Berberine (BBR) is the effective constituent of Cortex phellodendri and was characterized as an excellent anti-microbial agent with significant anti-inflammatory effects. Previously, we had demonstrated that BBR alleviated the inflammatory response in adjuvant-induced arthritis (AA) rats by regulating polarization of macrophages. However, the exact mechanics by which BBR regulates macrophage polarization remained unclear. Here, we showed that BBR treatment had little influence on total number of macrophages in joints of AA rats, but increased the proportion of M2 macrophages and decreased the proportion of M1 macrophages. Meanwhile, we found BBR up-regulated the expression of AMP-activated protein kinase phosphorylation (p-AMPK) and down-regulated the expression of Hypoxia inducible factor 1α (HIF-1α) in synovial macrophages of AA rats. In vitro, using LPS-stimulated peritoneal macrophages from normal rats, we also verified that pretreatment with BBR promoted transition from M1 to M2 by up-regulating the expression of p-AMPK and suppressing the expression of HIF-1α. Compound C (an AMPK inhibitor) could abrogate the inhibition of BBR on migration of macrophages. Glycolysis of M1 suppressed by BBR through decreasing lactate export, glucose consumption, and increasing intracellular ATP content, which was remarkably reversed by Compound C. These findings indicated that anti-arthritis effect of BBR is associated with regulating energy metabolism of macrophages through AMPK/HIF-1α pathway.
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Anti-Inflamatórios/uso terapêutico , Artrite Experimental/tratamento farmacológico , Berberina/uso terapêutico , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Articulação do Tornozelo/efeitos dos fármacos , Articulação do Tornozelo/imunologia , Articulação do Tornozelo/patologia , Anti-Inflamatórios/farmacologia , Artrite Experimental/imunologia , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Berberina/farmacologia , Citocinas/sangue , Metabolismo Energético/efeitos dos fármacos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Ratos Sprague-DawleyRESUMO
The present study primarily aims to examine differences in the use of tobacco and alcohol by junior high school students under different parental control levels (including parental psychological control and parental behavioral control). It thus explores the regulatory effect of parental control on the relationship between adolescent sensation seeking and tobacco and alcohol use. A total of 1,050 junior high school students in Shandong province were surveyed using sensation-seeking scale, parental control scale, and adolescent health-related risk behavior questionnaire. As the results showed, (1) sensation seeking and gender had effects on the use of tobacco and alcohol among junior high school students; (2) parental psychological control can enhance and moderate the relationship between sensation seeking and the use of tobacco and alcohol; (3) parental behavioral control cannot regulate the relationship between sensation seeking and the use of tobacco and alcohol among junior high school students; and (4) no significant urban-rural differences were found regarding the regulatory effects of parental psychological control on sensation seeking and alcohol and tobacco use in junior high school students.
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Berberine (BBR) is a traditional folk medicine with excellent anti-inflammatory properties. This study aimed to investigate the anti-arthritic effects of BBR in adjuvant arthritis (AA) in rats and its regulatory role in the polarization of macrophages. Rats were immunized with Complete Freund's Adjuvant (CFA), and then BBR (40, 80, 160â¯mg/kg) was administered orally for 14 days. BBR significantly reduced paw swelling and arthritis global assessment as well as alleviated joint destruction and inflammatory cell infiltration. The index of the thymus and thymocyte proliferation were significantly reduced by BBR. Moreover, BBR treatment restrained the phagocytic function of macrophages and restored the balance of M1/M2 by reducing the levels of M1 cytokines (tumour necrosis factor-α, interleukin-1ß, and interleukin-6), increasing the levels of M2 cytokines (interleukin-10 and transforming growth factor-ß1), increasing the expression of arginase 1(Arg1) (M2 marker) and decreasing the expression of inducible nitric oxide synthase (iNOS) (M1 marker). BBR also downregulated the ratio of Th17/Treg cells. Further research on the adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK)/nuclear factor κB (NF-κB) pathway found that BBR upregulated the activity of AMPK, while it downregulated the expression of phospho-RelA (p-p65), phospho-NF-kappa-B inhibitor alpha (p-IκBα) and cyclooxygenase (COX)-2. Therefore, our findings suggest BBR has significantly therapeutic effects in AA rats by regulating the polarization of macrophages through the AMPK/NF-кB pathway.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Artrite Experimental/tratamento farmacológico , Berberina/farmacologia , Fatores Imunológicos/farmacologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , NF-kappa B/metabolismo , Animais , Articulação do Tornozelo/diagnóstico por imagem , Articulação do Tornozelo/patologia , Arginase/metabolismo , Artrite Experimental/imunologia , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Berberina/uso terapêutico , Contagem de Células , Proliferação de Células/efeitos dos fármacos , Citocinas/biossíntese , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Fatores Imunológicos/uso terapêutico , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Óxido Nítrico Sintase Tipo II/metabolismo , Fagocitose/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacos , Linfócitos T Reguladores/citologia , Linfócitos T Reguladores/efeitos dos fármacos , Células Th17/citologia , Células Th17/efeitos dos fármacos , Tomografia Computadorizada por Raios XRESUMO
OBJECTIVE: To observe the effects of 2-deoxyglucose inhibiting synovial pannus of adjuvant arthritis rats and to explore its potential mechanism of inhibiting angiogenesis by investigating proliferation, migration and matrigel tube formation assay in vitro. METHODS: The effect of 2-DG on synovial pannus was evaluated by histopathology of HE staining; HUVEC proliferation was determined by CCK-8 method; migration of FLS were determined by transwell; In vitro matrigel tube formation assay was made for assessing tube number of HUVEC; p-AMPK and Bcl-2 were detected by Western blot assay; AMPK signaling pathway in HUVEC was inhibited by compound C, which is an inhibitor of AMPK activation. RESULTS: 2-DG (200 mg/kg) obviously decreased appearance of synovial pannus (P < 0.01); in vitro, 2-DG (0.5 mmol/L and/or 5 mmol/L) obviously inhibited proliferation, migration and tube number of HUVEC (P < 0.01 or P < 0.001), and its effects on HUVEC were reversed by using AMPK antagonist (Compound C); Western blot showed that 2-DG (5 mmol/L) increased expression of p-AMPK and decreased expression of Bcl-2 (P < 0.05). CONCLUSIONS: Activating AMPK pathway and decreasing expression of Bcl-2 may the potential mechanism by which 2-DG contributes to anti-angiogenesis and effects of inhibiting proliferation, migration and tube number of HUVEC.
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Inibidores da Angiogênese/farmacologia , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Desoxiglucose/farmacologia , Neovascularização Patológica/prevenção & controle , Proteínas Quinases/metabolismo , Membrana Sinovial/efeitos dos fármacos , Quinases Proteína-Quinases Ativadas por AMP , Animais , Artrite Experimental/patologia , Artrite Reumatoide/patologia , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Ratos , Membrana Sinovial/patologiaRESUMO
A specific and sensitive liquid chromatography-tandem mass spectrometric method for quantitative determination of paclitaxel in rat plasma was developed and validated using docetaxel as an internal standard. Liquid-liquid extraction using tert-butyl methyl ether was used to extract the drug and the internal standard from plasma. The separation of paclitaxel was performed on a C(18) column with a mobile phase of acetonitrile:water:formic acid (65:35:0.1, v/v/v) over 5min. The assay was based on the selected reaction monitoring transitions at m/z of the precursor-product ion transitions m/z 854.2-->286.1 for paclitaxel and 808.3-->527.2 for internal standard. The lower limit of quantification was 0.5ng/mL based on 100microL of plasma. Intra- and inter-day assay variations were less than 15%, and the accuracy values were between 95.4 and 105.4%. The extraction recoveries ranged from 96.7 to 103.7% across the calibration curve range. The method was successfully applied to measurement of low concentrations of paclitaxel or regenerated paclitaxel in plasma after intravenous administration of a single dose (10mg/kg) of a poly(l-glutamic acid)-alanine-paclitaxel conjugate to rats.
Assuntos
Alanina/sangue , Antineoplásicos Fitogênicos/sangue , Cromatografia Líquida , Paclitaxel/análogos & derivados , Ácido Poliglutâmico/análogos & derivados , Espectrometria de Massas em Tandem , Alanina/administração & dosagem , Alanina/análogos & derivados , Alanina/farmacocinética , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/farmacocinética , Química Farmacêutica , Cromatografia Líquida/normas , Estabilidade de Medicamentos , Injeções Intravenosas , Masculino , Paclitaxel/administração & dosagem , Paclitaxel/sangue , Paclitaxel/farmacocinética , Ácido Poliglutâmico/administração & dosagem , Ácido Poliglutâmico/sangue , Ácido Poliglutâmico/farmacocinética , Ratos , Ratos Sprague-Dawley , Padrões de Referência , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem/normasRESUMO
OBJECTIVE: To analyze the influence of the body mass index (BMI) and age on deep vein thrombosis (DVT) after the total hip arthroplasty (THA) or total knee arthroplasty(TKA) in Chinese patients who received prophylactic treatment for DVT. METHODS: We used a randomized clinical case-control study. From April 2004 to August 2004, we performed THA and TKA for 95 patients (128 hips and knees). There were 27 men and 68 women with an average age of 60 years (range, 23-78) at surgery, and with an average BMI of 25.88 kg/m2 (range, 14.34-40.39) before surgery. All the patients were given low-molecular-weight heparin for 7-10 days pre- and postoperatively to prevent DVT. Color Doppler ultrasonography was used to detect DVT of bilateral lower extremities in all the patients before operation and 7-10 days after operation. The patients were divided into 4 groups according to their BMI:the non-obese group (BMI < or = 25.00 kg/m2), the overweight group (BMI, 25.01-27.00 kg/m2), the obese group (BMI, 27.01-30.00 kg/m2), and the morbidly obese group (BMI > 30.00 kg/m2); and they were also divided into 4 groups according to their ages (< or =40, 41-60, 61-70, >70 yr) so as to analyze the influence of the BMI and age on DVT. RESULTS: The total incidence of DVT in all the patients was 47.4% (45/95) and the incidence of proximal DVT was 3.2%. Forty-five patients had DVT and 50 patients had no DVT. The average BMI of the patients with DVT was significantly higher than that of the patients with no DVT (P < 0.05). The overweight, obese, and morbidly obese patients had an odds ratio of 7.04, 4.8, and 9.6 for DVT compared with the non-obese patients (P < 0.05); but the obese group had a less risk than the other 2 groups. The 41-60, 61-70, and > 70-year-old patients had an odds ratio of 24.0, 38.2, and 24.4 for DVT compared with the < or = 40-year-old patients (P < 0.05). CONCLUSION: Obesity (BMI> 25 kg/m2) and an increasing age (>40 yr) are identified as statistically significant risk factors for DVT after THA or TKA, and patients aged 61-70 years are more likely to have DVT than the other patients. We should pay more attention to those obesity and aged patients when they are undergoing the total joint replacement, and we should give them enough prophylaxis and closely observe the symptoms in their bilateral lower extremities after operation, taking ultrasonography or venography to check DVT if necessary, so that we can give them prompt treatment and prevent fatal pulmonary thromboembolism.
Assuntos
Artroplastia de Quadril/efeitos adversos , Artroplastia do Joelho/efeitos adversos , Trombose Venosa/etiologia , Adulto , Fatores Etários , Idoso , Índice de Massa Corporal , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Obesidade/complicações , Complicações Pós-OperatóriasRESUMO
OBJECTIVE: To analyze the clinical risk factors for deep vein thrombosis (DVT) after total hip and knee arthroplasty in Chinese patients who received prophylactic treatment for DVT. METHODS: We evaluated 128 total hip arthroplasty (THA) and total knee arthroplasty (TKA) in 95 patients performed at our center from April 2004 to August 2004, which included 48 THAs in 43 patients and 80 TKAs in 52 patients. There were 27 men and 68 women with a mean age of 59.77 years (range, 23-78 years). All patients had been given low-molecular-weight heparin before operation and for 7-10 days post-operation to prevent DVT. Color Doppler ultrasonography was used to detect DVT of bilateral lower extremities in all patients before operation and at 7-10 days after operation. Nineteen clinical factors were examined preoperation and 7-10 days post-operation in order to analyze their influences on DVT formation after surgery. RESULTS: There were 45 patients who developed DVT after operation. The incidence of DVT in all patients was 47.4% (45/95) and the incidence of proximal DVT was 3.2%. There were more asymptomatic DVT (57.8%, 26/45) than symptomatic ones, and some patients without DVT (14%, 7/50) presented some of the DVT symptoms. Logistic regression analysis demonstrated a definite association of female, obesity (representative by BMI), cement usage and diagnosed RA with DVT with odds ratio of 10.008, 3.094, 8.887, and 0.194 respectively. Other clinical factors had no statistically significant association with DVT. CONCLUSIONS: Female, obesity, and cement usage were the risk factors for DVT after THA and TKA, and diagnosed RA was the protecting factors for DVT after THA and TKA. Other clinical factors such as age, OA, type of implant, monolateral or bilateral operation, duration of anesthesia, surgery and bandage usage for blood control, time for immobilization et al were not the risk factors for DVT.