RESUMO
Hashimoto's thyroiditis (HT) is the most common organ-specific autoimmune disease, predominantly affecting women. Although the pathogenesis of HT is incompletely understood, some studies have found that macrophage polarization plays a role. Puerarin is a soy isoflavone compound that has anti-inflammatory and immunomodulatory effects and regulates macrophage immune activity. This study aimed to verify the therapeutic effect of puerarin on HT and explored its regulatory effect on macrophage polarization imbalance in HT. Through bioinformatics analysis and molecular biology methods, it was found that macrophages increased significantly in HT patients and model mice. Immunological staining showed that puerarin intervention could reduce tissue inflammatory cell infiltration. Molecular biological examination displayed that puerarin could inhibit local and systemic inflammation levels, and the expression of marker thyroglobulin and thyroid peroxidase Abs. In vivo experimental results indicated that puerarin regulated macrophage polarity and reduced inflammatory damage, possibly by inhibiting the pyroptosis signaling pathway. In vivo macrophage clearance experiments demonstrated that puerarin relied on macrophages to exert its mechanism of action in treating HT. The results of this study indicate that macrophages are important mediators in the development of HT, and puerarin can regulate macrophage polarity and inflammatory status to provide thyroid tissue protection, which provides a new idea for the treatment of HT.
Assuntos
Isoflavonas , Macrófagos , Isoflavonas/farmacologia , Isoflavonas/uso terapêutico , Animais , Camundongos , Macrófagos/imunologia , Macrófagos/efeitos dos fármacos , Humanos , Feminino , Modelos Animais de Doenças , Tireoidite Autoimune/tratamento farmacológico , Tireoidite Autoimune/imunologia , Doença de Hashimoto/tratamento farmacológico , Doença de Hashimoto/imunologia , Ativação de Macrófagos/efeitos dos fármacos , Ativação de Macrófagos/imunologia , Piroptose/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacosRESUMO
Indoleamine-2,3-dioxygenase 1 (IDO1) and signal transducer and activator of transcription 3 (STAT3) have emerged as significant targets in the tumor microenvironment for cancer therapy. In this study, we synthesized three novel 2-amino-1,4-naphthoquinone amide-oxime derivatives and identified them as dual inhibitors of IDO1 and STAT3. The representative compound NK3 demonstrated effective binding to IDO1 and exhibited good inhibitory activity (hIDO1 IC50 = 0.06 µM), leading to its selection for further investigation. The direct interactions between compound NK3 and IDO1 and STAT3 proteins were confirmed through surface plasmon resonance analysis. A molecular docking study of compound NK3 revealed key interactions between NK3 and IDO1, with the naphthoquinone-oxime moiety coordinating with the heme iron. In the in vitro anticancer assay, compound NK3 displayed potent antitumor activity against selected cancer cell lines and effectively suppressed nuclear translocation of STAT3. Moreover, in vivo assays conducted on CT26 tumor-bearing Balb/c mice and an athymic HepG2 xenograft model revealed that compound NK3 exhibited potent antitumor activity with low toxicity relative to 1-methyl-L-tryptophan (1-MT) and doxorubicin (DOX). Overall, these findings provided evidence that the dual inhibitors of IDO1 and STAT3 may offer a promising avenue for the development of highly effective drug candidates for cancer therapy.
Assuntos
Naftoquinonas , Fator de Transcrição STAT3 , Humanos , Animais , Camundongos , Simulação de Acoplamento Molecular , Estudos Prospectivos , Amidas/farmacologia , Camundongos Endogâmicos BALB C , Naftoquinonas/farmacologia , Oximas/farmacologiaRESUMO
Proficient mismatch repair or microsatellite stable (pMMR/MSS) colorectal cancers (CRCs) are vastly outnumbered by deficient mismatch repair or microsatellite instability-high (dMMR/MSI-H) tumors and lack a response to immune checkpoint inhibitors (ICIs). In this study, we reported two distinct expression patterns of ASCL2 in pMMR/MSS and dMMR/MSI-H CRCs. ASCL2 is overexpressed in pMMR/MSS CRCs and maintains a stemness phenotype, accompanied by a lower density of tumor-infiltrating lymphocytes (TILs) than those in dMMR/MSI CRCs. In addition, coadministration of anti-PD-L1 antibodies facilitated T cell infiltration and provoked strong antitumor immunity and tumor regression in the MC38/shASCL2 mouse CRC model. Furthermore, overexpression of ASCL2 was associated with increased TGFB levels, which stimulate local Cancer-associated fibroblasts (CAFs) activation, inducing an immune-excluded microenvironment. Consistently, mice with deletion of Ascl2 specifically in the intestine (Villin-Cre+, Ascl2 flox/flox, named Ascl2 CKO) revealed fewer activated CAFs and higher proportions of infiltrating CD8+ T cells; We further intercrossed Ascl2 CKO with ApcMin/+ model suggesting that Ascl2-deficient expression in intestinal represented an immune infiltrating environment associated with a good prognosis. Together, our findings indicated ASCL2 induces an immune excluded microenvironment by activating CAFs through transcriptionally activating TGFB, and targeting ASCL2 combined with ICIs could present a therapeutic opportunity for MSS CRCs.
Assuntos
Fibroblastos Associados a Câncer , Neoplasias do Colo , Neoplasias Colorretais , Animais , Camundongos , Linfócitos T CD8-Positivos , Neoplasias Colorretais/genética , Modelos Animais de Doenças , Instabilidade de Microssatélites , Repetições de MicrossatélitesRESUMO
A series of chromone-oxime derivatives containing piperazine sulfonamide moieties were designed, synthesized and evaluated for their inhibitory activities against IDO1. These compounds displayed moderate to good inhibitory activity against IDO1 with IC50 values in low micromolar range. Among them, compound 10m bound effectively to IDO1 with good inhibitory activities (hIDO1 IC50 = 0.64 µM, HeLa IDO1 IC50 = 1.04 µM) and were selected for further investigation. Surface plasmon resonance analysis confirmed the direct interaction between compound 10m and IDO1 protein. Molecular docking study of the most active compound 10m revealed key interactions between 10m and IDO1 in which the chromone-oxime moiety coordinated to the heme iron and formed several hydrogen bonds with the porphyrin ring of heme and ALA264, consistent with the observation by UV-visible spectra that 10m induced a Soret peak shift from 403 to 421 nm. Moreover, compound 10m exhibited no cytotoxicity at its effective concentration in MTT assay. Consistently, in vivo assays results demonstrated that 10m displayed potent antitumor activity with low toxicity in CT26 tumor-bearing Balb/c mice, in comparison with 1-methyl-l-tryptophan (1-MT) and 4-amino-N-(3-chloro-4-fluorophenyl)-N'-hydroxy-1,2,5-oxadiazole-3-carboximidamide (IDO5L). In brief, the results suggested that chromone-oxime derivatives containing sulfonamide moieties might serve as IDO1 inhibitors for the development of new antitumor agents.
Assuntos
Inibidores Enzimáticos , Indolamina-Pirrol 2,3,-Dioxigenase , Animais , Camundongos , Relação Estrutura-Atividade , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/química , Simulação de Acoplamento Molecular , Oximas/farmacologia , Heme , Sulfonamidas/farmacologiaRESUMO
By developing a filtering framework and a sector-level multi-regional input-output structural decomposition model, this study identifies key common emission sources, motivation sources, and inter-provincial emission flows of both GHGs and air pollutants and reveals the key driving forces of changes in different emissions from 2012 to 2017. Results show that key common emission sources are electricity sector, non-metallic mineral products, and smelting and processing of metals in Shandong and Hebei. However, key common motivation sources are the construction sectors in Guangdong, Henan, Jiangsu, Zhejiang, and Shandong. The key inflow regions include Guangdong and Zhejiang and key outflow regions include Jiangsu and Hebei. The emission reductions are attributed to the emission intensity effect of the construction sector; contrastingly, the emission increase is from the investment scale of the construction sector. Here, Jiangsu could be a key target for future emission reduction because of its high absolute emissions and low past reduction. The scale of investment in construction might be a significant factor in reducing emissions in Shandong and Guangdong. Henan and Zhejiang could concentrate on sound new building planning and resource recycling.
Assuntos
Poluentes Atmosféricos , Gases de Efeito Estufa , Poluentes Atmosféricos/análise , ChinaRESUMO
Methylprednisolone pulse treatment is currently used for optic neuritis. It can speed visual recovery, but does not improve the ultimate visual outcomes. Recent studies have reported that miR-125a-5p has immunomodulatory effects on autoimmune diseases. However, it remains unclear whether miR-125a-5p has effects on optic neuritis. In this study, we used adeno-associated virus to overexpress or silence miR-125a-5p in mice. We found that silencing miR-125a-5p increased the latency of visual evoked potential and aggravated inflammation of the optic nerve. Overexpression of miR-125a-5p suppressed inflammation of the optic nerve, protected retinal ganglion cells, and increased the percentage of Treg cells. Our findings show that miR-125a-5p exhibits anti-inflammatory effects through promoting the differentiation of Treg cells.
RESUMO
BACKGROUND: Birt-Hogg-Dubé Syndrome (BHDS) characterised by skin fibrofolliculomas, kidney tumour and pulmonary cysts/pneumothorax is caused by folliculin (FLCN) germline mutations. The pathology of both neoplasia and focused tissue loss of BHDS strongly features tissue-specific behaviour of the gene. Isolated cysts/pneumothorax is the most frequent atypical presentation of BHDS and often misdiagnosed as primary spontaneous pneumothorax (PSP). Deferential diagnosis of BHDS with isolated pulmonary presentation (PSP-BHD) from PSP is essential in lifelong surveillance for developing renal cell carcinoma. METHODS: The expression profiles of microRNAs (miRNAs) in cystic lesions of PSP-BHD and PSP were determined via microarray. The selected upregulated miRNAs were further confirmed in the plasma of an expanded cohort of PSP-BHD patients by reverse transcription quantitative PCR (RT-qPCR). Their diagnostic accuracy was evaluated. Moreover, the cellular functions and targeted signalling pathways of FLCN-regulated miRNAs were assessed in various cell lines and in the lesion tissue contexts. RESULTS: Cystic lesions of PSP-BHD and PSP showed different miRNAs profiles with a significant upregulation of miR-424-5p and let-7d-5p in PSP-BHD. The combination of the two effectively predicted BHDS patients. In vitro studies revealed a suppressive effect of FLCN on miR-424-5p and let-7d-5p expressions specifically in lung epithelial cells. The ectopic miRNAs triggered epithelial apoptosis and epithelial transition of mesenchymal cells and suppressed the reparative responses in cells and tissues with FLCN deficiency. CONCLUSION: The upregulation of miR-424-5p and let-7d-5p by FLCN deficiency occurred in epithelial cells and marked the PSP-BHD condition, which contributed to a focused degenerative pathology in the lung of PSP-BHD patients.
Assuntos
Síndrome de Birt-Hogg-Dubé/patologia , Células Epiteliais/patologia , Estrona/metabolismo , Pulmão/patologia , MicroRNAs/metabolismo , Adulto , Apoptose , Síndrome de Birt-Hogg-Dubé/genética , Síndrome de Birt-Hogg-Dubé/metabolismo , Linhagem Celular , Células Cultivadas , China , Diagnóstico Diferencial , Células Epiteliais/metabolismo , Feminino , Humanos , Pulmão/metabolismo , Masculino , Análise Serial de Proteínas , Estudos RetrospectivosRESUMO
RATIONALE: Myiasis is a parasitic disease caused by fly larvae of the Diptera order that infest human and other vertebrate animal tissues. Orbital myiasis is a potentially destructive infestation of the orbital tissues, which may affect individuals with previous ocular diseases or disorders of consciousness. PATIENT CONCERNS: A 72-year-old man presented with a complaint of repeated pain for two years after trauma to his right eyelid and aggravated symptoms with larvae wriggling out for 2 days. An orbital computed tomography scan revealed right eyeball protrusion and periocular soft tissue edema. Two days later, magnetic resonance imaging showed that the shape of the right eyeball was changed and that the normal structure of the eyeball could not be identified. DIAGNOSES: Due to the patient's symptoms and imaging examination results, the diagnosis of orbital myiasis was made. INTERVENTIONS: The patient was treated by exenteration of the right orbit, and all necrotic tissues and larvae were removed. The defect was repaired via reconstruction with a pedicled musculocutaneous flap from the forehead region. Antibiotics and tetanus toxoid therapy were utilized to prevent potential bacterial infection. OUTCOMES: The patient recovered well postoperatively and was discharged uneventfully. During the 6-month follow-up period, the wound healed well. LESSONS: Advanced age and untreated eye trauma are risk factors for orbital myiasis. Timely removal of larvae and elimination of infections are important measures for protecting the eyeball.
Assuntos
Enucleação Ocular/métodos , Miíase/cirurgia , Doenças Orbitárias/cirurgia , Idoso , Animais , Dípteros , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Miíase/diagnóstico , Doenças Orbitárias/diagnóstico por imagemRESUMO
This study was to identify anti-metastatic active fractions and compounds of Bolboschoenus yagara (B. yagara). The results indicated that 50 µg/mL ethyl acetate fraction (Et) can dramatically inhibit mouse melanoma B16 cells migration and invasion in vitro. In B16 cells pulmonary and hepatic metastasis assays, 50 µg/mL Et alleviated mouse lung and liver weights, the number of metastatic nodules and the levels of TNF-α and IL-6 in mouse serum and organs. Histological studies showed that Et fraction was able to prevent liver and lung metastasis. And the inhibition of 50 µg/mL Et fraction against hepatic metastasis was almost equivalent to that of 1 µM TAK242. In addition, fourteen compounds of Et were quantified by HPLC analysis, in which, isocoumarins, stilbenes and xanthones obviously abated LPS-modulated B16 cells migration and invasion.[Formula: see text].
Assuntos
Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/farmacologia , Cyperaceae/química , Melanoma Experimental/tratamento farmacológico , Melanoma Experimental/patologia , Animais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Feminino , Interleucina-6/sangue , Interleucina-6/metabolismo , Lipopolissacarídeos/toxicidade , Neoplasias Hepáticas Experimentais/secundário , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/secundário , Camundongos Endogâmicos BALB C , Tubérculos/química , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: Macrophage accumulation in the vascular wall is a hallmark of atherosclerosis. Studies showed that shifting of oxidized lipids-induced inflammatory macrophages towards an anti-inflammatory phenotype by promoting oxidative metabolism attenuated atherosclerosis progression. Therefore, this study aimed to investigate whether metformin, which has ameliorated atherosclerosis in animal models and clinical trials, modulated oxidized low-density lipoprotein (Ox-LDL) induced inflammatory status in macrophages by regulating cellular oxidative metabolism. METHODS: Murine raw264.7 macrophages were incubated with Ox-LDL (50âµg/mL) in the presence or absence of metformin (15âµmol/L) for 24âh. Real-time polymerase chain reaction was used to quantify the transcription of classically activated (M1) pro-inflammatory and alternatively activated (M2) anti-inflammatory markers and mitochondrial DNA copy numbers. Cellular reactive oxygen species (ROS) production and mitochondrial membrane potential were detected by immunofluorescence. Cellular adenosine triphosphate (ATP) synthesis, glucose uptake, and lactic acid production were measured by commercial kit and normalized to cellular lysates. Western blotting analysis was performed to detect the expression of mitochondrial fusion/fission related proteins, enzymes mediating lipid metabolism and signaling pathway of glucose transport. Differences between groups were analyzed using one-way analysis of variance. RESULTS: Metformin improved Ox-LDL-impaired anti-inflammatory phenotype in raw264.7 macrophages as shown by up-regulated transcription of anti-inflammatory markers including interleukin 10 (0.76â±â0.04 vs. 0.94â±â0.01, Pâ=â0.003) and Resistin-like molecule alpha (0.67â±â0.08 vs. 1.78â±â0.34, Pâ=â0.030). Conversely, Ox-LDL-diminished phosphorylation of Akt was up-regulated by metformin treatment (0.47â±â0.05 vs. 1.02â±â0.08, Pâ=â0.040), associated with an improvement of mitochondrial function, characterized by decreased ROS generation (2.50â±â0.07 vs. 2.15â±â0.04, Pâ=â0.040), increased lipid oxidation, and elevated cellular ATP production (0.026â±â0.001 vs. 0.035â±â0.003, Pâ=â0.020). Moreover, metformin-mediated Akt activation increased Akt substrate of 160âkDa (AS160) phosphorylation (0.51â±â0.04 vs. 1.03â±â0.03, Pâ=â0.0041), promoted membrane translocation of glucose transporter 1, and increased glucose influx into the cells (4.78â±â0.04 vs. 5.47â±â0.01, Pâ<â0.001). CONCLUSION: This study suggested that targeting macrophage metabolism with new or existing drugs had therapeutic potential for the prevention and treatment of diabetes-accelerated atherosclerosis.
Assuntos
Lipoproteínas LDL/farmacologia , Metformina/farmacologia , Animais , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Células RAW 264.7 , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/efeitos dos fármacosRESUMO
Growing evidence indicates that the gut microbiota plays a significant role in the pathophysiological processes of obesity and its related metabolic symptoms in the host. Puerarin, an active ingredient in the root of Pueraria lobate has been suggested to have a potent anti-obesity effect. Herein, we tested whether this effect of puerarin is associated with changes in the gut microbiota. In addition to reducing body weight, inflammation, and insulin resistance, puerarin administration significantly altered the composition of the gut microbiota. Notably, puerarin treatment greatly increased the abundance of Akkermansia muciniphila, a mucin-degrading bacterium known to be beneficial for host metabolism and significantly downregulated in high-fat diet-fed mice. Further experiments revealed that puerarin increased intestinal expression levels of Muc2 and Reg3g and protected intestinal barrier function (normal permeability) by increasing the expression of ZO-1 and occludin in vivo and in vitro. These data suggest that puerarin's enriching effect on A. muciniphila is mediated, at least in part, by a host cellular response to protect the host from diet-induced metabolic disorders and other diseases.
Assuntos
Dieta Hiperlipídica/efeitos adversos , Microbioma Gastrointestinal , Infecções por Bactérias Gram-Negativas/microbiologia , Obesidade/etiologia , Verrucomicrobia , Akkermansia , Animais , Biomarcadores , Glicemia , Linhagem Celular Tumoral , Modelos Animais de Doenças , Regulação da Expressão Gênica , Humanos , Masculino , Camundongos , Obesidade/metabolismo , FenótipoRESUMO
In this study, we try to explore the effect of orbital decompression treatment on severe dysthyroid optic neuropathy.We retrospectively collected demographic and clinical characteristics of thyroid eye disease patients who performed orbital decompression. Then we analyzed the change of best-corrected visual acuity and exophthalmometry after surgery and the correlations among clinical parameters.A total of 22 cases (30 eyes) were included in the study. After orbital decompression, visual acuities improved in 16 eyes, declined in 8 eyes, and had no change in 5 eyes. Best-corrected visual acuity was significantly improved (0.1 vs 0.4, Pâ=â.039) and exophthalmometry was significantly declined (22.0âmm vs 16.5âmm, Pâ=â.001) after orbital decompression. Better postoperative best-corrected visual acuity was significantly correlated with better preoperative best-corrected visual acuity (râ=â0.718, Pâ<â.05), and with normal optic disc (râ=â0.568, Pâ<â.05), but not with age, exophthalmometry, keratopathy, and clinical activity score.These results showed that orbital decompression is a useful approach to manage dysthyroid optic neuropathy. The optimal time for surgery should be chosen based on clinical parameters, such as visual acuity and degree of crowding of orbital apex.
Assuntos
Descompressão Cirúrgica/métodos , Oftalmopatia de Graves/complicações , Síndromes de Compressão Nervosa/cirurgia , Doenças do Nervo Óptico/cirurgia , Órbita/cirurgia , Adulto , Feminino , Oftalmopatia de Graves/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade , Síndromes de Compressão Nervosa/etiologia , Doenças do Nervo Óptico/etiologia , Estudos Retrospectivos , Resultado do Tratamento , Acuidade VisualRESUMO
AIMS: The accumulation of lipids in macrophages contributes to the development of atherosclerosis. Cholesterol efflux of lipid-loaded macrophages mediated by ATP binding cassette (ABC) cholesterol transporters, on the other hand, has been shown to attenuate atherosclerosis progression in patients with unknown mechanism. We therefore sought to test the effect of metformin that reduced cardiovascular risk in diabetic patients independent of its hypoglycemia effect on cholesterol transport in murine raw264.7 macrophages. MATERIALS AND METHODS: Mouse raw264.7 macrophages were loaded with Ox-LDL (50⯵g/ml) for 24â¯h before incubated with metformin (15⯵M) for 24â¯h. Foam cell formation was assessed by Oil red staining and BIODIPY fluorescent staining as well as cholesterol-ester quantification by commercial kit. Cholesterol uptake and expression of scavenger receptors were detected by flow-cytometry. Cholesterol efflux capacity was measured by fluorescent plate-reader and ABC transporters were detected by Western Blots. Cytokines were detected by ELISA in supernatants and normalized by cellular lysates. KEY FINDINGS: Our results showed that metformin decreased oxidized low-density lipoprotein (Ox-LDL)-induced cholesterol accumulation and foam cell formation by increasing cholesterol efflux to HDL, which was associated with an upregulation of ABC transporter ABCG-1. Moreover, metformin increased Ox-LDL-impaired IL-10 secretion, an important anti-foam cell cytokine in atherosclerosis. SIGNIFICANCE: Our data highlighted the therapeutic potential of targeting macrophage cholesterol efflux with new or existing drugs for the possible reduction of foam cell formation in the prevention and treatment of diabetes-accelerated atherosclerosis.
Assuntos
Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Aterosclerose/prevenção & controle , Hipoglicemiantes/farmacologia , Lipoproteínas LDL/administração & dosagem , Metformina/farmacologia , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Animais , Colesterol/metabolismo , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Células Espumosas/efeitos dos fármacos , Células Espumosas/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Células RAW 264.7 , Regulação para Cima/efeitos dos fármacosRESUMO
BACKGROUND: S1PR1-STAT3 inter-regulatory loop was initially suggested to be oncogenic in several cancer cells. However, the clinical relevance of this mechanism in tumor progression, disease prognosis and drug response was not established. METHODS: The correlations between S1PR1 transcription, overall survival and chemotherapy response of GC patients were tested using a large clinical database. The relevance of S1PR1 expression and STAT3 activation in both tumor tissues and cancer cell lines was also tested. The effect of S1PR1 high expression achieved by persistent STAT3 activation on tumor cell drug resistance was investigated in vitro and in vivo. FINDINGS: An enhanced S1PR1 expression was highly related with a reduced overall survival time and a worse response to chemotherapy drug and closer correlation to STAT3 in gastric cancer patients. The issue chip analysis showed that the expressions of S1PR1 and STAT3 activation were increased in higher graded gastric cancer (GC) tissues. Cellular studies supported the notion that the high S1PR1 expression was responsible for drug resistance in GC cells through a molecular pattern derived by constitutive activation of STAT3. The disruption of S1PR1-STAT3 signaling significantly re-sensitized drug resistance in GC cells in vitro and in vivo. INTERPRETATION: S1PR1-STAT3 signaling may participate drug resistance in GC, thus could serve as a drug target to increase the efficacy of GC treatment. FUND: This work was supported by the National Natural Science Foundation of China (No. 81570775, 81471095), the grant from the research projects in traditional Chinese medicine industry of China (No. 201507004-2).
Assuntos
Resistencia a Medicamentos Antineoplásicos , Proteínas de Neoplasias/biossíntese , Receptores de Lisoesfingolipídeo/biossíntese , Fator de Transcrição STAT3/biossíntese , Transdução de Sinais , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/mortalidade , Animais , Antineoplásicos/administração & dosagem , Linhagem Celular Tumoral , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas de Neoplasias/genética , Receptores de Lisoesfingolipídeo/genética , Fator de Transcrição STAT3/genética , Receptores de Esfingosina-1-Fosfato , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/genética , Taxa de SobrevidaRESUMO
BACKGROUND: The polycystic ovary syndrome (PCOS) is a common metabolic and endocrine disorder with pathological mechanisms remain unclear. The following study investigates the ovarian hyperfibrosis forming via transforming growth factor-ß (TGF-ß) signaling pathway in Dehydroepiandrosterone (DHEA)- induced polycystic ovary syndrome (PCOS) rat model. We furthermore explored whether TGF-ßRI inhibitor (SB431542) decreases ovarian fibrosis by counterbalancing the expression of fibrotic biomarkers. METHODS: Thirty female Sprague-Dawley rats were randomly divided into Blank group (n = 6), Oil group (n = 6), and Oil + DHEA-induced model group (n = 6 + 12). The model groups were established by subcutaneous injection of DHEA for 35 consecutive days. The 12 successful model rats were additionally divided in vehicle group (n = 6) and SB431542-treated group (n = 6). Vehicle group and SB431542-treated group, served as administration group and were intraperitoneally injected with DMSO and SB431542 for additional 14 consecutive days. Ovarian morphology, fibrin and collagen localization and expression in ovaries were detected using H&E staining, immunohistochemistry and Sirius red staining. The ovarian protein and RNA were examined using Western blot and RT-PCR. RESULTS: In DHEA-induced ovary in rat, fibrin and collagen had significantly higher levels, while the main fibrosis markers (TGF-ß, CTGF, fibronectin, a-SMA) were obviously upregulated. SB431542 significantly reduced the expression of pro-fibrotic molecules (TGF-ß, Smad3, Smad2, a-SMA) and increased anti-fibrotic factor MMP2. CONCLUSION: TGF-ßRI inhibitor (SB431542) inhibits the downstream signaling molecules of TGF-ß and upregulates MMP2, which in turn prevent collagen deposition. Moreover, ovarian hyperfibrosis in DHEA-induced PCOS rat model could be improved by TGF-ßRI inhibitor (SB431542) restraining the transcription of accelerating fibrosis genes and modulating EMT mediator.
Assuntos
Desidroepiandrosterona/efeitos adversos , Síndrome do Ovário Policístico/etiologia , Síndrome do Ovário Policístico/metabolismo , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo , Animais , Biomarcadores , Modelos Animais de Doenças , Ciclo Estral , Feminino , Fibrose , Citometria de Fluxo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Hormônios/sangue , Imuno-Histoquímica , Síndrome do Ovário Policístico/patologia , RatosRESUMO
BACKGROUND: Cancer related inflammation plays a fatal role in the metastatic process, which can foster tumor growth, angiogenesis and dissemination. Sparstolonin B (SsnB), derived from Chinese medicine of the tubers of Scirpus yagara, is a TLR2 and TLR4 antagonists. It has exhibited multiple activities of anti-inflammatory, anti-cancer, anti-obesity and anti-hepatitis. However, whether SsnB is involved in the regulation of inflammation-induced tumor metastasis is not well elucidated. PURPOSE: The aim of this study was to investigate the effectiveness of SsnB as a treatment of inflammation-induced tumor metastasis and identify the underlying mechanisms of its anti-tumor metastatic activity. METHOD: The anti-tumor metastatic activity in vitro was estimated by MTT, wound-healing assay, matrigel invasion analysis and extracellular matrix adhesion assay. Mice lung metastasis and hepatic metastasis experiments were performed to assess the activities in vivo. Lungs or livers were weighed and the number of metastatic nodules was determined after mice were sacrificed. The levels of pro-inflammatory cytokines in the serum, lungs and livers were detected by using enzyme-linked immunosorbent assay (ELISA). Micro-metastasis nodules in lungs or livers were analyzed by histological examination. Immunohistochemistry and western blot analysis were conducted to determine protein expression. RESULT: Herein, SsnB dose-dependently inhibited cell migration and invasion in mouse melanoma B16 cells with or without stimulation of lipopolysaccharide (LPS), Pam3csk4 or molecules from damaged tumor cells (DTC-Ms). The expression of matrix metalloproteinases (MMP)-2 was also significantly abated by SsnB in LPS-modulated B16 cells. And SsnB reduced LPS-activated B16 cells adhesion to extracellular matrix components collagen I and fibronectin in a dose-dependent manner. In vivo, SsnB obviously attenuated LPS-activated pulmonary metastasis in mice by reduction the number of metastatic nodules on the lung surfaces, lung weight and levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serums and lungs. Moreover, in experimental hepatic metastasis model mice, SsnB remarkably repressed LPS-stimulated the number of metastatic nodules along with liver weight; and SsnB significantly suppressed LPS-activated increase levels of TNF-α and IL-6 in livers. Immunohistochemistry analysis indicated that SsnB inhibited the expression of TLR4 in livers. Furthermore, SsnB remarkably blocked p38 and ERK1/2 signaling pathway in LPS-induced B16 cells. P38 and ERK1/2 signaling silencing, using BIRB0796 (small molecular inhibitor of p38 MAPK) and PD184352 (inhibitor of MEK1/2 kinases that activate ERK1/2), significantly abated LPS-induced migration and invasion of B16 cells. CONCLUSION: The present study reports a novel use of SsnB in mitigating TLRs ligands-induced melanoma metastasis by inhibition of p38 and ERK1/2 pathway.
Assuntos
Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Inflamação/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Melanoma/tratamento farmacológico , Metástase Neoplásica/tratamento farmacológico , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Feminino , Inflamação/tratamento farmacológico , Interleucina-6/metabolismo , Lipopolissacarídeos/farmacologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Melanoma/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
This study performed an input-output structural decomposition analysis on changes in COD, ammonia nitrogen, SO2, NOx, soot and dust, industrial solid waste, and CO2 emission multipliers for 41 final products over the period 2007-2012 in China. The results show that during the examined period, emission multipliers were, in general, decreasing. The main driver of this was technical effects. The effects that made a significant contribution were concentrated in eight sectors: coal mining and washing; metals mining and quarrying; food and tobacco products; paper printing manufacturing; the chemical industry; non-metallic mineral products; metal smelting and rolling processing; and electricity, heat production, and supply. Moreover, the technical effects presented an obvious spillover. Although the contribution of the structural effects was far less than the technical ones, there were still some structural adjustments that led to significant synergistic mitigation. For example, the decrease in the direct demand of the agriculture products, electricity, and heat for food and tobacco products commonly reduced SO2, NOx, and CO2. In addition, four technical effects and most of the structural effects with high efficiency made small contributions. More than one third of the structural effects that showed obvious contributions played a positive role.
Assuntos
Carbono , Resíduos Industriais , China , Indústrias , Resíduos SólidosRESUMO
OBJECTIVE: To evaluate the potential for diagnosing colorectal cancer (CRC) from faecal metagenomes. DESIGN: We performed metagenome-wide association studies on faecal samples from 74 patients with CRC and 54 controls from China, and validated the results in 16 patients and 24 controls from Denmark. We further validated the biomarkers in two published cohorts from France and Austria. Finally, we employed targeted quantitative PCR (qPCR) assays to evaluate diagnostic potential of selected biomarkers in an independent Chinese cohort of 47 patients and 109 controls. RESULTS: Besides confirming known associations of Fusobacterium nucleatum and Peptostreptococcus stomatis with CRC, we found significant associations with several species, including Parvimonas micra and Solobacterium moorei. We identified 20 microbial gene markers that differentiated CRC and control microbiomes, and validated 4 markers in the Danish cohort. In the French and Austrian cohorts, these four genes distinguished CRC metagenomes from controls with areas under the receiver-operating curve (AUC) of 0.72 and 0.77, respectively. qPCR measurements of two of these genes accurately classified patients with CRC in the independent Chinese cohort with AUC=0.84 and OR of 23. These genes were enriched in early-stage (I-II) patient microbiomes, highlighting the potential for using faecal metagenomic biomarkers for early diagnosis of CRC. CONCLUSIONS: We present the first metagenomic profiling study of CRC faecal microbiomes to discover and validate microbial biomarkers in ethnically different cohorts, and to independently validate selected biomarkers using an affordable clinically relevant technology. Our study thus takes a step further towards affordable non-invasive early diagnostic biomarkers for CRC from faecal samples.
Assuntos
Biomarcadores Tumorais , Neoplasias Colorretais/diagnóstico , Disbiose/microbiologia , Fezes/microbiologia , Microbioma Gastrointestinal/genética , Idoso , Área Sob a Curva , Áustria , Estudos de Casos e Controles , China , Estudos de Coortes , Neoplasias Colorretais/complicações , Dinamarca , Disbiose/complicações , Feminino , Firmicutes/isolamento & purificação , França , Fusobacterium nucleatum/isolamento & purificação , Estudo de Associação Genômica Ampla , Humanos , Masculino , Metagenômica , Pessoa de Meia-Idade , Peptostreptococcus/isolamento & purificação , Curva ROCRESUMO
BACKGROUND: Neuroinflammation plays a key role in intracerebral hemorrhage (ICH)-induced secondary brain injury, but the specific roles of peripheral inflammatory cells such as macrophages and lymphocytes remain unknown. The purpose of this study was to explore the roles of macrophages, T lymphocytes, and the cytokines they secrete as potential targets for treating secondary brain injury after ICH. METHODS AND RESULTS: Our results showed that peripheral macrophages and T lymphocytes successively infiltrated the brain, with macrophage counts peaking 1 day after ICH and T-lymphocyte counts peaking after 4 days. These peaks in cellular infiltration corresponded to increases in interleukin (IL)-23 and IL-17 expression, respectively. We found that hemoglobin from the hematoma activated IL-23 secretion by infiltrating macrophages by inducing the formation of toll-like receptor (TLR) 2/4 heterodimer. This increased IL-23 expression stimulated γδT-cell production of IL-17, which increased brain edema and neurologic deficits in the model mice as a proinflammatory factor. Finally, we found that sparstolonin B (SsnB) could ameliorate brain edema and neurologic deficits in ICH model mice via inhibition of TLR2/TLR4 heterodimer formation, and notably, SsnB interacted with myeloid differentiation factor 88 Arg196. CONCLUSIONS: Together, our results reveal the importance of the IL-23/IL-17 inflammatory axis in secondary brain injury after ICH and thus provide a new therapeutic target for ICH treatment.