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1.
Acta Pharmaceutica Sinica B ; (6): 292-303, 2024.
Article in English | WPRIM | ID: wpr-1011234

ABSTRACT

This study examines inhibiting galectin 1 (Gal1) as a treatment option for hepatocellular carcinoma (HCC). Gal1 has immunosuppressive and cancer-promoting roles. Our data showed that Gal1 was highly expressed in human and mouse HCC. The levels of Gal1 positively correlated with the stages of human HCC and negatively with survival. The roles of Gal1 in HCC were studied using overexpression (OE) or silencing using Igals1 siRNA delivered by AAV9. Prior to HCC initiation induced by RAS and AKT mutations, lgals1-OE and silencing had opposite impacts on tumor load. The treatment effect of lgals1 siRNA was further demonstrated by intersecting HCC at different time points when the tumor load had already reached 9% or even 42% of the body weight. Comparing spatial transcriptomic profiles of Gal1 silenced and OE HCC, inhibiting matrix formation and recognition of foreign antigen in CD45+ cell-enriched areas located at tumor-margin likely contributed to the anti-HCC effects of Gal1 silencing. Within the tumors, silencing Gal1 inhibited translational initiation, elongation, and termination. Furthermore, Gal1 silencing increased immune cells as well as expanded cytotoxic T cells within the tumor, and the anti-HCC effect of lgals1 siRNA was CD8-dependent. Overall, Gal1 silencing has a promising potential for HCC treatment.

2.
Article in Chinese | WPRIM | ID: wpr-1019148

ABSTRACT

Objective To investigate the effect of galectin-1 preconditioning on pyroptosis of venti-lator-induced lung injury(VILI)in mice.Methods Thirty clean grade healthy male C57BL/6 mice,aged 6-8 weeks,weighing 22-30 g,were divided into three groups by random number table method:control group(group C),VILI group(group V),and galectin-1+VILI group(group G),10 mice in each group.After endotracheal intubation,group C kept spontaneous breathing for 4 hours,groups V and G kept me-chanical ventilation for 4 hours.One hour before endotracheal intubation,groups C and V were intraperito-neally injected with normal saline 0.75 ml,and group G was intraperitoneally injected with galectin-1 3 μg.Arterial blood was collected before endotracheal intubation and after spontaneous respiration or ventilation to detect PaO2.Then mice were sacrificed and bronchoalveolar lavage fluid(BALF)was collected.Concentra-tions of IL-1β and IL-18 in BALF were detected by ELISA.Lung tissue was collected for determination of the wet weight/dry weight ratio(W/D).The expression of GSDMD,caspase-1,and caspase-11 mRNA and protein in lung tissues were detected by qRT-PCR and Western blot.Pathological changes of the lungs were observed and scored by HE staining.Results Compared with group C,PaO2 were significantly decreased,W/D,concentrations of IL-1β and IL-18 in BALF,mRNA and protein expressions of GSDMD,caspase-1 and caspase-11,and lung injury score were significantly increased in groups V and G(P<0.05).Com-pared with group V,PaO2 was significantly increased,W/D,concentrations of IL-1β and IL-18 in BALF,mRNA and protein expressions of GSDMD,caspase-1,and caspase-11,and lung injury score were signifi-cantly decreased in group G(P<0.05).Conclusion Galectin-1 can increase PaO2 in mice and reduce IL-1β and IL-18 concentration,mRNA expression and protein content of classical non-classical pyroptosis pathway related genes,and reduce VILI in mice.

3.
Pesqui. vet. bras ; Pesqui. vet. bras;40(12): 1010-1017, Dec. 2020. tab, graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1155040

ABSTRACT

Canine melanoma is a frequently-occuring neoplasm in dogs and presents as malignant and highly metastatic in this context, studies that contribute to the understanding of the tumor microenvironment in melanoma include the role of galectins. Galectins are proteins of the family of animal lectins that display carbohydrate recognition domains. Galectin-1 and galectin-3 are associated with neoplastic transformation, neoplastic cell survival, angiogenesis, immune system evasion, and metastasis. The goal of this study was to establish a correlation between expression patterns of galectin-1 and galectin-3 and the different degrees of aggressiveness of canine melanoma, as well as to determine serum concentration of galectin-3 in dogs with melanoma. Galectin-1 and galectin-3 expression was analyzed by immunohistochemistry in 30 canine melanomas, six melanocytomas and nine metastatic lymph nodes from patients whose primary tumors were also processed and analyzed. Serum samples from 30 dogs were collected and galectin-3 concentration was determined by ELISA and compared to the samples of 10 healthy dogs. Canine melanoma samples expressed galectin-1 in the cytoplasm and presented a variable pattern of galectin-3 staining depending on melanoma aggressiveness. We observed a decrease in the percentage of cells with cytoplasmic galectin-3 immunolabeling simultaneous to the increased nuclear staining intensity, while there was also a decrease in the percent frequency of nuclear galectin-3 immunolabeled cells according to progression of melanoma, comparing the least to the most aggressive cases. Dogs with melanoma had increased serum levels of galectin-3 when compared to healthy animals, suggesting its potential biomarker of patients with melanoma.(AU)


O melanoma canino é uma neoplasia frequente em cães que apresenta um potencial maligno e metastático. Neste contexto, investigar o microambiente tumoral é fundamental para compreender os mecanismos intercelulares e intracelulares envolvidos no desenvolvimento e progressão da doença. Neste estudo, destacamos as galectinas, proteínas da família das lectinas animais que exibem domínios de reconhecimento à carboidratos; a galectina-1 e a galectina-3 estão associadas a transformação neoplásica, sobrevivência de células neoplásicas, angiogênese, evasão do sistema immune e desenvolvimento de metástases. O objetivo deste estudo foi determinar os padrões de expressão de galectina-1 e galectina-3 em diferentes graus de agressividade do melanoma canino, bem como dosar a concentração sérica de galectina-3 em cães com melanoma e comparar com cães saudáveis. A expressão de galectina-1 e galectina-3 foi analisada em 30 melanomas caninos, seis melanocitomas e nove linfonodos metastáticos. A galectina-3 sérica foi mensurada em 30 cães com melanoma e comparada a 10 cães saudáveis. No melanoma canino a expressão de galectina-1 foi citoplasmática e a expressão de galectina-3 foi variável de acordo com o grau de agressividade. Notou-se uma redução na porcentagem de células com imunomarcação de galectina-3 citoplasmática e um aumento simultâneo da intensidade de imunomarcação nuclear, enquanto houve também uma diminuição na frequência percentual de células com imunomarcação nuclear de acordo com a progressão do melanoma comparando-se os casos menos com os mais agressivos. Cães com melanoma apresentaram níveis séricos aumentados de galectina-3 quando comparados a animais saudáveis, mostrando seu uso potencial como biomarcador em pacientes com melanoma.(AU)


Subject(s)
Animals , Dogs , Immunohistochemistry , Galectin 1 , Galectin 3 , Dogs/abnormalities , Melanoma , Lectins
4.
Article in English | WPRIM | ID: wpr-766042

ABSTRACT

BACKGROUND: In this study, we investigate the expression of markers of angiogenesis and microvessel density (MVD) in cases of microcystic, elongated and fragmented (MELF) pattern, with its prognostic role in the survival of endometrioid endometrial adenocarcinomas (EA) patients. METHODS: In this study, 100 cases of EA, 49 cases with MELF pattern and 51 without, were immunohistochemically stained for galectin-1, vascular endothelial growth factor (VEGF), and MVD. Morphometry and statistical (univariate and multivariate) analyses were performed to assess overall survival (OS) and disease-free survival. RESULTS: The expression of VEGF (p<.001) and galectin-1 (p<.001), as well as MVD area (p<.001) and number of vessels/mm² (p<.050), were significantly higher in the +MELF pattern group compared to the –MELF group. A low negative correlation between MELF-pattern and the number of days of survival (p<.001, r=–0.47) was also found. A low positive correlation of MELF-pattern with galectin-1 expression (p<.001, r=0.39), area of vessels/mm² (p<.001, r=0.36), outcome of EA (p<.001, r=0.42) and VEGF expression (p<.001, r=0.39) suggests potential pathological relevance of these factors in the prognosis of EA. A univariate survival analysis indicated a role for all parameters of survival. Multivariate Cox proportional hazard regression analysis revealed that only area of vessels/mm² (hazard ratio [HR], 1.018; 95% confidence interval [CI], 1.002 to 1.033), galectin-1 (HR, 1.049; 95% CI, 1.025 to 1.074) and VEGF (HR, 1.049; 95% CI, 1.022 to 1.077) play key roles in OS. CONCLUSIONS: This study reports an increase in MVD, VEGF and galectin-1 expression in EA with MELF pattern and suggests that MELF pattern, along with the angiogenic profile, may be a prognostic factor in EA.


Subject(s)
Humans , Adenocarcinoma , Disease-Free Survival , Galectin 1 , Microvessels , Prognosis , Vascular Endothelial Growth Factor A
5.
Article in Chinese | WPRIM | ID: wpr-805377

ABSTRACT

Objective@#To observe the expression and clinical significance of galectin-1 in Chinese laryngeal squamous cell carcinoma patients.@*Methods@#Immunohistochemical technique was used to test the galecin-1 protein expression in the 50 laryngeal squamous cell carcinoma patients and Western blot method was used to test the galecin-1 protein expression in the 3 of the patients. SAS 9.4 software was used to analyze the relationship between the galectin-1 protein and the clinical degrees, pathological grades and lymph node metastasis in all the cases.@*Results@#In the 50 laryngeal squamous cell carcinoma patients, the positive rate of galectin-1 protein was 74% (37/50). The expression of galectin-1 protein was correlated with the severity of clinical staging, pathological differentiation and lymph node metastasis.@*Conclusions@#High expression of galectin protein related with the occurrence and development of head and neck squamous cell carcinomas.

6.
Article in Chinese | WPRIM | ID: wpr-515371

ABSTRACT

Objective To investigate the inhibitory effects and possible related mechanism of OTX008 [a selective inhibitor of galectin-1 (Galectin-1)] on retinal neovascularization (RNV) in mouse model of oxygeninduced retinopathy (OIR).Methods 7-day-old (P7) C57BL/6J mice were randomly (according to random number table) divided into 4 groups including normal group,OIR group,OIR-OTX008 group and OIRphosphate buffered saline (PBS) group.To establish the OIR mouse model,mice from all groups except normal group were expose to (75±2)% oxygen for 5 days and then to room air.OIR-OTX008 group received an intravitreal injection of 1 μl (0.25 μg/μl) OTX008 at P12,OIR-PBS group received the equal volume (1 μl) of PBS injection.Mice from 4 groups were euthanized at P17,and retinas were collected for molecular biological analysis and morphological study.RNV was evaluated by counting the number ofpre-retinal neovascular nuclei and the whole-mount immunofluorescent staining of mouse retina.Cyrosections of retinas were imaged via confocal microscopy to observe the enrichment of staining of Galectin-1.Protein levels of Galectin-1,Neuropilin-1 and phosphorylation of vascular endothelial growth factor receptor 2 (pVEGFR2) were determined with Western blot.Results At P17,Galectin-1 expressed higher in retinal ganglion cell layer,inner plexiform layer and inner nuclear layer from OIR group and OIR-PBS group than normal group.Galectin-1 expressed less in cryosection retinas from OIR-OTX008 group than OIR group and OIR-PBS group.The numbers ofpre-retinal neovascular cell nuclei from OIR group and OIR-pBS group were obviously more than that from normal group (t=9.314,P<0.05).The number of pre-retinal neovascular cell nuclei from OIR-OTX008 group were obviously lower than those from OIR group and OIR-PBS group (t=8.038,7.774;P<0.05).The RNV tufts area (t=13.250,12.570),non-perfusion area (t=15.590,12.430) and hypoxic area (t=9.542,9.928) from OIR-OTX008 group were significantly smaller than those in OIR group and OIR-PBS group (P< 0.05).Protein levels of Galectin-1 (t=24.800,23.060),Neuropilin-1 (t=4.120,3.530) and pVEGFR2 (t=25.880,15.480) in the OIR-OTX008 group were significantly down-regulated than those from OIR group and OIR-PBS group (P<0.05).Conclusion Intravitreal injection of OTX008 inhibits RNV and ameliorates retinal hypoxia in mice model of OIR possibly through down-regulating Galectin-1,Neurolinpin-1 and pVEGFR2.

7.
Int. j. morphol ; 34(2): 742-751, June 2016. ilus
Article in English | LILACS | ID: lil-787063

ABSTRACT

Diabetes mellitus (DM), is a metabolic disease occurring via insulin secretion deficiency from the pancreas and/or an insufficiency of tissue response to insulin. The present study is intended to show of immunolocalizations of beta-galactose-binding proteins Galectin-1 and Galectin-3 in diabetic rat ovarium and their relationship with diabetes. In this study, 8 to 10-week-old, 250­300 g weighing 50 mature female rats were used, in order to establish diabetes mellitus in those animals, 60 mg/kg intravenous streptozotocin was injected to each animal. After death, diabetics and non-diabetics rats's routine tissue processing steps is done to rat ovarial tissues for immunohistochemical investigation. Strong expressions of Galectin-1 and Galectin-3 were observed in the ovarial germinal epithelium and vascular endothelial. While the strong intense expression of Galectin-1 was seen in the zona pellucida, Galectin-3 expression was strongest in the cytoplesmic regions of cells. Zona pellucida has 3 protein complexes (ZP1, ZP2 and ZP3) in rats and in humans and they have the capability of recognizing the carbonhydrate fields in tissues. The strong expression of galectins in those regions could be the result of carbonhydrate binding properties expression of Gal-3 in the cytoplasmic regions of growing follicles could suggest the idea that Gal-3 could have effects on follicle growth. In conclusion, beta galactose-binding proteins Gal-1 and Gal-3 had stronger immunolocalization in diabetic rat ovarium when compared to the controls. Diabetes could increase the Gal-1 and Gal-3 expressions in the ovarial tissue.


La diabetes mellitus (DM) es una enfermedad metabólica debido a una deficiencia en la secreción de insulina por parte del páncreas o por una insuficiente respuesta de los tejidos a la insulina. El objetivo fue demostrar la inmunolocalización de las proteínas de unión beta-galactosa Galectina-1 y Galectina-3 en los ovarios de ratas diabéticas y su relación con la diabetes. Fueron utilizadas 50 ratas hembras maduras entre 8­10 semanas de edad, con un peso de 250­300 g. Con el fin de desarrollar DM en los animales, se inyectó a cada uno 60 mg/kg de estreptozotocina vía intravenosa. Después de la eutanasia, se realizó el procesamiento de rutina de los tejidos de las ratas diabéticas y no diabéticas para evaluar los tejidos ováricosa través de inmunohistoquímica. Se observaron expresiones fuertes de la Galectina-1 y Galectina-3 en el epitelio germinal y epitelio endotelial vascular del ovario. Si bien la fuerte e intensa expresión de Galectina-1 se observó en la zona pelúcida, la Galectina-3 tuvo una expresión más fuerte en las regiones de las células citoplasmáticas. La zona pelúcida tiene 3 complejos de proteínas (ZP1, ZP2 y ZP3) en ratas y en seres humanos y tienen la capacidad de reconocer los campos de carbohidratos en los tejidos. La fuerte expresión de las galectinas de esas regiones podría ser el resultado de las propiedades de unión a carbonhidratos expresión de Gal-3 en las regiones citoplasmáticas de los folículos en crecimiento, pudiendo sugerir que Gal-3 podría tener efectos sobre el crecimiento del folículo. En conclusión, las proteínas de unión beta-galactosa Gal-1 y Gal-3 tienen una mayor inmunolocalización en los ovarios de ratas diabéticas, en comparación a los controles. La diabetes podría incrementar las expresiones de Gal-1 y Gal-3 en el tejido ovárico.


Subject(s)
Animals , Female , Rats , Diabetes Mellitus/metabolism , Galectin 1/metabolism , Galectin 3/metabolism , Ovary/metabolism , Immunohistochemistry
8.
Medicina (B.Aires) ; Medicina (B.Aires);74(4): 321-325, ago. 2014. ilus
Article in Spanish | LILACS | ID: lil-734393

ABSTRACT

Al producirse una lesión de médula espinal (LME), un sinnúmero de proteínas inhibidoras de la regeneración axonal ocupan el sitio de lesión en forma secuencial. La primer proteína en llegar al mismo se conoce como semaforina 3A (Sema3A), siendo además una de las más potentes por su acción de inhibir la regeneración axonal. A nivel mecanístico la unión de esta proteína al complejo-receptor neuronal neuropilin-1 (NRP-1)/PlexinA4 evita que se produzca regeneración axonal. En este trabajo de revisión se discutirá la acción de galectin-1 (Gal-1), una proteína endógena de unión a glicanos, que selectivamente se une al complejo-receptor NRP-1/PlexinA4 de las neuronas lesionadas a través de un mecanismo dependiente de interacciones lectina-glicano, interrumpiendo la señalización generada por Sema3A y permitiendo de esta manera la regeneración axonal y recuperación locomotora luego de producirse la LME. Mientras ambas formas de Gal-1 (monomérica y dimérica) contribuyen a la inactivación de la microglia, solo la forma dimérica de Gal-1 es capaz de unirse al complejo-receptor NRP-1/PlexinA4 y promover regeneración axonal. Por lo tanto, Gal-1 dimérica produce recuperación de las lesiones espinales interfiriendo en la señalización de Sema3A a través de la unión al complejo-receptor NRP-1/PlexinA4, sugiriendo el uso de esta lectina en su forma dimérica para el tratamiento de pacientes con LME.


When spinal cord injury (SCI) occurs, a great number of inhibitors of axonal regeneration consecutively invade the injured site. The first protein to reach the lesion is known as semaphorin 3A (Sema3A), which serves as a powerful inhibitor of axonal regeneration. Mechanistically binding of Sem3A to the neuronal receptor complex neuropilin-1 (NRP-1) / PlexinA4 prevents axonal regeneration. In this special article we review the effects of galectin-1 (Gal-1), an endogenous glycan-binding protein, abundantly present at inflammation and injury sites. Notably, Gal1 adheres selectively to the NRP-1/PlexinA4 receptor complex in injured neurons through glycan-dependent mechanisms, interrupts the Sema3A pathway and contributes to axonal regeneration and locomotor recovery after SCI. While both the monomeric and dimeric forms of Gal-1 contribute to ’switch-off’ classically-activated microglia, only dimeric Gal-1 binds to the NRP-1/PlexinA4 receptor complex and promotes axonal regeneration. Thus, dimeric Gal-1 promotes functional recovery of spinal lesions by interfering with inhibitory signals triggered by Sema3A adhering to the NRP-1/PlexinA4 complex, supporting the use of dimeric Gal-1 for the treatment of SCI patients.


Subject(s)
Animals , Humans , Mice , Axons/physiology , Galectin 1/physiology , Nerve Regeneration/physiology , Spinal Cord Injuries/physiopathology , Microglia/metabolism , Nerve Tissue Proteins/metabolism , Neuropilin-1/metabolism , Receptors, Cell Surface/metabolism , /physiology
9.
Korean Circulation Journal ; : 223-230, 2013.
Article in English | WPRIM | ID: wpr-209911

ABSTRACT

BACKGROUND AND OBJECTIVES: Existing data on the spatiotemporal expression patterns of a variety of galectins in murine atherosclerosis are limited. We investigated the expression levels of galectins, and their in vivo spatiotemporal expression patterns and statin responsiveness in the inflamed atherosclerotic plaques of apolipoprotein E (apoE)-/- mice. MATERIALS AND METHODS: Galectins expression patterns in aortic atherosclerotic plaques and serum galectin-3 levels were investigated in 26-week-old apoE-/- (n=6) and C57BL/6 mice (n=9). To investigate the spatial and temporal patterns of galectin-1 and galectin-3 in plaques, high-cholesterol diet-fed 26-week-old (n=12) and 36-week-old apoE-/- mice (n=6) were sacrificed and their aortas were examined for galectins' expression using immunoblot analysis and immunohistochemical stain. 36-week-old apoE-/- mice were treated with atorvastatin (n=3, 0.57 mg/kg/day) for the evaluation of its effect on aortic galectins' expression. RESULTS: Immunoblot analyses showed that galectin-1 and galectin-3 were the predominant galectins expressed in murine atherosclerosis. The serum galectin-3 level was significantly higher in apoE-/- mice (p<0.001). While galectin-1 was weakly expressed in both intimal plaques and the media of atherosclerotic aortas, galectin-3 was heavily and exclusively accumulated in intimal plaques. Galectin-3 distribution was colocalized with plaque macrophages' distribution (r=0.66). As the degree of plaque extent and inflammation increased, the intraplaque galectin-3 expression levels proportionally elevated (p<0.01 vs. baseline), whereas galectin-1 expression had not elevated (p=0.14 vs. baseline). Atorvastatin treatment markedly reduced intraplaque galectin-3 and macrophage signals (p<0.001 vs. baseline), whereas it failed to reduce galectin-1 expression in the aortas. CONCLUSION: Galectin-3 is the predominant gal and is colocalized with macrophages within atherosclerotic plaques. Intraplaque galectin-3 expression reflects the degree of plaque inflammation.


Subject(s)
Animals , Mice , Aorta , Apolipoproteins , Atherosclerosis , Galectin 1 , Galectin 3 , Galectins , Heptanoic Acids , Inflammation , Macrophages , Plaque, Atherosclerotic , Pyrroles , Atorvastatin
10.
Journal of Clinical Pediatrics ; (12): 949-952, 2013.
Article in Chinese | WPRIM | ID: wpr-441201

ABSTRACT

Objectives To investigate the dynamic change of galectin-1 in myocardium of mice with viral myocarditis (VMC). Methods A total of 58 male BALB/c mice were selected and randomly divided into CVB3 group (n=50) and control group (n=8). Mice in CVB3 group were infected with 0.10 ml 10-5/L CVB3 through intraperitoneal inoculation, and 8 mice were killed on day 7, 10, 14 and 28 respectively after inoculation. Mice in control group inoculated with 0.10 ml eagle reagent was killed on day 28. The myocardial pathological changes were observed using light microscope. In addition, expressions of serum galectin-1 were detected by ELISA and expressions of myocardial galectin-1 were detected by real-time quantitative-polymerase chain reaction (RQ-PCR). Results Compared with control group, the myocardial expression of galectin-1 mRNA in CVB3 group was obviously increased on day 7 and day 10 (all P<0.05);the serum concentration of galectin-1 in CVB3 group was obviously increased on day 7, day 10 and day 14 (all P<0.05). Conclusion The expressions of galectin-1 in myocardium of mice with viral myocarditis were in dynamic changes. Galectin-1 may play a role in the pathogenesis and development of viral myocarditis.

11.
Rio de Janeiro; s.n; 2012. 116 f p.
Thesis in Portuguese | LILACS | ID: lil-756637

ABSTRACT

O Aspergillus fumigatus é o principal agente etiológico da aspergilose invasiva, uma infecção fúngica oportunista que acomete, principalmente, pacientes de Unidades Hematológicas, como aqueles com neutropenia profunda e prolongada. Após a filamentação este fungo angioinvasivo é capaz de ativar e causar danos em células endoteliais de veia umbilical humana (HUVEC) que passam a expressar um fenótipo pró-trombótico. A ativação destas células, dependente de contato célulacélula, é mediada por TNF-α e caracterizada pela expressão de moléculas próinflamatórias, como citocinas, quimiocinas e moléculas de adesão. Recentemente, nosso grupo comparou a ativação endotelial de HUVECs desafiadas com cepas selvagens e uma cepa mutante para o gene UGM1. Nestes experimentos a cepa mutante Δugm1, que apresenta um fenótipo de maior produção de galactosaminogalactana (GAG) na parede celular, mostrou um fenótipo hiperadesivo e uma capacidade maior de ativar células endoteliais. Entretanto, os receptores e as vias de sinalização envolvidos nesta ativação permanecem desconhecidos. Assim, o objetivo deste trabalho foi verificar as proteínas envolvidas nestes processos através do estudo das proteínas diferencialmente expressas nas HUVECs após a interação com A. fumigatus, usando a técnica proteômica 2D-DIGE. Brevemente, as HUVECs foram infectadas com tubos germinativos da cepa selvagem (AF293) e da cepa Δugm1 de A. fumigatus. Em seguida, as proteínas foram marcadas com diferentes fluorocromos e separadas por eletroforese bidimensional. A análise quantitativa foi realizada utilizando o software DeCyder...


Aspergillus fumigatus is the main etiological agent of invasive aspergillosis, the main opportunistic fungal infection of Hematologial Unity’s patients, especially those with long-term neutropenia. Upon filamentation, this angioinvasive fungus can activate and damage the human umbilical vein endothelial cells (HUVEC), which in response switch to a pro-thrombotic phenotype. HUVEC activation is mediated by TNF-α once cell-cell contact occurs. This activation is characterized by the expression of pro-inflammatory molecules such cytokines, chemokines and adhesion molecules. Recently, our group performed the comparison of HUVEC activation upon interaction with a wild type and the UGM1 mutant strains of A. fumigatus. The Δugm1 strain, which presents an increased production of the cell wall galactosaminogalactan, showed a hyper adherent phenotype and an increased capability to cause endothelial cell stimulation and activation, when compared with the wild type strain. The receptors involved in the pathogen-host interaction or the signaling pathways after endothelial activation by A. fumigatus remain unknown. Thus, the aim of this study was to investigate the differentially expressed proteins in HUVECs upon interaction with A. fumigatus, using the 2D-DIGE proteomic approach. Briefly, HUVECs were challenged with germlings of A. fumigatus wild type Af293 and Δugm1 strains and then submitted to protein extraction. The total HUVEC protein extracts were labeled with different CyDyes and fractionated by 2D electrophoresis. Quantitative analysis to determine the differences in protein abundance amongst interacted cells vs. control endothelial cells was performed using the software DeCyder. Five differentially expressed proteins were identified by MS/MS including galectin-1 and annexin A2, both overexpressed after the interaction. These two proteins are described elsewhere to be associated with host-pathogen interaction...


Subject(s)
Aspergillus fumigatus , Human Umbilical Vein Endothelial Cells , Proteome , Endothelial Cells , Galectin 1 , Genome , Fungal Proteins/analysis
12.
Braz. j. otorhinolaryngol. (Impr.) ; Braz. j. otorhinolaryngol. (Impr.);76(2): 213-218, mar.-abr. 2010. ilus, tab
Article in English, Portuguese | LILACS | ID: lil-548323

ABSTRACT

A fisiopatologia da polipose rinossinusal não é totalmente compreendida, apesar de várias hipóteses em relação ao seu processo inflamatório. OBJETIVOS: Estudo prospectivo da expressão dos genes das proteínas, anexina-1 e a galectina-1, que têm ação anti-inflamatória, e sua modulação pelo glicocorticoide. MATERIAL E MÉTODOS: Onze pacientes portadores de polipose rinossinusal tiveram biopsiados seus pólipos em dois momentos: na ausência de glicocorticoide sistêmico, e na sua presença. Nas duas amostras, foi avaliada a expressão desses genes e comparada com a expressão na mucosa nasal normal do meato médio. RESULTADOS: Verificou-se que a média de expressão dos genes que codifica a anexina-1 e galectina-1 estava predominantemente aumentada, independente do uso do glicocorticoide em relação à mucosa nasal controle. Entretanto, nos pólipos sem uso de corticoide, a média de expressão do gene da anexina-1 foi significativamente maior do que nos pólipos que estavam sob uso de glicocorticoide. Com relação à galectina-1 não houve diferença significativa entre as médias de expressão antes e após o uso de glicocorticoide sistêmico. CONCLUSÃO: Os genes apresentaram um aumento da expressão na mucosa nasal polipoide, independente do uso do glicocorticoide, porém a relação destes dois genes das proteínas anti-inflamatórias com o glicocorticoide não ocorreu da mesma maneira.


Rhinosinusal polyps physiopathology is not fully understand, despite numerous hypotheses regarding its inflammatory process. AIMS: a prospective study regarding the gene expression of proteins: anexin-1 and galectin-1, which has an anti-inflammatory action and is modulated by steroids. MATERIALS AND METHODS: eleven patients with rhinosinusal polyps suffered a biopsy of their polyps at two moments: in the absence of systemic steroids and during its use. In the two samples we assessed the expression of these genes and compared it to the normal nasal mucosa in the middle meatus. RESULTS: We noticed that the mean expression of the genes which code anexin-1 and galectin-1 was predominantly increased, regardless of the use of steroids in relation to the control nasal mucosa. Notwithstanding, in polyps without the use of steroids, the mean gene expression of anexin-1 was significantly higher than in the polyps which were under the use of steroids. Regarding galectin-1, there was no significant difference between the expression mean values before and after the use of systemic steroids. CONCLUSION: The genes present an expression increase in the polyp mucosa, regardless of the use of steroids; nonetheless, the relationship of these two genes of anti-inflammatory proteins with steroids did not happen the same way.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Annexin A1/genetics , Anti-Inflammatory Agents/therapeutic use , Betamethasone/therapeutic use , Galectin 1/genetics , Glucocorticoids/therapeutic use , Nasal Polyps/drug therapy , Annexin A1/metabolism , Case-Control Studies , Galectin 1/metabolism , Gene Expression Regulation/drug effects , Nasal Polyps/metabolism , Prospective Studies
13.
Article in Korean | WPRIM | ID: wpr-19735

ABSTRACT

BACKGROUND: Galectin-1 (Gal-1) is a member of the galectin family of proteins, which are carbohydrate-binding proteins with an affinity for beta-galactosides. Gal-1 is differentially expressed by various normal and pathological tissues and it performs polyvalent, wide-ranging biological activities. A Gal-1 expression or over-expression in tumors and/or in the tissue surrounding them must be considered as a sign of malignant tumor progression that is often related to tumor metastasis. Although Gal-1 also plays important roles for tumorigenesis and tumor progression, the expression of Gal-1 in melanocytic nevus, dysplastic nevus and malgant melanoma has not yet been investigated. OBJECTIVE: We wanted to investigate and compare the expression of Gal-1 in melanocytic nevus, dysplastic nevusand malignant melanoma. METHODS: The paraffin-embedded specimens of 9 cases of malignant melanoma (MM), 6 cases of dysplastic nevus (DN) and 6 cases of intradermal nevus (IN) were subjected to immunohistochemical staining for Gal-1. RESULTS: The percentage of positive cells for Gal-1 in the MM was significantly higher than that of the DN and IN (p<0.01). The staining intensity of the positive cells for Gal-1 was the highest also in the MM. Meanwhile Gal-1 was more strongly expressed in highly atypical (more pleomorphic, more atypical mitoses) areas of the melanoma tissues. But there was no significant difference between the DN and IN for the expression of Gal-1. LIMITATION: This study is restricted to a small number of patients. CONCLUSION: The present study suggests that Gal-1 is more strongly expressed in malignant melanoma than in melanocytic nevus and dysplastic nevus. Interestingly, Gal-1 was more strongly expressed in the highly atypical portions of the melanoma tissue. Gal-1 might well contribute to the tumorigenesis and malignancy of melanocytes.


Subject(s)
Humans , Benzamides , Cell Transformation, Neoplastic , Dysplastic Nevus Syndrome , Galectin 1 , Galectins , Melanoma , Neoplasm Metastasis , Nevus, Intradermal , Nevus, Pigmented , Proteins , Tyrosine
14.
Article in English | WPRIM | ID: wpr-58677

ABSTRACT

PURPOSE: Radiation-induced pulmonary fibrosis (RIF) is a significant complication of radiotherapy for lung cancer. Despite the large number of studies, the molecular mechanisms of RIF are poorly understood. Therefore, the complex protein expression pattern in RIF was characterized by identifying the proteins with an altered expression level after thorax irradiation using two-dimensional electrophoresis (2-DE) and mass spectrometry. MATERIALS AND METHODS: A mouse model of RIF was used to examine the alteration of the lung proteome because of availability of murine data related to human cases and the abundance of murine fibrotic lung samples. A mouse model of RIF was induced in radiosensitive C57BL/6 mice. Twenty-one weeks after 25 Gy irradiation, hematoxylin-eosin staining and hydroxyproline assay confirmed the early-phase pulmonary fibrosis. RESULTS: Lung samples from the irradiated and age-matched control mice were used to generate 16 high quality 2-DE gels containing approximately 1,000 spots. Of the 31 significantly up- or down-regulated protein spots, 17 were identified by MALDI-TOF/MS. CONCLUSIONS: Two important upregulated proteins were found, the alpha-protease inhibitor and galectin-1, which might be used as potential markers for the early phase of RIF.


Subject(s)
Animals , Humans , Mice , Electrophoresis , Galectin 1 , Gels , Hydroxyproline , Lung , Lung Neoplasms , Mass Spectrometry , Proteome , Proteomics , Pulmonary Fibrosis , Radiotherapy , Thorax
15.
Article in Chinese | WPRIM | ID: wpr-532153

ABSTRACT

Objective To observe the changes of LoVo cell growth by galectin-1.Methods Eukaryotic expression vector of galectin-1 was constructed and transfected into LoVo cells using lipofectamineRM 2000.Immunochemistry was employed to detect galectin-1 expresson.LoVo cell proliferation and apoptosis were observed.Results Galectin-1 eukaryotic expression vector pEGFP-C1/GAL1 was successfully constructed.Three cell clones,p-GAL1-LoVo and p-LoVo,transfected with pEGFP-C1/GAL1 and pEGFP-C1 correspondingly,and LoVo were cultured successfully.Galectin-1 expression downregulated Bcl-2 level only occurring in p-GAL1-LoVo cells.p-GAL1-LoVo cell proliferation was similar to p-LoVo and LoVo cells,but p-GAL1-LoVo cell apoptosis was increased(9.61?0.56)%,as compared with p-LoVo and LoVo cells,[(3.56?0.53)% and(3.46?0.46)% respectively](P

16.
Article in Chinese | WPRIM | ID: wpr-558326

ABSTRACT

Objective To observe the influence on adhesion between LST-R1 cell and collagen Ⅰ mediated by galectin-1 antibody inhibition. Methods Laser confocal scanning microscopy was employed to detect the expression of galectin-1 on LST-R1 cell membrane.LST-R1 cells inhibited by galectin-1 antibody or not were seeded in the 48-well plate coated with collagen Ⅰ (20?g/well), and the morphology of the adhesive LST-R1 cells was observed and the adhesive cells were counted. Results Laser confocal scanning microscopy showed the expression of galectin-1 on the membrane of LST-R1 cells.The binding rate between galectin-1 and its antibody was 38.2%?0.92%. In the galectin-1 antibody inhibiting group,more and more irregular angular adhesive cells appeared, and most of the adhesive cells were growing in singles, while the adhesive cells,mainly clustered, in the control group were round or roudish. The numbers of adhesive LST-R1 cells in the antibody inhibiting group and the control group were 40 4 and 30 4 respectively (P

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