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1.
Rev. Assoc. Med. Bras. (1992) ; 68(2): 147-151, Feb. 2022. tab
Article in English | LILACS | ID: biblio-1365347

ABSTRACT

SUMMARY OBJECTIVE: Beta-thalassemia minor is a blood disease caused by a hereditary decrease in beta-globin synthesis, frequently leading to hypochromic microcytic anemia. Formerly called endothelial cell-specific molecule 1, endocan is a proteoglycan released by vascular endothelial cells in many organs. Our aim was to investigate the relationship between the beta-thalassemia minor patients and the healthy control group in terms of serum endocan level. METHODS: The study was performed in a total of 80 subjects. They were divided into two groups, the beta-thalassemia minor group (n=40) and the healthy control group (n=40). Serum endocan levels, age, sex, body mass index value, and tobacco use data of these groups were compared. RESULTS: No statistically significant difference was detected between the two groups in terms of age, sex, and body mass index values (p>0.05). Endocan levels were measured to be 206.85±88.1 pg/mL in the beta-thalassemia minor group and 236.1±162.8 pg/mL in the control group with no significant difference between the groups in terms of serum endocan levels (p>0.05). CONCLUSIONS: In our study, there was no change in endocan level in beta-thalassemia minor. This might be because serum endocan levels are affected by multi-factorial reasons. Serum endocan levels may be altered secondarily to decreased beta-globin chain, increased sympathetic activity due to anemia, or platelet dysfunction induced by oxidative stress in beta-thalassemia minor. Further multicenter studies involving more patients are necessary to demonstrate this.


Subject(s)
Humans , Proteoglycans , beta-Thalassemia , Neoplasm Proteins , Biomarkers , Body Mass Index , Endothelial Cells
2.
Arq. bras. cardiol ; 116(4): 756-762, abr. 2021. tab, graf
Article in English, Portuguese | LILACS | ID: biblio-1285206

ABSTRACT

Resumo Fundamento: Sugere-se que a serglicina tenha funções importantes na estabilização da fibrina e inflamação, mas há informações limitadas sobre seu valor clínico para a doença cardíaca aterosclerótica. Objetivo: O objetivo do presente estudo é descobrir os níveis séricos de serglicina em pacientes com infarto agudo do miocárdio e nos indivíduos do grupo controle; e investigar a associação entre os níveis de serglicina com marcadores de inflamação e marcadores de tamanho do infarto. Métodos: A população do estudo consistiu em 75 pacientes com infarto do miocárdio com supradesnivelamento do segmento ST (IAMCSST) e 57 pacientes com artérias coronárias normais (NCA) (grupo controle). As características dos pacientes, os níveis séricos de serglicina, os níveis de proteína C-reativa ultrassensível (PCR-us), os níveis máximos de troponina T e outros parâmetros bioquímicos foram registrados. O valor de p<0,05 foi considerado estatisticamente significativo. Resultados: O grupo controle consistiu em indivíduos mais jovens e que fumam menos do que os do grupo IAMCSST. O número de mulheres no grupo controle foi maior do que no grupo IAMCSST. Os níveis séricos de serglicina foram significativamente maiores no grupo IAMCSST do que no grupo controle (102,81±39,42 vs. 57,13±32,25, p<0,001). As análises de correlação revelaram uma correlação positiva significativa entre a serglicina e a troponina (correlação de postos de Spearman: 0,419; p<0,001) e entre a serglicina e a proteína C-reativa ultrassensível (correlação de postos de Spearman: 0,336; p<0,001). A análise de regressão logística multivariada demonstrou que os níveis séricos de serglicina apresentaram-se independentemente associados com IAMCSST. Usando um nível de corte de 80,47 μg/L, o nível de serglicina foi preditor da presença de IAMCSST com uma sensibilidade de 75,7% e especificidade de 68,4%. Conclusão: Os níveis séricos de serglicina apresentaram-se significativamente maiores no grupo IAMCSST do que no grupo controle. Os níveis de serglicina sérica mostraram-se positivamente correlacionados com os níveis de proteína C-reativa ultrassensível e troponina.


Abstract Background: It is suggested that serglycin has important functions in fibrin stabilization and inflammation but there is limited information on its clinical value for atherosclerotic heart disease. Objective: The purpose of this study is to find out serum serglycin levels in acute myocardial infarction patients and in the control group individuals; and to investigate the association between serglycin levels with inflammation markers and infarct size markers. Methods: The study population consisted of 75 patients with ST-segment elevation myocardial infarction (STEMI) and 57 patients with normal coronary arteries (NCA) (control group). Patient characteristics, serum serglycin levels, high-sensitivity C-reactive protein (hs-CRP) levels, peak troponin T levels and other biochemical parameters were recorded. A p value <0.05 was considered statistically significant. Results: The control group consisted of individuals who are younger and smoke less than those of the STEMI group. The number of females in the control group was higher than in the STEMI group. Serum serglycin levels were significantly higher in the STEMI group than in control group (102.81±39.42 vs. 57.13±32.25, p<0.001). Correlation analyses revealed a significant positive correlation between serglycin and troponin (Spearman's Rho: 0.419; p<0.001) and between serglycin and hs CRP (Spearman's Rho: 0.336; p<0.001). Multivariate logistic regression analysis demonstrated that serum serglycin levels were independently associated with STEMI. Using a cutoff level of 80,47 μg/L, the serglycin level predicted the presence of STEMI with a sensitivity of 75.7% and specificity of 68.4%. Conclusion: Serum serglycin levels were significantly higher in the STEMI group than in the control group. Serum serglycin levels were positively correlated with both hs CRP levels and troponin levels.


Subject(s)
Humans , Female , Percutaneous Coronary Intervention , ST Elevation Myocardial Infarction , Myocardial Infarction , Proteoglycans , Biomarkers , Vesicular Transport Proteins
4.
Braz. oral res. (Online) ; 35: e005, 2021. tab, graf
Article in English | LILACS, BBO | ID: biblio-1132742

ABSTRACT

Abstract: Endocan, a 50 kDa soluble proteoglycan, also called endothelial cell-specific molecule-1 (ESM-1), is involved in many major cellular activities and has been reported to be overexpressed in inflammatory conditions. This study aims to determine ESM-1 levels in gingival crevicular fluid (GCF) samples from individuals with periodontitis to determine the correlation between the levels of lymphocyte-function-associated antigen-1 (LFA-1), intercellular adhesion molecule-1 (ICAM-1), and clinical findings of periodontitis. This study enrolled 27 individuals diagnosed with Stage III-Grade C generalized periodontitis and 16 individuals as healthy controls. Bleeding on probing (BOP), probing pocket depth (PPD), and clinical attachment loss (CAL) were calculated. Enzyme-linked immunosorbent assay (ELISA) test was used for detecting the levels of ESM-1, ICAM-1, and LFA-1 in GCF samples. PPD, BOP, CAL, and GCF volumes were significantly increased in patients with periodontitis in comparison to the control group (p < 0.001). The total amount of ESM-1, ICAM-1, and LFA-1 levels in GCF were increased in the periodontitis group (p < 0.001). ESM-1 level correlated with PPD, BOP, and CAL (p < 0.05). ICAM-1 level correlated with BOP and CAL (p < 0.05). LFA-1 level correlated with PPD and CAL (p < 0.05). Our data indicate that ESM-1, ICAM-1, and LFA-1 levels are increased in GCF of patients with periodontitis. These molecules could be associated with the pathogenesis and progression of periodontal disease.


Subject(s)
Humans , Periodontitis , Chronic Periodontitis , Proteoglycans , Enzyme-Linked Immunosorbent Assay , Lymphocyte Function-Associated Antigen-1 , Gingival Crevicular Fluid , Intercellular Adhesion Molecule-1 , Neoplasm Proteins
5.
J. appl. oral sci ; 28: e20190558, 2020. graf
Article in English | LILACS, BBO | ID: biblio-1101249

ABSTRACT

Abstract Objective Ameloblastoma is a representative odontogenic tumor comprising several characteristic invasive forms, and its pathophysiology has not been sufficiently elucidated. A stable animal experimental model using immortalized cell lines is crucial to explain the factors causing differences among the subtypes of ameloblastoma, but this model has not yet been disclosed. In this study, a novel animal experimental model has been established, using immortalized human ameloblastoma-derived cell lines. Methodology Ameloblastoma cells suspended in Matrigel were subcutaneously transplanted into the heads of immunodeficient mice. Two immortalized human ameloblastoma cell lines were used: AM-1 cells derived from the plexiform type and AM-3 cells derived from the follicular type. The tissues were evaluated histologically 30, 60, and 90 days after transplantation. Results Tumor masses formed in all transplanted mice. In addition, the tumors formed in each group transplanted with different ameloblastoma cells were histologically distinct: the tumors in the group transplanted with AM-1 cells were similar to the plexiform type, and those in the group transplanted with AM-3-cells were similar to the follicular type. Conclusions A novel, stable animal experimental model of ameloblastoma was established using two cell lines derived from different subtypes of the tumor. This model can help clarify its pathophysiology and hasten the development of new ameloblastoma treatment strategies.


Subject(s)
Animals , Female , Mice , Ameloblastoma/pathology , Disease Models, Animal , Neoplasms, Experimental/pathology , Proteoglycans , Time Factors , Immunohistochemistry , Cells, Cultured , Reproducibility of Results , Collagen , Laminin , Cell Line, Tumor , Green Fluorescent Proteins/analysis , Drug Combinations
6.
Horiz. méd. (Impresa) ; 19(4): 84-92, Dic. 2019. tab, ilus
Article in Spanish | LIPECS, LILACS, LIPECS | ID: biblio-1048876

ABSTRACT

El glicocálix endotelial es una estructura sin forma definida que recubre la capa luminal del endotelio vascular y que está constituido, principalmente, por tres elementos: proteoglicanos, glucosaminoglicanos y glicoproteínas. Cumple distintas funciones, como regular la permeabilidad vascular a las moléculas y líquidos, la transducción de las fuerzas mecánicas de tensión y las cascadas de fibrinólisis y coagulación vascular; además, protege de la adhesión leucocitaria, plaquetaria y de patógenos. Los determinantes de lesión del glicocálix pueden ser de varios tipos, por ejemplo, incremento las fuerzas de tensión, especies reactivas de oxígeno (O2), aumento, a nivel plasmático, de sustancias como el sodio (hipernatremia), glucosa (hiperglicemia) y colesterol (hipercolesterolemia), y las moléculas proinflamatorias. Cualquiera de las noxas citadas, individualmente o combinadas, lesionan el glicocálix y la disfunción resultante se expresará clínicamente como disfunción endotelial, aumento de la permeabilidad vascular, paso de lipoproteínas al subendotelio, activación de la coagulación o aumento de la adhesión de plaquetas y leucocitos al endotelio.


Endothelial glycocalyx is an undefined structure covering the luminal layer of the vascular endothelium and consisting mainly of three elements: proteoglycans, glycosaminoglycans and glycoproteins. It has different functions, such as the regulation of vascular permeability to liquids and molecules; transduction of the mechanical forces of vascular tension; regulation of coagulation and fibrinolysis cascades; and protection of leukocyte, platelet and pathogen adhesion. The determinants of a glycocalyx lesion can be of several types­e.g., increased tensile forces; reactive oxygen (O2) species; increased plasma level of substances such as sodium (hypernatremia), glucose (hyperglycemia) and cholesterol (hypercholesterolemia); and pro-inflammatory molecules. Any of the above-mentioned noxas, alone or combined, injure the glycocalyx. Its dysfunction will be clinically expressed as endothelial dysfunction, increased vascular permeability, filtration of lipoproteins to the subendothelium, activation of coagulation, or increased adhesion of leukocytes and platelets to the endothelium.


Subject(s)
Humans , Glycocalyx , Proteoglycans , Endothelium
8.
Article in English | WPRIM | ID: wpr-763354

ABSTRACT

OBJECTIVE: This study was performed to explore the possibility that each oocyte and its surrounding cumulus cells might have different genetic expression patterns that could affect human reproduction. METHODS: Differential gene expression analysis was performed for 10 clusters of cumulus cells obtained from 10 cumulus-oocyte complexes from 10 patients. Same procedures related to oocyte maturation, microinjection, and microarray analyses were performed for each group of cumulus cells. Two differential gene expression analyses were performed: one for the outcome of clinical pregnancy and one for the outcome of live birth. RESULTS: Significant genes resulting from these analyses were selected and the top 20 affected pathways in each group were analyzed. Circadian entrainment is determined to be the most affected pathway for clinical pregnancy, and proteoglycans in cancer pathway is the most affected pathway for live birth. Circadian entrainment is also amongst the 12 pathways that are found to be in top 20 affected pathways for both outcomes, and has both lowest p-value and highest number of times found count. CONCLUSION: Although further confirmatory studies are necessary, findings of this study suggest that these pathways, especially circadian entrainment in cumulus cells, may be essential for embryo development and pregnancy.


Subject(s)
Circadian Clocks , Cumulus Cells , Embryonic Development , Female , Gene Expression , Granulosa Cells , Humans , Infertility , Live Birth , Microarray Analysis , Microinjections , Oocytes , Ovarian Follicle , Pregnancy , Proteoglycans , Reproduction , Reproductive Techniques, Assisted
9.
Article in English | WPRIM | ID: wpr-758954

ABSTRACT

The intra-articular use of hyaluronic acid (HA) for the treatment of synovitis and osteoarthritis is still controversial. As a consequence, corticosteroids remain the most frequently employed therapeutic agents, despite their potential systemic and local deleterious effects. This study examined the anti-inflammatory, antioxidant, and chondroprotective activities of low and high molecular weight hyaluronic acid (LMW-HA and HMW-HA) on lipopolysaccharide (LPS)-induced synovitis in horses compared to triamcinolone acetonide (TA). LPS was injected in the metacarpophalangeal joints, which were treated intra-articularly with either TA (as control) or LMW-HA or HMW-HA. Joint clinical evaluation and synovial fluid (SF) analysis were performed at 0, 8, 24, and 48 h. The white blood cell counts (WBC), prostaglandin E2 (PGE2), interleukin (IL)-1, IL-6, IL-10, tumor necrosis factor-α, chondroitin sulfate (CS) and HA concentrations, oxidative burst, and HA molecular weights were measured. TA reduced the lameness, swelling, and PGE2 release but increased the SF CS concentrations enormously at 24h and 48h, and decreased the SF HA modal molecular weight. These results indicate the breakdown of articular cartilage aggrecan and SF HA. In contrast, LMW-HA and HMW-HA were less effective in reducing the inflammation symptoms, but preserved the joints because only a modest increase in CS occurred at 24 h, decreasing at 48 h, and the SF HA was maintained. The HA-treatment also had anti-inflammatory actions, and LMW-HA was the most effective in reducing the release of cytokine. In summary, the HA treatment inhibited efficiently the digestion of cartilage proteoglycans and SF HA breakdown.


Subject(s)
Adrenal Cortex Hormones , Aggrecans , Cartilage , Cartilage, Articular , Chondroitin Sulfates , Digestion , Dinoprostone , Horses , Hyaluronic Acid , Inflammation , Interleukin-10 , Interleukin-6 , Interleukins , Joints , Leukocyte Count , Metacarpophalangeal Joint , Molecular Weight , Necrosis , Osteoarthritis , Proteoglycans , Respiratory Burst , Synovial Fluid , Synovitis , Triamcinolone , Triamcinolone Acetonide
10.
Article in English | WPRIM | ID: wpr-762680

ABSTRACT

PURPOSE: Transglutaminase type 2 (TG2) is an extracellular matrix crosslinking enzyme with a pivotal role in kidney fibrosis. We tested whether quantification of urinary TG2 may represent a noninvasive method to estimate the severity of kidney allograft fibrosis. METHODS: We prospectively collected urine specimens from 18 deceased donor kidney transplant recipients at 1-day, 7-day, 1-month, 3-month, and 6-month posttransplant. In addition, kidney allograft tissue specimens at 0-day and 6-month posttransplant were sampled to analyze the correlation of urinary TG2 and kidney allograft fibrosis. RESULTS: Thirteen recipients had increased interstitial fibrosis and tubular atrophy (IFTA) scores at the 6-month protocol biopsy (IFTA group). The mean level of urinary TG2 in the IFTA group was higher compared to that of 5 other recipients without IFTA (no IFTA group). Conversely, the mean level of urinary syndecan-4 in the IFTA group was lower than levels in patients without IFTA. In the IFTA group, double immunofluorescent staining revealed that TG2 intensity was significantly upregulated and colocalizations of TG2/heparin sulfate proteoglycan and nuclear syndecan-4 were prominent, usually around tubular structures. CONCLUSION: Urinary TG2 in early posttransplant periods is a potent biomarker for kidney allograft inflammation or fibrosis.


Subject(s)
Allografts , Atrophy , Biomarkers , Biopsy , Extracellular Matrix , Fibrosis , Humans , Inflammation , Kidney Transplantation , Kidney , Methods , Prospective Studies , Proteoglycans , Syndecan-4 , Tissue Donors , Transplant Recipients
11.
Article in English | WPRIM | ID: wpr-761902

ABSTRACT

BACKGROUND: This study aimed to observe the cartilaginous matrix production in SRY (sex determining region Y)-box 9 (SOX9)- and/or telomerase reverse transcriptase (TERT)-transfected chondrocytes from monolayer to three-dimensional (3D) culture.


Subject(s)
Alcian Blue , Cartilage , Chondrocytes , Clothing , Coloring Agents , Eosine Yellowish-(YS) , In Vitro Techniques , Proteoglycans , Real-Time Polymerase Chain Reaction , Regeneration , Telomerase , Tissue Engineering , Transfection
12.
Int. j. morphol ; 37(1): 54-58, 2019. graf
Article in English | LILACS | ID: biblio-990004

ABSTRACT

SUMMARY: Matrigel is a basement membrane matrix extracted from the EHS mouse tumor containing extracellular matrix protein, its main components are laminin, type IV collagen, nestin, heparin sulfate, growth factor and matrix metalloproteinase.At room temperature, Matrigel polymerized to form a three dimensional matrix with biological activity. It can simulate the structure, composition, physical properties and functions of the cell basement membrane in vivo, which is beneficial to the culture and differentiation of the cells in vitro, and can be used for the study of cell morphology, biochemical function, migration, infection and gene expression. In this study, Matrigel three-dimensional culture model of bone marrow mesenchymal stem cells(BMSCs) was established, and its morphology, proliferation and survival were observed. BMSCs were isolated and cultured with whole bone marrow adherence method. The Second generation BMSCs with good growth condition were selected and mixed with Matrigel to form cell gel complexes. The morphology and proliferation of mesenchymal stem cells were observed by phase contrast microscope and HE staining,Live/Dead staining was used to evaluate the cell activity.Phase contrast microscopy showed that BMSCs were reticulated in Matrigel and proliferated well, After 7 days, the matrix gel gradually became soft and collapsed, a few cell reticular crosslinking growth was seen at 14 days; HE staining showed that the cytoplasm of the cells was larger on the fourth day and the cells were elongated and cross-linked on the seventh day; Live/dead staining showed that most cells showed green fluorescence with the prolongation of culture time, on the first, 4 and 7 days, the activity of bone marrow mesenchymal stem cells in Matrigel gradually increased, and the percentages were 92.57 %, 95.54 % and 97.37 %, respectively. Matrigel three-dimensional culture system can maintain the morphology, function and proliferation ability of bone marrow mesenchymal stem cells.


RESUMEN: Matrigel es una matriz de membrana basal extraída del tumor de ratón EHS que contiene proteína de matriz extracelular. Los componentes principales son laminina, el colágeno tipo IV, nestina, sulfato de heparina, factor de crecimiento y metaloproteinasa de matriz. A temperatura ambiente, Matrigel se polimerizó para formar una matriz tridimensional. Es posible simular la estructura, la composición, las propiedades físicas y las funciones de la membrana basal celular in vivo, lo que es beneficioso para el cultivo y la diferenciación de las células in vitro, y se puede utilizar para el estudio de la morfología celular, la función bioquímica, la migración, infección y expresión génica. En este estudio, se estableció el modelo de cultivo tridimensional Matrigel de células madre mesenquimales de médula ósea (BMSC), y se observó su morfología, proliferación y supervivencia. Las BMSC fueron aisladas y cultivadas con el método de adherencia de la médula ósea completa. Se seleccionaron las BMSC de segunda generación con buenas condiciones de crecimiento y se mezclaron con Matrigel para formar complejos de gel de células. La morfología y la proliferación de las células madre mesenquimales se observaron con microscopio de contraste de fase y se tiñó con Hematoxilina-Eosina (HE); para evaluar la actividad celular se usó la tinción Live/Dead. La microscopía de contraste mostró que las BMSC se reticularon en Matrigel y proliferaron bien. Después de 7 días, se observó que el gel de matriz gradualmente se volvió blando y colapsó, y se visualizó un cruce transversal de algunas células reticulares a los 14 días. La tinción mostró que la mayoría de las células mostraron una fluorescencia verde con la prolongación del tiempo de cultivo; en los primeros 4 y 7 días, la actividad de las células madre mesenquimales de la médula ósea en Matrigel aumentó gradualmente y los porcentajes fueron de 92,57 %, 95,54 % y 97,37 %, respectivamente. El sistema de cultivo tridimensional de Matrigel puede mantener la morfología, la función y la capacidad de proliferación de las células madre mesenquimales de la médula ósea.


Subject(s)
Animals , Dogs , Proteoglycans/chemistry , Collagen/chemistry , Laminin/chemistry , Cell Culture Techniques/methods , Mesenchymal Stem Cells/cytology , Tissue Engineering , Drug Combinations
13.
Braz. oral res. (Online) ; 33: e059, 2019. graf
Article in English | LILACS | ID: biblio-1039303

ABSTRACT

Abstract We recently demonstrated that a co-culture system of human umbilical vein endothelial cells (HUVECs) and human dental pulp stem cells (hDPSCs) could enhance angiogenesis ability in vitro. However, whether tumor necrosis factor α (TNF-α) could promote blood vessel formation during pulp regeneration remained unknown. The aim of this study was to investigate the effects of TNF-α on the formation of endothelial tubules and vascular networks in a co-culture system of hDPSCs and HUVECs. hDPSCs were co-cultured with HUVECs at a ratio of 1:5. The Matrigel assay was performed to detect the total tubule branching lengths and numbers of branches, and the Cell-Counting Kit 8 assay was performed to examine the effect of TNF-α on cell proliferation. Real-time polymerase chain reactions and western blot were used to detect vascular endothelial growth factor (VEGF) mRNA and protein expression. The Matrigel assay showed significantly greater total branching lengths and numbers of branches formed in the experimental groups treated with different concentrations of TNF-α compared with the control group. The decomposition times of the tubule structures were also significantly prolonged (P < 0.05). Treatment with 50 ng/ml TNF-α did not significantly change the proliferation of co-cultured cells, but it significantly increased the VEGF mRNA and protein expression levels (p < 0.05). In addition, the migration abilities of HUVECs and hDPSCs increased after co-culture with TNF-α (p < 0.05). TNF-α enhanced angiogenic ability in vitro in the co-culture system of hDPSCs and HUVECs.


Subject(s)
Humans , Adolescent , Adult , Young Adult , Tumor Necrosis Factor-alpha/pharmacology , Neovascularization, Physiologic/drug effects , Dental Pulp/cytology , Dental Pulp/drug effects , Angiogenesis Inducing Agents/pharmacology , Human Umbilical Vein Endothelial Cells/drug effects , Proteoglycans , Reference Values , Time Factors , Cell Count , Cells, Cultured , Blotting, Western , Reproducibility of Results , Collagen , Laminin , Neovascularization, Physiologic/physiology , Dental Pulp/physiology , Vascular Endothelial Growth Factor A/analysis , Vascular Endothelial Growth Factor A/drug effects , Cell Proliferation/drug effects , Cell Proliferation/physiology , Drug Combinations , Cell Migration Assays , Human Umbilical Vein Endothelial Cells/physiology , Real-Time Polymerase Chain Reaction
14.
Rev. bras. ginecol. obstet ; 40(10): 593-598, Oct. 2018. tab, graf
Article in English | LILACS | ID: biblio-977779

ABSTRACT

Abstract Objective To analyze endocan-1, a biomarker of vascular endothelial related pathologies, and the placental growth factor (PlGF), an angiogenic factor and a placental dysfunction marker in patients with preeclampsia (PE). Methods Case-control study conducted at Hospital São Lucas, in the city of Porto Alegre, Brazil. Endocan-1 and PlGF levels were quantified in the maternal plasma using the MagPlexTH-C microsphere system (MAGPIX System, Luminex, Austin, Texas, US) and evaluated through analysis of covariance (ANCOVA) and adjusted by body mass index (BMI), gestational age and maternal age. To estimate the difference between the groups, the mean ratio (MR) and the 95% confidence interval (95%CI) were calculated. The Pearson correlation test was used to establish any association between endocan-1 and PlGF levels. The null hypothesis was rejected when p < 0.05. Results The group of patients was composed by normotensive (n = 67) patients and patients with PE (n = 50). A negative correlation between endocan-1 and the PlGF was noted in the entire normotensive group (linear correlation coefficient [r] = -0.605; p < 0.001), as well as in the PE group (r = -0.545; p < 0.001). Conclusion Endocan-1 levels are increased in patients with PE, and are inversely correlated with PlGF levels. We suggest that it is important to analyze angiogenic and proinflammatory molecules concomitantly in women with PE to better understand the pathophysiology of the disease. Both molecules are strong candidates for PE biomarkers, and future studies will examine any mechanisms connecting these factors in PE.


Resumo Objetivo Analisar o endocan-1, umbiomarcador de patologias vasculares endoteliais, e o fator de crescimento placentário (FCPl), um fator angiogênico, marcador de disfunção placentária em pacientes com pré-eclâmpsia (PE). Métodos Estudo de caso-controle realizado no Hospital São Lucas, em Porto Alegre. Os níveis de endocan-1 e FCPl foram quantificados no plasma materno usando o sistema de microesferas MagPlexTH-C (MAGPIX System, Luminex, Austin, Texas, US) e analisados por análise de covariância (ANCOVA) e ajustados por índice de massa corporal (IMC), idade gestacional e idade materna. Para calcular a diferença entre os grupos, utilizou-se a razão dasmédias (RM) e o intervalo de confiança de 95% (IC95%). O teste de correlação de Pearson foi utilizado para estabelecer a associação entre os níveis de endocan-1 e FCPl. A hipótese nula foi rejeitada quando p < 0,05. Resultados O grupo de pacientes foi composto por pacientes normotensas (n = 67) e pacientes com PE (n = 50). Uma correlação negativa entre o endocan-1 e o FCPl foi observada emtodo o grupo de pacientes normotensas (coeficiente de correlação linear [r] = -0,605; p < 0,001), bem como no grupo com PE (r = -0,545; p < 0,001). Conclusão Os níveis de endocan-1 estão aumentados em pacientes com PE e inversamente correlacionados com os níveis de FCPl. Sugerimos a importância de analisar moléculas angiogênicas e pró-inflamatórias concomitantemente em mulheres com PE para compreender melhor a fisiopatologia da doença. Ambas as moléculas são fortes candidatos a serem considerados biomarcadores de PE, e trabalhos futuros poderão avaliar quaisquer mecanismos que liguem esses fatores na PE.


Subject(s)
Humans , Female , Adult , Pre-Eclampsia/blood , Proteoglycans/blood , Placenta Growth Factor/blood , Neoplasm Proteins/blood , Biomarkers/blood , Case-Control Studies , Correlation of Data
15.
Article in English | WPRIM | ID: wpr-776071

ABSTRACT

OBJECTIVE@#Newly identified human rhinovirus C (HRV-C) and human bocavirus (HBoV) cannot propagate in vitro in traditional cell culture models; thus obtaining knowledge about these viruses and developing related vaccines are difficult. Therefore, it is necessary to develop a novel platform for the propagation of these types of viruses.@*METHODS@#A platform for culturing human airway epithelia in a three-dimensional (3D) pattern using Matrigel as scaffold was developed. The features of 3D culture were identified by immunochemical staining and transmission electron microscopy. Nucleic acid levels of HRV-C and HBoV in 3D cells at designated time points were quantitated by real-time polymerase chain reaction (PCR). Levels of cytokines, whose secretion was induced by the viruses, were measured by ELISA.@*RESULTS@#Properties of bronchial-like tissues, such as the expression of biomarkers CK5, ZO-1, and PCK, and the development of cilium-like protuberances indicative of the human respiration tract, were observed in 3D-cultured human airway epithelial (HAE) cultures, but not in monolayer-cultured cells. Nucleic acid levels of HRV-C and HBoV and levels of virus-induced cytokines were also measured using the 3D culture system.@*CONCLUSION@#Our data provide a preliminary indication that the 3D culture model of primary epithelia using a Matrigel scaffold in vitro can be used to propagate HRV-C and HBoV.


Subject(s)
Collagen , Drug Combinations , Enterovirus , Enterovirus Infections , Virology , Enzyme-Linked Immunosorbent Assay , Epithelial Cells , Virology , Human bocavirus , Humans , Laminin , Parvoviridae Infections , Virology , Primary Cell Culture , Methods , Proteoglycans , Real-Time Polymerase Chain Reaction , Respiratory Mucosa , Virology , Virus Cultivation
16.
Article in Chinese | WPRIM | ID: wpr-813189

ABSTRACT

To determine the effect of andrographolide (Andro) on angiogenesis of human umbilical vein endothelial cells (HUVECs).
 Methods: HUVECs were treated with different concentrations of Andro and the cell viability was detected with Cell Counting Kit-8 (CCK-8). HUVECs were treated with half lethal dose (IC50) of Andro. Matrigel was used to make capillary formation of HUVECs and the effect of Andro on capillary formation was evaluated by calculating the percentage of capillary formation. Moreover, the effects of Andro and the supernatant from cultured A549 tumor cells on capillary formation were evaluated by calculating the percentage of capillary formation. The effect of Andro on the expression of matrix metalloproteinase-9 (MMP-9) was determined with Western blot.
 Results: The cell viability of HUVECs decreased with the increase of Andro concentrations. IC50 was 20 μmol/L. The capillary formation of HUVECs was inhibited when treated with 20 μmol/L Andro for 24 hours. Moreover, Andro was able to antagonize the promotion of the capillary formation induced by the supernatant from cultured tumor cells. Andro could suppress the expression of MMP-9 and antagonize the capillary formation.
 Conclusion: Andro inhibits the capillary formation of HUVECs and can antagonize the promotion of angiogenesis induced by the supernatant from cultured tumor cells.


Subject(s)
Capillaries , Cell Survival , Collagen , Culture Media , Diterpenes , Pharmacology , Drug Combinations , Human Umbilical Vein Endothelial Cells , Humans , Laminin , Matrix Metalloproteinase 9 , Metabolism , Neovascularization, Pathologic , Proteoglycans , Tumor Cells, Cultured
17.
Article in English | WPRIM | ID: wpr-719105

ABSTRACT

Aggrecan is a proteoglycan in the extracellular matrix of growth plate and cartilaginous tissues. Aggrecanopathy has been reported as a genetic cause not only for severe skeletal dysplasia but also for autosomal dominant short stature with normal to advanced bone age. We report a novel heterozygous mutation of ACAN in a Korean family with proportionate short stature identified through targeted exome sequencing. We present a girl of 4 years and 9 months with a family history of short stature over three generations. The paternal grandmother is 143 cm tall (−3.8 as a Korean standard deviation score [SDS]), the father 155 cm (−3.4 SDS), and the index case 96.2 cm (−2.9 SDS). Evaluation for short stature showed normal growth hormone (GH) peaks in the GH provocation test and a mild delayed bone age for chronological age. This subject had clinical characteristics including a triangular face, flat nasal bridge, prognathia, blue sclerae, and brittle teeth. The targeted exome sequencing was applied to detect autosomal dominant growth palate disorder. The novel variant c.910G>A (p.Asp304Asn) in ACAN was identified and this variant was found in the subject's father using Sanger sequencing. This is the first case of Korean familial short stature due to ACAN mutation. ACAN should be considered for proportionate idiopathic short stature, especially in cases of familial short stature.


Subject(s)
Aggrecans , Exome , Extracellular Matrix , Family Characteristics , Fathers , Female , Grandparents , Growth Hormone , Growth Plate , Humans , Palate , Proteoglycans , Sclera , Tooth
18.
Article in English | WPRIM | ID: wpr-718787

ABSTRACT

BACKGROUND: Glucosamine hydrochloride (GlcN·HCl) has been shown to inhibit cell growth and matrix synthesis, but not with N-acetyl-glucosamine (GlcNAc) supplementation. This effect might be related to an inhibition of critical growth factors (GF), or to a different metabolization of the two glucosamine derivatives. The aim of the present study was to evaluate the synergy between GlcN·HCl, GlcNAc, and GF on proliferation and cartilage matrix synthesis. METHOD: Bovine chondrocytes were cultivated in monolayers for 48 h and in three-dimensional (3D) chitosan scaffolds for 30 days in perfusion bioreactors. Serum-free (SF) medium was supplemented with either growth factors (GF) TGF-β (5 ng mL₋₁) and IGF-I (10 ng mL₋₁), GlcN·HCl or GlcNAc at 1mM each or both. Six groups were compared according to medium supplementation: (a) SF control; (b) SF + GlcN·HCl; (c) SF + GlcNAc; (d) SF + GF; (e) SF + GF + GlcN·HCl; and (f) SF + GF + GlcNAc. Cell proliferation, proteoglycan, collagen I (COL1), and collagen II (COL2) synthesis were evaluated. RESULTS: The two glucosamines showed opposite effects in monolayer culture: GlcN·HCl significantly reduced proliferation and GlcNAc significantly augmented cellular metabolism. In the 30 days 3D culture, the GlcN·HCl added to GF stimulated cell proliferation more than when compared to GF only, but the proteoglycan synthesis was smaller than GF. However, GlcNAc added to GF improved the cell proliferation and proteoglycan synthesis more than when compared to GF and GF/GlcN·HCl. The synthesis of COL1 and COL2 was observed in all groups containing GF. CONCLUSION: GlcN·HCl and GlcNAc increased cell growth and stimulated COL2 synthesis in long-time 3D culture. However, only GlcNAc added to GF improved proteoglycan synthesis.


Subject(s)
Bioreactors , Cartilage , Cell Proliferation , Chitosan , Chondrocytes , Collagen , Glucosamine , Insulin-Like Growth Factor I , Intercellular Signaling Peptides and Proteins , Metabolism , Methods , Perfusion , Proteoglycans
19.
Protein & Cell ; (12): 298-309, 2018.
Article in English | WPRIM | ID: wpr-756940

ABSTRACT

The blood-brain barrier (BBB) is a tight boundary formed between endothelial cells and astrocytes, which separates and protects brain from most pathogens as well as neural toxins in circulation. However, detailed molecular players involved in formation of BBB are not completely known. Dentin matrix protein 1 (DMP1)-proteoglycan (PG), which is known to be involved in mineralization of bones and dentin, is also expressed in soft tissues including brain with unknown functions. In the present study, we reported that DMP1-PG was expressed in brain astrocytes and enriched in BBB units. The only glycosylation site of DMP1 is serine89 (S89) in the N-terminal domain of the protein in mouse. Mutant mice with DMP1 point mutations changing S89 to glycine (S89G), which completely eradicated glycosylation of the protein, demonstrated severe BBB disruption. Another breed of DMP1 mutant mice, which lacked the C-terminal domain of DMP1, manifested normal BBB function. The polarity of S89G-DMP1 astrocytes was disrupted and cell-cell adhesion was loosened. Through a battery of analyses, we found that DMP1 glycosylation was critically required for astrocyte maturation both in vitro and in vivo. S89G-DMP1 mutant astrocytes failed to express aquaporin 4 and had reduced laminin and ZO1 expression, which resulted in disruption of BBB. Interestingly, overexpression of wild-type DMP1-PG in mouse brain driven by the nestin promoter elevated laminin and ZO1 expression beyond wild type levels and could effectively resisted intravenous mannitol-induced BBB reversible opening. Taken together, our study not only revealed a novel element, i.e., DMP1-PG, that regulated BBB formation, but also assigned a new function to DMP1-PG.


Subject(s)
Animals , Astrocytes , Cell Biology , Metabolism , Blood-Brain Barrier , Cell Biology , Metabolism , Cells, Cultured , Extracellular Matrix Proteins , Genetics , Metabolism , Female , Glycosylation , Male , Mice , Proteoglycans , Metabolism , Reverse Transcriptase Polymerase Chain Reaction
20.
Article in Chinese | WPRIM | ID: wpr-690811

ABSTRACT

<p><b>OBJECTIVE</b>To observe the impacts of thermosensitive moxibustion (TSM) on the expressions of nitric oxide (NO), typeⅠdisintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), typeⅡcollagen and proteoglycan (PG) in the rabbit models of knee osteoarthritis (KOA) and explore the mechanism of TSM on KOA.</p><p><b>METHODS</b>A total of 42 Japanese long-eared male rabbits were divided into a blank group (6 rabbits), a model group (6 rabbits), a moxibustion group (24 rabbits) and a sham-operation group (6 rabbits) according to the random number table. In the blank group, the rabbits were fed normally. In the model and moxibustion groups, the papain injection was given to establish KOA models. The rabbits in the sham-operation group were treated with the intracavity injection of 0.9% NaCl solution. The rabbits were forced to move for 30 min every day, continuously for 15 days during modeling. At the end of modeling, in the moxibustion group, moxibusiton was applied at "Dubi" (ST 35), once a day, 40 min each time, for 14 days totally. According to the temperature changes during moxibustion, the rabbits were divided into a TSM group and a non-TSM group. 6 rabbits were collected randomly from the two groups. The usual feeding was given in the blank group, the model group and the sham-operation group, without any intervention. The body mass and behavioristics changes were observed in each group. At the end of treatment, the nitrate reduction method was adopted to determine NO expression in the serum. The real-time PCR was adopted to determine the expressions of ADAMTS-4, typeⅡcollagen and PG in the cartilage.</p><p><b>RESULTS</b>① After modeling, compared with the blank group, the body mass was all reduced and the Lequesne MG score was increased in the model group, TSM group, non-TSM group and sham-operation group (<0.05, <0.01). After intervention, compared with the blank group, the body mass was decreased and the Lequesne MG score was increased in the model and sham-operation groups (<0.05, <0.01). Compared with the model group, the body mass was increased and the lequesne MG score was decreased in the TSM, non-TSM, and sham-operation groups (<0.05, <0.01). Compared with the non-TSM group, the body mass in the TSM group was increased remarkably (<0.05), but the difference in Lequesne MG score was not statistically significant (>0.05). ② After intervention, compared with the blank group, the expressions of NO and ADAMTS-4 were all increased and the expressions of typeⅡcollagen and PG were decreased in the model group, TSM group, non-TSM group and sham-operation group (<0.05, <0.01). Compared with the model group, the expressions of NO and ADAMTS-4 were all remarkably lower and the expressions of typeⅡcollagen and PG were increased in the TSM group, non-TSM group and sham-operation group (<0.05, <0.01). Compared with the non-TSM group, the expressions of NO and ADAMTS-4 were all remarkably lower and the expressions of typeⅡcollagen and PG were increased in the TSM group after intervention (all <0.05).</p><p><b>CONCLUSION</b>The thermosensitive moxibustion alleviates the inflammatory reactions and protects the joint cartilage through inhibiting the expressions of NO and ADAMTS-4 to achieve the effects in the treatment of KOA.</p>


Subject(s)
ADAMTS4 Protein , Metabolism , Animals , Cartilage , Metabolism , Collagen Type III , Metabolism , Male , Moxibustion , Nitric Oxide , Blood , Osteoarthritis, Knee , Therapeutics , Proteoglycans , Metabolism , Rabbits , Random Allocation
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