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ABSTRACT This study aimed to analyze the anterior lens capsule specimens from both eyes of a patient with systemic sclerosis and compare them to the eyes of a control patient. No significant differences between systemic sclerosis and control eyes were observed in the results from the hematoxylin-eosin and picrosirius staining. In the samples obtained from both systemic sclerosis and control eyes, there were expressions of caspase, a molecule expressed in cell death by apoptosis. Heparanase was overexpressed in the systemic sclerosis sample compared to the control sample. Therefore, the anterior lens capsule of the patient with systemic sclerosis is probably affected by the disease since it showed marked expression of heparanase 1.(AU)
RESUMO Analisamos as amostras das cápsulas anteriores do cristalino de uma paciente com esclerose sistêmica e comparamos com as de um paciente controle. Não foram observadas diferenças significativas entre esclerose sistêmica e controle nos resultados da coloração com hematoxilina-eosina e picrosirius. Nas amostras obtidas da esclerose sistêmica e do controle, obtivemos expressão de caspase, uma molécula expressa na morte celular por apoptose. A heparinase foi expressa de forma mais marcante na amostra de esclerose sistêmica quando comparada ao controle. Portanto, a cápsula anterior do cristalino da paciente com esclerose sistêmica provavelmente foi afetada pela doença, uma vez que mostrou expressão aumentada de heparinase 1.(AU)
Subject(s)
Humans , Scleroderma, Systemic/physiopathology , Heparin Lyase/administration & dosage , Hematoxylin , Lens Capsule, Crystalline/anatomy & histologyABSTRACT
Objective To evaluate the role of heparanase in endotoxemia-induced destruction of vascular endothelial glycocalyx in rats.Methods Forty-eight pathogen-free male Sprague-Dawley rats, aged 8-10 weeks, weighing 200-250 g, were randomly divided into 3 groups (n=16 each) using a random number table: control group (group C), lipopolysaccharide (LPS) group (group L) and LPS+ heparin group (group LH).Endotoxemia was induced by intravenous LPS 15 mg/kg in L and LH groups, and heparin sodium 100 U · kg-1 · h-1 was infused simultaneously in group LH.At 3 and 6 h after LPS injection, blood samples were collected from the femoral vein for determination of the serum concentrations of serum heparin sulfate (HS), syndecan-1, E-selectin and intercellular adhesion molecule-1 (ICAM-1) concentrations.The rats were then sacrificed, and lungs were removed for microscopic examination and for determination of wet/dry lung weight ratio (W/D ratio).Results Compared with group C, the serum HS, syndecan-1, E-selectin and ICAM-1 concentrations and W/D ratio were significantly increased in group L, and the serum HS and ICAM-1 concentrations were increased in group LH (P＜0.05).Compared with group L, the serum HS, syndecan-1, E-selectin and ICAM-1 concentrations and W/D ratio were significantly decreased in group LH (P＜0.05).The pathological changes of lungs were obvious in group L, and were significantly mitigated in group LH.Conclusion Heparanase is involved in endotoxemiainduced destruction of vascular endothelial glycocalyx in rats.
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Objective To evaluate the effects of sevoflurane on the expression of heparanase ( HPA) and fascin in lung carcinoma cells of mice. Methods Mouse LLC cells were inoculated in the culture plate. After being cultured for 24 h, the cells were equally and randomly divided into 4 groups using a random number table: control group ( group CC) , 1% sevoflurane group ( group Sev1 ) , 2% sevoflurane group ( group Sev2 ) , and 3% sevoflurane group ( group Sev3 ) . Cells in Sev1-3 groups were exposed to 1%, 2% and 3% sevoflurane, respectively, for 4 h, while cells in group CC were not exposed to sevoflurane, and all the cells were then cultured for another 24 h in an incubator. The invasion of cells was determined by Transwell invasion assay, and the invaded cells were counted. The migration of cells was determined by wound healing assay, and cell migration rates were calculated. The expression of HPA and fascin in cells was detected by Western blot. Results Compared with group CC, the number of invaded cells and cell migration rates were gradually decreased, and the expression of HPA and fascin was gradually down?regulated with increasing concentrations of sevoflurane in Sev1-3 groups. Conclusion The mechanism through which sevoflurane inhibits the metastasis of mouse lung carcinoma cells is associated with down?regulated expression of HPA and fascin.
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BACKGROUND:Accumulating evidence shows that, low molecular heparin can prevent and treat early osteonecrosis of femoral head, but the mechanism remains unclear. The heparin exhibits a dynamic balance with heparanase in vivo, and inhibits heparanase expression. OBJECTIVE:To observe the changes of heparanase and tissue factor pathway inhibitor expression level in plasma and femoral head of patients with idiopathic osteonecrosis of femoral head, and investigate the pathophysiological role of heparanase in the progression of idiopathic osteonecrosis of femoral head. METHODS:A total of 36 patients with idiopathic osteonecrosis of femoral head were randomly divided into four groups according to ARCO I-IV period, with nine patients in each group. Patients with fresh femoral neck fracture for hip arthroplasty were included in the control group. Heparanase and tissue factor pathway inhibitor levels in preoperative venous plasma were measured by ELISA, and in subchondral necrosis bone of ARCO II-IV groups were observed for western blot analysis and hematoxylin-eosin staining. RESULTS AND CONCLUSION:In plasma, heparanase and tissue factor pathway inhibitor levels were significantly increased along with the development of idiopathic osteonecrosis of femoral head compared with control group, the highest levels were found in ARCO III period, and then slightly decreased in ARCO IV period, showing significant differences. The protein levels of heparanase and tissue factor pathway inhibitor in necrosis bone were increased significantly compared with control group, the highest levels occurred in ARCO III period, no obvious difference was found between ARCO II and IV periods. Heparanase and tissue factor pathway inhibitor levels are gradual y increased along with the development of the necrosis, and heparanase plays an important role in idiopathic osteonecrosis of femoral head.
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Objective To investigate the CD44v6 and heparanase in non-small cell lung cancer (NSCLC ) and adjacent normal lung tissues ,and to study the correlation of these two molecular in non-small cell lung cancer invasion and metastasis .Methods The expressing of CD44v6 and heparanase in 117 patients with non-small cell lung cancer tissues and 62 adjacent normal lung tissue was detected by immunohistochemical staining .Results The expression of CD44v6 in NSCLC was 54 .7% (64/117) ,which was sig-nificantly higher than in adjacent tissues of 4 .8% (3/62)(P<0 .05);the expression of heparanase in NSCLC was 86 .3% (101/117) ,which was significantly higher than the normal adjacent tissues 17 .7% (11/62)(P<0 .01);CD44v6 expression in NSCLC ex-pression was positive correlated with heparanase .Conclusion CD44v6 and heparanase are associated with the invasion and metasta-sis of NSCLC ,combined detection of the expression of the two molecular maybe has a certain reference value in determine the prog-nosis of NSCLC .
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Objective To observe the impact of heparanase on glomerular endothelium glycocalyx during sepsis and to investigate the prevention of glycocalyx injury.Methods C57/BL6 mice were injected with lipopolysaccharide (LPS) or tumor necrosis factor-α(TNF-o) and sacrificed one hour later.Glomerular endothelium glycocalyx traced with lanthanum was observed by transmission electron microscope(TEM).Western blotting was used to observe heparanse protein expression of renal cortex tissue.Human renal glomerular endothelial cells (HRGECs) were stimulated with TNF-α and active heparanase protien expression was detected by Western blotting.Mice were administrated with heparin sodium or heparinase Ⅲ and renal endothelium glycocalyx was observed by TEM.Urine during twenty-four hours was collected to measure urinary albumin and creatinine.The ratio of albumin to creatinine was calculated and compared among groups.Results The glomerular endothelium glycocalyx of LPS group and TNF-α group was degradated and the one of podocyte was integrated.Renal cortex tissue heparanase protein expression was significantly increased since one hour after LPS injection (P ＜ 0.01).The protein expression of activited heparanase of HRGECs which were stimulated with TNF-α was increased (P ＜ 0.05).Administration of heparin sodium which could inhibit the activity of heparanase could prevent the glycocalyx form degradation.The ratio of urine albumin to creatinine of heparin sodium group was decreased compared with LPS group (P ＜ 0.05) and the ratio of heparinase Ⅲ group was higher than control group(P ＜ 0.01) as a result of degradation of glomerular endothelium glycocalyx.Conclusions During the early stage of sepsis,TNF-α can induce glomerular endothelium heparanase to increase and active,and consequently the glycocalyx is degradated which leads to albuminuria.Inhibition of heparanase can protect glomerular endothelium glycocalyx and prevent albuminuria.
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Objective To investigate the relationship between the expression of DEK and Hpa infection in uterine cervical carcinogenisis.Methods Formalin-fixed,paraffin-embedded samples including 45 cases of squamous cell carcinomas,25 cases of cervical intraepithelial neoplasia CINⅢ,and 20 cases of CINI-Ⅱ,10 cases of normal cervical tissues were studied using polymer detection system for immunohistochemical technique to examine the expression of DEK and Hpa.Results The positive ratios of DEK in cancer tissues,normal cervical intraepithelial lesions,normal cervical tissues positive rates were 88.9％,24.4％,10.0％,respectively.The positive ratio of expressions of DEK in cervical carcinomas was significantly higher than CIN and normal cervical tissues (x2 =38.054,P ＜0.01 ;x2 =5.774,P ＜0.05).The positive ratios of expressions of Hpa in normal cervical tissues,CIN and cervical cancers were 75.6％,24.4％ and 10.0％,respectively.The positive ratio of expressions of Hpa in cervical carcinomas was significantly higher than CIN and normal cervical tissues ( x2 =31.69,P ＜ 0.01 ; x2 =12.94,P ＜ 0.01 ).The expression of DEK and Hpa in cervical lesions was highly related in the cervical cancer tissues( r =0.617,P =0.001 ).Conclusions DEK and Hpa possibly played an important role in the development of cervical cancer.Cervical intraepithelial lesions and cervical carcinoma might be associated with changes in the expression of DEK and Hpa,and interact together to influence the development of CIN and cervical cancer.
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AIM: To determine whether the eukaryotic initiation factor-4E (eIF-4E) inhibition facilitates the degradation of heparanase mRNA and alters heparanase protein expression in human colon adenocarcinoma cell line, LS-174T. METHODS: A 20-mer antisense s-oligodeoxynucleotide (asODN) targeted against the translation start site of eIF-4E mRNA were introduced into LS-174T cells by lipid-mediated DNA-transfection. eIF-4E protein and mRNA levels were detected by Western blot and RT-PCR, respectively. The mRNA levels of heparanase were determined by Northern blot. The alterations of heparanase expression were confirmed by Western blot analysis. RESULTS: The 20-mer asODN against eIF-4E specifically and significantly inhibited eIF-4E protein expression. Following eIF-4E inhibition, a significant reduction of heparanase mRNA was observed on Northern blot, and at the same time, heparanase protein expression significantly decreased as well. CONCLUSIONS: The results indicate that the inhibition of eIF-4E strongly reduces the stability of heparanase mRNA in colon adenocarcinoma cell line, LS-174T and resultes in an apparent reduction in the expression of heparanase protein. [