Objective To study the change in serum levels of soluble CD14, tumor necrosis factor-α, E-selectin, interleukin-10 and mean arterial pressure, as well as their relationship to infection during the pathophysiologic process in endotoxemia of rabbits. Methods Sixteen rabbits were randomly divided into two groups: group A, as a control group; group B, endotoxemia group. The model of rabbit with endotoxemia were used. Endotoxin at a dose of 1.5 mg/(kg·h) or 3 mg/(kg·h) was continuously infused through external jugular vein within 2 hours, 1 hour respectively. The change of levels of serum soluble CD14, tumor necrosis factor-α, interleukin-10 and E-selectin were observed at 0 (time before infusion of endotoxin), 30, 60, 120, 180, 240, 300 minutes, while mean arterial pressure was measured by polygraphy system. Results In the group B,there was an increase of content of soluble CD14,tumor necrosis factor-α,interleukin-10 and E-selectin following 30, 120 minutes respectively,and mean arterial pressure was lower than that of group A at same time points. Conclusion The results suggest that soluble CD14,tumor necrosis factor-α,interleukin-10 and E-selectin may play an important role during the change of infection and that these changes may be closely related with severe infection.
To evaluate the effect of intercellular adhesiveness molecule-1 (ICAM-1), E-selectin on hepatic microcirculation in acute cholangitis. The Changes of hepatic tissue, content of blood flow and Evan′s blue (EB) in hepatic tissue in acute cholangitis were determinated. Results: The number of PMN in hepatic tissue and sinusin increased, degenaration and necrosis of the hepatic cells and hepatic sinusoidal endothelial cells and content of blood flow in liver were reduced, and content of EB in hepatic tissue increased remarkbly in the rats with acute cholangitis. Pretreatment of anti ICAM-1 and E-selectin mAb reduced the damage of hepatic microcirculation. Conclusion: ICAM-1 and E-selectin may play an important role in damage to hepatic microcirculation in acute cholangitis.
Objective To study the relationship between smoking as well as smoke cessation and lung inflammation of chronic obstructive pulmonary disease( COPD) , and its possible mechanism. Methods Thirty-two Wistar rats were randomly divided into a control group, a low-dose smoking group, a high-dose smoking group, and a smoke cessation group. The expressions of NF-κB and E-selectin in rats lung were detected by immunohistochemistry and hybridization in situ. Results The mRNA and protein expressions of NF-κB in bronchial endothelial cells of the high-dose smoking group( 0. 43 ±0. 01, 0. 41 ±0. 04) , the lowdose smoking group( 0. 41 ±0. 01, 0. 40 ±0. 01) , and the smoke cessation group( 0. 39 ±0. 01, 0. 37 ±0. 03) , were significantly higher than the control group( 0. 29 ±0. 06, 0. 28 ±0. 06) ( all P lt; 0. 05) , with the high-dose smoking group and the smoke cessation group significantly higher than the low-dose smoking group ( both P lt;0. 05) . The mRNA and protein expressions of E-selectin had the similar pattern with NF-κB. The expressions of E-selectin mRNA and protein were positively correlated with the expression of NF-κB mRNA and protein respectively ( r = 0. 80, r = 0. 89, P lt; 0. 01) . Conclusions Smoking can result in high expressions of NF-κB and E-selectin in a dose dependent manner. Smoke cessation can relieve airway inflammation and is an effective measure for preventing COPD.
Abstract: Objective To study the expression of E-selectin on vascular endothelial cells of nude mice liver induced by esophageal carcinoma cells, in order to find out the function of E-selectin in the metastasis of esophageal carcinoma into the liver. Methods Twelve Balb/c nude mice aged from 6 to 8 weeks with their weight ranged between 20 and 25 grams were selected in our research. The mice were equally distributed into the experimental group and the control group(n=6). EC9706 cell solution (5×10.6/0.02 ml) were injected beneath the splenic capsule of the mice in the experimental group. One hour later, spleen was removed. For the mice in the control group, after laparotomy, phosphate buffer without EC 9706 was injected beneath the splenic capsule and spleen was also removed one hour after the injection. Eight hour later, we resected the liver of the nude mice, and expression of E-selectin on vascular endothelial cells of the liver was detected with reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC). Results In the experimental group, 8 hours after injection of EC9706 cells (5×10.6), the results of RT-PCR showed expression of E-selectin mRNA in the liver, and IHC showed a positive protein expression of E-selectin in the cytosol and membrane of hepatic sinus vessels.However, no E-selectin mRNA expression was found in the control group and IHC showed a negative protein expression of E-selectin. Conclusion Human esophageal carcinoma cell line EC9706 can induce balb/c mice liver vascular endothelial cell E-selectin expression, which shows that EC9706 may stay in the liver and form etastatic focus.
ObjectivesTo investigate the expression of E-selectin of adhesive molecule in microvessels in rats′ retina with diabetic retinopathy (DR) at the early, middle, and late phase.MethodsNinety Sprague Dawley (SD) male rats were randomly divided into normal control group (6, 9, and 12 months subgroups with 10 rats in each subgroup) and streptozotocin (STZ) DR group (6, 9, 12 months subgroups with 20 rats in each subgroup). The DR model was set up by one-off celiac injection with STZ at a dose of 60 mg/kg. The rats were executed according to the different time points, and the eyeballs were removed and the digested stretched preparations of retinal microvessels were made. E-selectin of adhesive molecule in microvessels was revealed by EnVision immunohistochemical staining, and the expression of E-selectin in retinal microvessels and the morphological changes were analyzed and observed by Leica-Q550W computer image analytical equipment. The relativity between degree of impairment of DR and expression of E-selectin was detected.ResultsMorphological changes: hyperplasia of endotheliocytes and decreased pericytes were found in 6month DR group; irregularly arranged retinal capillary networks, obviously thickened basilar membrane, many aggregation and adhesion of leucocytes to capillary walls, large areas of acellular capillaries, and opoptosis of endotheliocytes and pericytes were found in 9-and 12-month DR group. Results of quantitative analysis: there was no expression of E-selectin in normal control group, while expression of E-selectin with different degree in each DR group was found. There was significance difference among DR subgroups (Plt;0.01), and the highest expression was found in 6-and 12-month DR group.ConclusionsThe expression of adhesive molecule in microvessels in rats' retina increases as the disease course of DR extends, and has positive correlation with the degree of impairment of microvessels in DR. (Chin J Ocul Fundus Dis, 2005,21:318-321)
【Abstract】Objective To investigate the effects of human interlukin-13 (hIL-13) on the expression of E-selectin and intercellular adhesion molecule-1(ICAM-1) on bovine aortic endothelial cells(BAECs) stimulated by tumor necrosis factor alpha(TNF-α), and to provide experimental basis for hIL-13 inducing immunity endure and relieving the repulsion reaction of xenograft. Methods BAECs were co-cultured with different concentrations of hIL-13 for 2 h and followed by co-cultured with 4 ng/ml TNF-α for 6 h or 18 h. The expressions of E-selectin and ICAM-1 on BAECs were detected by Cell-ELISA. The effect of hIL-13 on activity of BAECs was detected by MTT colorimetry.Results BAECs pretreateded with hIL-13 could inhibit the expression of E-selectin and ICAM-1 induced by TNF-α, and showed a doesdependent manner from 5 ng/ml to 20 ng/ml of hIL-13 (P＜0.01). The experimental result of BAECs activity measured by MTT proved no significant difference in the activities of BAECs in every experimental groups compared with control group’s. Conclusion hIL-13 could inhibit the expression of E-selectin and ICAM-1 on BAECs induced by TNF-α, which may contribute to the xenotransplant immune tolerance.
We in this study measured the site density of E-selectin in order to explore the practical pliability using radionuclide labeling method and γ-imaging of single photon emission computer tomography (SPECT). This method required labeling of antibody with 125I using Indogen method and binding of the labeled antibody to E-selectin. Labeled E-selectin was separated and purified in a Sephadex G25 column. The different fractions of the eluants were imaged, analyzed and quantified with SPECT method. For measuring the saturation curve of E-selectin, 130 μL of E-selectin solution with different concentrations were added in a 48-well plate and incubated overnight at 4℃. After incubation, 130 μL of labeled antibody solution were added and kept incubated for 30 min. The resulted mixture was washed, and the radioactivity in each sample was detected by SPECT. The levels of radioactivity were translated to site densities, and were used to plot a standard curve. The labeled product was quantitatively analyzed with SPECT. The labeling rate of E-selectin was 78%. The saturation curve of different concentration samples showed that when the concentration was in the concentration range of 0-1 mg/mL, the standard curve was y=6 045.7x—51.166, R2=0.997 9. Based on this finding, it could be concluded that γ-imaging is an important tool for analysis of radiolabeled product and determination of site density.
Objective To investigate the effect of CD44 fucosylation on fluid adhesion force of rabbit bone marrow mesenchymal stem cells (BMSCs). Methods The rabbit BMSCs were isolated and purified by density gradient centrifugation combined with adherent culture method. The morphology of cells were observed by inverted microscope, and the cell surface markers of CD44, CD34, CD29, and CD105 were assessed by flow cytometry. BMSCs fucosylated by alpha-(1, 3)-fucosyltransferase Ⅵ (FTⅥ) were as the experimental group, and the non-fucosylated BMSCs were as the control group, and then the positive rate of sialyl-LewisX (sLeX) and the binding rate of E-selectin were detected by flow cytometry. The fucosylated BMSCs resuspended in Hank balanced salt solution (HBSS) were assigned as the experimental group (group A), at same time, the non-fucosylated BMSCs resuspended in HBSS solution as the study control group (group B), and the fucosylated BMSCs resuspended in HBSS solution which was added EDTA as negative control group (group C). The fluid adhesion force of rabbit BMSCs were detected by the parallel flow chamber adhesion test. Results Primary BMSCs mainly shaped as spindle and kept strong growth. The third generation BMSCs were negative for CD34, but positive for CD44, CD29, and CD105. After fucosylation, the positive rate of sLeX in the experimental group was 32.52%±1.76%, which was significantly higher than that in the control group (1.48%±0.51%) (t=29.277, P= 0.000). The binding rate of E-selectin in the experimental group was 41.05%±1.84%, which was also significantly higher than that in the control group (4.33%±0.92%) (t=35.674, P=0.000). With the increase of fluid shear force, the number of BMSCs adhering to the surface of human umbilical vascular endothelial cells (HUVEC) in group A was increased at first and then decreased, while there was few BMSCs adhering to the surface of HUVEC in groups B and C. Under the different fluid shear stress, the number of BMSCs adhered to the surface of HUVEC in group A was significantly higher than that in groups B and C (P<0.05), and there was no significant difference between groups B and C (P>0.05). Conclusion CD44 fucosylation on BMSCs can enhance the fluid adhesion force of rabbit BMSCs.