Objective To observe the protective effect on rat lung by using N-acetyl-L-cysteine(NAC) a inhibiter of nuclear factor-kappa B (NF-κB) in the period of reperfusion. Methods Twenty-four rats were randomly divided into a control group and a trail group.The harvested lung blocks of 12 rats were flushed with and stored in the low-potassium-dextran (LPD) solution at 4℃ for 16 hours. The isolated rat lung reperfusion models were established and the donor lungs were perfused for 1 hour. NAC was used in the trail group but normal saline was used in control group. Partical pressure of oxygen in artery (PaO2), peak airway pressure (PawP) were measured at every 15 min intervals during reperfusion. After reperfusion, the lung tissue wet-to-dry(W/D)ratio, and myeloperoxidase(MPO) activity were obtained. The protein and mRNA expressions of intercellular adhesion molecule-1(ICAM-1), NF-κB were also observed by using immunohistochemistry and semi-quantitative RT-PCR at the end of reperfusion. Results The level of decreased PaO2 and increased PawP in trail group were lower than those in control group at every interval time the sample obtained after reperfusion in 60 min. (Plt;0.01 or lt;0.05). After reperfusion the W/D,MPO, the protein and mRNA expressions of ICAM-1, NF-κB were decreased evidently in trail group than those in control group(Plt;0.01 or lt;0.05). Conclusion Using NAC in the period of reperfusion, can effectively inhibit the expression of NF-κB and ICAM-1,further improve lung respiratory functions.
【Abstract】ObjectiveTo compare the effects of newcastle disease virus (NDV) and adriamycin (ADM) on surface structure and actin of hepatocellular carcinoma cell lines SMMC-7721. Methods SMMC-7721 carcinoma cell lines were divided into 2 groups. NDV was added into one group, while ADM was added into the other group. The cells were then cultured at 5 time phases (8, 16, 24, 36 and 48 h). Intracellular actin and Ca2+ were examined by using immunofluorescence method. CD44 and intercellular adhesion molecule-1 (ICAM-1) were detected by using immunochemical method and flow cytometry, respectively. The change of cellular surface structure was observed by scan electron microscope. Results Cells gradually contracted and turned round over time. It was observed that actin was segmented and cells alignment became disordered. The mean fluorescence intensity of actin decreased in both groups, but it was obvious in NDV group. There were significant differences of fluorescence intensity between 2 groups at the phases of 16 h (P＜0.05), 24 h (P＜0.05), 36 h (P＜0.01) and 48 h (P＜0.05), except the one after 8 h. Intracellular Ca2+ concentration increased gradually in both groups, and the amplifications in NDV group were significantly higher at the phases of 24 h, 36 h and 48 h than those in ADM group (P＜0.01， P＜0.05 and P＜0.01, respectively). There were also differences at 8 and 16 h, but there were no statistical significance. The expression of CD44 in cells decreased. The mean fluorescence intensity of ICAM-1 raised gradually, and then came to peaking at 36 h, but there was no significant difference between two groups. All the above indices between different phases in the same group showed significant differences (P＜0.05). Conclusion Both NDV and ADM could make tumor cells degenerate and rupture, but the effect of NDV is more intensive. It could increase the fragility of cells and hasten the process of cell rupture. Disintegrated cancer cell and changes of adhesion molecule could lead cancer cells be identified, encapsulated, and killed by immune cells under static condition.
Objective To explore the anti-inflammatory effects of ambroxol hydrochloride in chronic obstructive pulmonary disease（COPD）.Methods Thirty Wistar rats were randomly divided into three groups，ie.a control group，a smoking group and an ambroxol group.The rats in the smoking and ambroxol groups were exposed to cigarettes smoking for 12 weeks.Ambroxol hydrochloride was administered via intragastric gavage after 4 weeks smoking in the ambroxol group.After 12 weeks，the expiratory airway resistance(Re) and dynamic lung compliance(CLdyn) were measured.The expression levels of nuclear factor kappa B（NF-κB）and intercellular adhesion molecule-1(ICAM-1) in airway epithelium cell were observed by immunohistochemical method.Results Re was increased and CLdyn was decreased significantly in the smoking and ambroxol groups compared with the control group(all Plt;0.01).Re was lower (Plt;0.01) and CLdyn was higher(Plt;0.05) in the ambroxol group than those in the smoking group.B.The level of NF-κB and ICAM-1 in smoking and ambroxol groups were obviously increased compared with the control group (all Plt;0.05)，which was decreased in the ambroxol group compared with the smoking group(both Plt;0.05).C.The expression of NF-κB was positively correlated with ICAM-1 expression in airway epithelial cells（r=0.924，Plt;0.01）.Conclusions Smoking can increase the airway resistance，reduce the lung compliance and increase the expression of NF-κB and ICAM-1 in airway epithelium.Ambroxol hydrochloride can relieve those effects of smoking，which suggested an anti-inflammatory therapeutic role in COPD.
ObjectiveTo investigate the protective effect of Roux-en-Y gastric bypass surgery on early damage of renal tissue in type 2 diabetes mellitus rats, and explore the mechanism of the protective effects. MethodsDiabetes mellitus animal models were induced by intraperitoneal injection of streptozotocin (STZ, 35 mg /kg) and a high-fat diet.Diabetic rats were divided into three groups randomly (digital table method): diabetes control group (n=8), sham operation group (n=8), and Roux-en-Y gastric bypass group (n=14).Another 8 normal SD rats as the normal control group.The fasting blood glucose, serum total cholesterol (TC), triglyceride (TG), and free fatty acid (FFA) were measured before operation and in 8 weeks after operation; plasma BUN and Cr were measured respectively before operation and in 4 and 8 weeks after operation in each group rats, 24 h urine microalbumin and urine 8-hydroxydeoxyguanosine were measured respectively before operation and in 8 weeks after operation in each group rats.Renal pathological changes were observed and the indexes of kidney hypertrophy, the mean glomerular area (MGA), and the mean glomerular volume (MGV) were analyzed in 8 weeks after operation.The expressions of fibronectin, typeⅣcollagen (CoⅣ), transforming growth factor-β1 (TGF-β1), intercellular adhesion molecule-1(ICAM-1), nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4), and Bcl-2 protein in renal tissues were investigated by immunohistochemical staining. ResultsRoux-en-Y gastric bypass surgery could reduce the blood glucose, blood lipid, MGA, MGV, and the index of kidney hypertrophy of diabetic rats significantly (P < 0.05), improved renal pathological morphology and kidney function (P < 0.05), reduced the protein expressions of fibronectin and CoⅣ, decreased the protein expressions of TGF-β1, ICAM-1, and NOX4, and increased the protein expression of Bcl-2. ConclusionRoux-en-Y gastric bypass surgery can improve kidney function and the pathological damage of diabetes rats, its mechanism may be related to inhibition the protein expressions of TGF-β1, ICAM-1, and NOX4, and increase the protein expression of Bcl-2.
ObjectiveTo study the clinical value of changes of serumα-fetoprotein(AFP) and soluble cell adhesion molecule-1(sICAM-1) levels before and after surgical treatment of primary hepatocellular carcinoma(PHC) as predictors of patient survival. MethodsThe clinical data and followed-up results of 86 patients with hepatocellular carcinoma received hepatectomy or radiofrequency ablation(RFA) in Xijing Hospital and the 451st Hospital of PLA were retrospectivly analyzed. The changes of peripheral blood AFP and sICAM-1 levels in patients before and in 1 month after treatment were observed and all patients were divided into different groups according to the changes in both two markers. Then survival rates of each group were analyzed. ResultsThe patients with AFP < 20μg/L or sICAM-1 < 1 000 U/L before treatment had lower tumor recurrence rate and higher survival rate than patients with elevated serum levels of the both markers(AFP:P=0.018, P < 0.001;sICAM-1:P=0.027, P < 0.001). The larger tumor, late TNM stage, and higher rate of recurrence were associated with elevated serum levels of the both markers(AFP:P=0.016, P=0.026 and P=0.025;sICAM-1:P < 0.001, P=0.024 and P=0.032). The better survival situation was closely related with these cases treated with hepatectomy and their levels of both markers were lower than the above cutoff values both before and after treatment, or leves of both markers above the cut-off values returned to within the normal range after treatment (AFP:P=0.006, P=0.001;sICAM-1:P=0.001, P=0.002). The patients who had simultaneous increase of AFP and sICAM-1 after operation showed the worst tumor-free and overall survivals(P=0.007, P < 0.001). ConclusionTo test the changes of serum AFP and sICAM-1 levels in early stage after treatment for patients who received radical resection of hepatocellular carcinoma has good clinical value for monitoring of tumor recurrence and predict prognosis.
【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.
Objective To investigate the targeted combination and anti-inflammatory effects of anti-intercellular adhesion molecule 1 (ICAM-1) targeted perfluorooctylbromide (PFOB) particles on myocardial ischemia-reperfusion injury in rat model. Methods Seventy-six adult Sprague Dawley rats (male or female, weighing 250-300 g) were selected for experiment. The models of myocardial ischemia-reperfusion injury were established by ligating the left anterior descending coronary artery for 30 minutes in 30 rats. The expression of ICAM-1 protein was detected by immunohistochemistry staining at 6 hours after reperfusion, and the normal myocardium of 10 rats were harvested as control; then the content of interleukin 8 (IL-8) in serum was tested every 6 hours from 6 hours to 48 hours after reperfusion. The other 36 rats were randomly divided into 6 groups (n=6): ischemia-reperfusion injury model/targeted PFOB particles group (group A), ischemia-reperfusion injury model/untargeted PFOB group (group B), normal control/targeted PFOB particles group (group C), normal control/untargeted PFOB particles group (group D), ischemia-reperfusion injury model/normal saline group (group E), and sham operation group (group F). The ischemia-reperfusion injury models were established in groups A, B, and E; while a thread crossed under the coronary artery, which was not ligated after open-chest in group F. After 6 hours of reperfusion, 1 mL of corresponding PFOB particles was injected through juglar vein in groups A, B, C, and D, while 1 mL of nomal saline was injected in group E. Ultrasonography was performed in groups A, B, C, and D before and after injection. The targeted combination was tested by fluorescence microscope. The content of IL-8 was tested after 6 and 24 hours of reperfusion by liquid chip technology in groups A, B, E, and F. Results After 6 hours of reperfusion, the expression of ICAM-1 protein significantly increased in the anterior septum and left ventricular anterior wall of the rat model. The content of IL-8 rised markedly from 6 hours after reperfusion, and reached the peak at 24 hours. Ultrasonography observation showed no specific acoustic enhancement after injection of PFOB particles in groups A, B, C, and D. Targeted combination was observed in the anterior septum and left ventricular anterior wall in group A, but no targeted combination in groups B, C, and D. There was no significant difference in the content of IL-8 among groups A, B, and E after 6 hours of reperfusion (P gt; 0.05), but the content in groups A, B, and E was significantly higher than that in group F (P lt; 0.05). After 24 hours of reperfusion, no sigificant difference was found in the content of IL-8 between groups A and B (P gt; 0.05), but the content of IL-8 in groups A and B were significantly lower than that in group E (P lt; 0.05). Conclusion Anti-ICAM-1 targeted PFOB particles can target to bind and pretect injured myocardium of rat by its anti-inflammation effects.
Objective To study the mechanisms and treatment of ischemia /reperfusion injury, expression of intracellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) were measured, the effect on suppression of ICAM-1 and VCAM-1 by the pyrrolidine dithiocarbamate (PDTC) were investigated. Methods Endothelial cells were divided into 3 groups, hypoxia group: endothelial cells were exposed in hypoxia condition, then returned to reoxygenation condition; the PDTC group: PDTC was added to the endothelial cells in the culture media before exposing to hypoxia condition; control group: endothelial cells underwent treatment. Confocal microscopy was used to detect expression of ICAM-1 and VCAM-1. Results ICAM-1 and VCAM-1 expression were low in endothelial cells of control group, and increased in hypoxia group . ICAM-1 and VCAM-1 expression of endothelial cells in PDTC group werelower than those in hypoxia group , but higher than those in control group. Conclusions It seems that hypoxia/ reoxygenation can activate the endothelial cells and increase the expression of cell adhesion molecules. PDTC can decrease the expression of ICAM-1 and VCAM-1. PDTC may prove benificial in the treatment of ischemia /reperfusion injury.
Abstract: Objective To investigate the effect of modified ultrafiltration on attenuating the inflammatory reaction and endothelial cell activation or damage after cardiopulmonary bypass (CPB). Methods Forty patients undergoing cardiac operation with CPB were randomly divided into two groups. Ult rafiltration group ( n = 20) : patients underwent modified ultrafiltration after CPB; control group ( n = 20 ) : without ultrafiltration. Plasma concentrations of soluble intercellular adhesion molecules-1 ( s ICAM -1) and tumor necrosis factor-α (TNF-α) were determined with enzyme linked immunosorbent assay and radioimmunity pre-operat ively (baseline) , at the end of CPB, 4h and 24h post-operatively in both groups. Results The concentrations of sICAM -1 in the cont rol group at 4h and 24h po st-operatively were higher than those pre-operatively (P lt; 0. 01). The concentrations sICAM -1 in the ultrafiltrat ion group in pre-operatively and at the end of CPB were not significantly different from that of the control group, but they were lower at 4h and 24h post-operatively (269. 6±33. 8Lg/L vs. 409. 6±37. 3Lg/L , 245. 9±32. 2Lg/L vs. 379. 3±35. 7Lg/L ; P lt; 0. 01). In the ultrafiltration group, the concentration of TN F-α at the end of CPB and 4h post-operatively were higher than that pre-operatively (P lt; 0. 01). The concent rations of TNF-α in the ultrafiltration group at 24h post-operatively recoved to the pre-operative level (0. 177±0. 024Lg/L vs. 0. 172±0. 030Lg/L ; P gt;0.05). In the control group, the concentration of TN F-α was higher at the end of CPB than that pre-operatively (P lt;0.01) , and decreased slightly at 4h and 24h post-operatively, but remained higher than those pre-operat ively (0. 264±0.045Lg/L vs. 0.174±0.033Lg/L , 0.218±0.028Lg/L vs. 0.174±0. 033Lg/L ; P lt; 0. 05). Conclus ion CPB is known to induce inflammatory reaction and endothelial cell activation or damage. Modified ultrafiltration appears to attenuate these adverse reactions and is beneficial to postoperative convalescence.