Effects of smoking cessation on expressions of nuclear factor-κB, matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in airway epithelial cells of smoking rats

Objective To investigate the effects of smoking and smoking cessation on airway inflammation and remodeling in chronic obstructive pulmonary diseases through detecting mRNA and protein expressions of nuclear factor-κB (NF-κB), cell matrix metalloproteinase-9 (MMP-9) and cellular tissue inhibitor of metalloproteinase-1 (TIMP-1) in airway epithelial cells of smoking and smoking cessation rats. Methods Twenty-four male Wistar rats were randomly divided into control group, smoking group and smoking cessation group,eight in each group. Hybridization in situ and immunohistochemistry were used to detect mRNA and protein expressions of NF-κB, MMP-9 and TIMP-1 in airway epithelial cells of rats. Results ① Compared with control group (0.29 ± 0.06,0.29±0.06), mRNA and protein expressions of NF-κB in smoking group (0.45±0.04,0.41±0.03) and smoking cessation group (0.40±0.05,0.37±0.03) were higher (all P＜0.05). The mRNA and protein expressions of NF-κB in smoking cessation group were lower than those in smoking group (all P ＜0.05). ②Compared with control group (0.30±0.06,0.30±0.06) ,mRNA and protein expressions of MMP-9 in smoking group (0.52±0.03,0.51±0.07) and smoking cessation group (0.38±0.03,0.33±0.02) were higher (all P＜0.05). The mRNA and protein expressions of MMP-9 in smoking cessation group were lower than those in smoking group (all P＜0.05). ③Compared with control group (0.26±0.04, 0.26±0.04), mRNA and protein expressions of TIMP-1 in smoking group (0.49±0.05,0.37±0.03) and smoking cessation group (0.42±0.04,0.35±0.03) were higher (all P ＜0.05). The mRNA and protein expressions of TIMP-1 in smoking cessation group were lower than those in smoking group (all P ＜ 0.05). ④ Compared with control group (1.00±0.02,1.00±0.02), MMP-9/TIMP-1 mRNA and protein expressions were larger than one in smoking group (1.07±0.14, 1.37±0.19), and less than one in smoking cessation group (0.92±0.13,0.94±0.10) (all P ＜0.05). ⑤The mRNA and protein expressions of NF-κB and MMP-9in each group were positively correlation (r=0.87,0.66,all P ＜0.05). Conclusions In airway epithelial cells of smoking rats, mRNA and protein expressions of NF-κB, MMP-9 and TIMP-1 increase, and MMP-9/TIMP-1 is larger than one. After stoping smoking, mRNA and protein expressions of NF-κB,MMP-9 and TIMP-1 decrease, and MMP-9/TIMP-1 is less than one. This experiment explains that smoking can cause airway inflammation and remodeling, smoking cessation can reduce airway inflammation and remodeling.     

急性冠状动脉综合征患者外周血单个核细胞核因子-κB蛋白表达水平与冠状动脉病变特征的关系

Objective To explore the correlation between expression of nuclear factor kappa B (NF-kappa B) in peripheral blood monouclear cells(PBMC) and the severity of coronary artery lesion in patients with acute coronary syndrome(ACS). Methods The 81 patients were diagnosed with coronary heart disease, and all of them underwent immediate or selective coronary angiography(CAG) The nuclear protein level of activated NF-B was detected by Western blot and semi-quantity image analysis was done. Coronary angiograms were scored according to Gensini integration. The relationship between NF-κB and Gensini score was analyzed. Results The protein expression of NF-κB in PBMC in ACS group was significantly higher than that in stable angina group and control group (0.85±0.18 vs. O.75±0.21 and 0.71±0.23, F=3.72, P<0.05). The protein expression of NF-κB was not correlated with Gensini score(r=0.07, P>0.05). Conclusions The ACS patients have the activation of NF-κB in PBMC. The protein expression of NF-κB is not correlated with Gensini score, which does not suggest an implication of the severity of coronary artery.     

Expression and significance of nuclear factor κB p65 in colon tissues of rats with TNBS-induced colitis

AIM: To investigate the role of NF-κB in the pathogenesis of TNBS-induced colitis in rats. METHODS: Thirty-two healthy adult Sprague-Dawley (SD) rats were randomly divided into four groups of eight each: normal, NS, model Ⅰ, model Ⅱ groups in our study. Rat colitis model was established through 2-,4-,6-trinitrobenzene sulfonic acid (TNBS) enema. At the end of four weeks, the macroscopical and histological changes of the colon were examined and mucosa myeloperoxidase (MPO) activities assayed. NF-κB p65 expression was determined by Western blot assessment in cytoplasmic and nuclear extracts of colon tissue, and the expressions of TNF-α and ICAM-1 protein in colon tissue were examined by immunohistochemistry. The relativities between expression of NF-κB p65 and other parameters were analyzed. RESULTS: TNBS enema resulted in pronounced pathological changes of colonic mucosa in model Ⅱ group (macroscopic and histological injury indices 6.25±1.39 and 6.24±1.04, respectively), which were in accordance with the significantly elevated MPO activity (1.69±0.11). And the nuclear level of NF-κB and expression of TNF-α, ICAM-1 in rats of model II group were higher than that of normal control (9.7±1.96 vs1.7±0.15, 84.09±14.52 vs 16.03±6.21, 77.69±8.09 vs 13.41±4.91 P<0.01), Linear correlation analysis revealed that there were strong correlations between the nuclear level of NF-κB and the tissue positive expression of TNF-α and ICAM-1, MPO activities, macroscopical and histological indices in TNBS-induced colitis, respectively (r= 0.8235, 0.8780, 0.8572, 0.9152, 0.8247; P<0.05). CONCLUSION: NF-κB plays a pivotal role in the pathogenesis of ulcerative colitis, which might account for the up-regulation the expression of TNF-α and ICAM-1.     

Inhibition effect of captopril and losartan on expression of matrix metalloproteinase-1,-9 induced by angiotensin Ⅱ in rat vascular smooth muscle cells

Objective To investigate the effect of angiotensin converting enzyme inhibitor (ACEI) captopril and angiotensin Ⅱ receptor antagonist losartan on the mRNA expression of matrix metalloproteinase (MMP)-1 and MMP-9 in vascular smooth muscle cells induced by angiotensin Ⅱ (Ang Ⅱ ).Methods Male Wistar rats' thoracic aortic vascular smooth muscle cells were cultured in vitro.The cultured cells were divided in to control group,Ang Ⅱ group,captopril group,losartan group,and captopril plus losartan group.Cells in all groups were collected at the culture end-point.MMP-1 and MMP-9 mRNA expressions were detected by RT-PCR method in the collected specimens,and the effects of Ang Ⅱ on MMP-1 and MMP-9 mRNA expression and the intervention effects of captopril and losartan were observed in different Ang Ⅱ concentrations and different action times to vascular smooth muscle cells.Results ( 1 ) MMP-1 mRNA expression gradually increased along with the increments of Ang Ⅱ concentration and the action time (P＜0.05),and the most significant concentration was 10-6 mol/L (P＜0.01).(2)Captopril (5 × 10-6 mol/L) and losartan (5 × 10-6mol/L) inhibited the action of AngⅡ (P＜0.05,P＜0.01).MMP-9 mRNA expression was 0.47±0.03 ,0.86 ± 0.04,0.94±0.14 and 1.12±0.19 vs.0.10±0.04 (P＜0.05,P＜0.01) respectively when Ang Ⅱ concentration was 10-7 ,10-6 ,10-5 and 10-4 mol/L respectively.Captopril (5 × 10-6mol/L) and losartan (5 × 10-6 mol/I) significantly inhibited the MMP-9 mRNA expression which was stimulated by Ang Ⅱ (P＜0.05,P＜0.01),especially in captopril plus losartan group.The MMP-9 mRNA expression increased with the prolonging of stimulating time of Ang Ⅱ,MMP-9 mRNA expression was earlier than that of MMP-1.Conclusions AngⅡ increases the expression of MMP-1 and MMP-9 of vascular smooth muscle cells in a dose-and time-dependence manner.Captopril and losartan inhibit the MMP-1 and MMP-9 mRNA expression of vascular smooth muscle cells induced by Ang Ⅱ ,and the inhibition is the strongest when losartan was combined with captopril.The inhibitive effects is positively correlated to action time.     

Inhibition effect of captopril and losartan on expression of matrix metalloproteinase-1,-9 induced by angiotensin Ⅱ in rat vascular smooth muscle cells

Objective To investigate the effect of angiotensin converting enzyme inhibitor (ACEI) captopril and angiotensin Ⅱ receptor antagonist losartan on the mRNA expression of matrix metalloproteinase (MMP)-1 and MMP-9 in vascular smooth muscle cells induced by angiotensin Ⅱ (Ang Ⅱ ).Methods Male Wistar rats' thoracic aortic vascular smooth muscle cells were cultured in vitro.The cultured cells were divided in to control group,Ang Ⅱ group,captopril group,losartan group,and captopril plus losartan group.Cells in all groups were collected at the culture end-point.MMP-1 and MMP-9 mRNA expressions were detected by RT-PCR method in the collected specimens,and the effects of Ang Ⅱ on MMP-1 and MMP-9 mRNA expression and the intervention effects of captopril and losartan were observed in different Ang Ⅱ concentrations and different action times to vascular smooth muscle cells.Results ( 1 ) MMP-1 mRNA expression gradually increased along with the increments of Ang Ⅱ concentration and the action time (P＜0.05),and the most significant concentration was 10-6 mol/L (P＜0.01).(2)Captopril (5 × 10-6 mol/L) and losartan (5 × 10-6mol/L) inhibited the action of AngⅡ (P＜0.05,P＜0.01).MMP-9 mRNA expression was 0.47±0.03 ,0.86 ± 0.04,0.94±0.14 and 1.12±0.19 vs.0.10±0.04 (P＜0.05,P＜0.01) respectively when Ang Ⅱ concentration was 10-7 ,10-6 ,10-5 and 10-4 mol/L respectively.Captopril (5 × 10-6mol/L) and losartan (5 × 10-6 mol/I) significantly inhibited the MMP-9 mRNA expression which was stimulated by Ang Ⅱ (P＜0.05,P＜0.01),especially in captopril plus losartan group.The MMP-9 mRNA expression increased with the prolonging of stimulating time of Ang Ⅱ,MMP-9 mRNA expression was earlier than that of MMP-1.Conclusions AngⅡ increases the expression of MMP-1 and MMP-9 of vascular smooth muscle cells in a dose-and time-dependence manner.Captopril and losartan inhibit the MMP-1 and MMP-9 mRNA expression of vascular smooth muscle cells induced by Ang Ⅱ ,and the inhibition is the strongest when losartan was combined with captopril.The inhibitive effects is positively correlated to action time.     

Inhibition effect of captopril and losartan on expression of matrix metalloproteinase-1,-9 induced by angiotensin Ⅱ in rat vascular smooth muscle cells

Objective To investigate the effect of angiotensin converting enzyme inhibitor (ACEI) captopril and angiotensin Ⅱ receptor antagonist losartan on the mRNA expression of matrix metalloproteinase (MMP)-1 and MMP-9 in vascular smooth muscle cells induced by angiotensin Ⅱ (Ang Ⅱ ).Methods Male Wistar rats' thoracic aortic vascular smooth muscle cells were cultured in vitro.The cultured cells were divided in to control group,Ang Ⅱ group,captopril group,losartan group,and captopril plus losartan group.Cells in all groups were collected at the culture end-point.MMP-1 and MMP-9 mRNA expressions were detected by RT-PCR method in the collected specimens,and the effects of Ang Ⅱ on MMP-1 and MMP-9 mRNA expression and the intervention effects of captopril and losartan were observed in different Ang Ⅱ concentrations and different action times to vascular smooth muscle cells.Results ( 1 ) MMP-1 mRNA expression gradually increased along with the increments of Ang Ⅱ concentration and the action time (P＜0.05),and the most significant concentration was 10-6 mol/L (P＜0.01).(2)Captopril (5 × 10-6 mol/L) and losartan (5 × 10-6mol/L) inhibited the action of AngⅡ (P＜0.05,P＜0.01).MMP-9 mRNA expression was 0.47±0.03 ,0.86 ± 0.04,0.94±0.14 and 1.12±0.19 vs.0.10±0.04 (P＜0.05,P＜0.01) respectively when Ang Ⅱ concentration was 10-7 ,10-6 ,10-5 and 10-4 mol/L respectively.Captopril (5 × 10-6mol/L) and losartan (5 × 10-6 mol/I) significantly inhibited the MMP-9 mRNA expression which was stimulated by Ang Ⅱ (P＜0.05,P＜0.01),especially in captopril plus losartan group.The MMP-9 mRNA expression increased with the prolonging of stimulating time of Ang Ⅱ,MMP-9 mRNA expression was earlier than that of MMP-1.Conclusions AngⅡ increases the expression of MMP-1 and MMP-9 of vascular smooth muscle cells in a dose-and time-dependence manner.Captopril and losartan inhibit the MMP-1 and MMP-9 mRNA expression of vascular smooth muscle cells induced by Ang Ⅱ ,and the inhibition is the strongest when losartan was combined with captopril.The inhibitive effects is positively correlated to action time.     

Hepatic preconditioning of doxorubicin in stop-flow chemotherapy:NF-κB/IκB-α pathway and expression of HSP72

AIM: To provide hepatic protection through administration of doxorubicin before stop-flow chemotherapy (SFC) and to investigate the expression of heat shock protein 72 (HSP72) and role of nuclear factor kappa B (NF-κB) in this effect. METHODS: The hepatic preconditioning of doxorubicin was established in a porcine model by injection of doxorubicin (1 mg/kg) before SFC. The experimental animals were randomized into two groups: groups receiving doxorubicin (DOX) and normal saline (NS). Serial serum and tissue samples were taken from both groups to evaluate the protection of doxorubicin. Western blot and immunoprecipitation were applied to detect the expression of HSP72, NF-κB p65 protein, inhibitor κB-α (IκB-α) and phosphorylated IκB-α as well. The expression of tumor necrosis factor α (TNF-α) was estimated by semiquantitative RT-PCR. And the extent of the hepatic injury was estimated with the level of serum aminotransferases. RESULTS: An abundance production of HSP72 in porcine liver was observed after 24 h of intravenous administration of doxorubicin, but without any change in the expression of NF-κB p65 subunit in cytoplasm. NF-κB p65 subunit accumulated in nuclei at the end of SFC and reached its highest level at 30 min after the restoration of the abdominal circulation and decreased gradually during the 6 h after SFC in NS group, while there was little change in DOX group. There was also a slight decrease of IκB-α at 30 min after the restoration of the abdominal circulation in NS group accompanying with the appearance of phosphorylated IκB-α. The expression of TNF-α was significantly higher in NS group than that in DOX group (average 1.40±0.17 vs 0.62±0.22, P<0.01) at serial time points after SFC. Serum ALT and AST levels of NS group were higher after 24 h than those of DOX group (93.2±7.8 IU/L vs 53.3±13.9 IU/L, 217.0±29.4 IU/L vs 155.0±15.6 IU/L for ALT and AST respectively, P<0.05) and after 48 h than those of DOX group (66.6±18.1 IU/L vs 43.3±16.7 IU/L, 174.4±21.3 IU/L vs 125.7±10.5 IU/L for ALT and AST respectively, P<0.05). CONCLUSION: Doxorubicin renders the liver to be tolerant to the hepatic influence in SFC in a porcine model through the NF-κB/IκB-αpathway with the expression of HSP72.     

Nuclear factor-KB decoy oligodeoxynucleotides attenuates ischemia/reperfusion injury in rat liver graft

AIM: To evaluate the protective effect of NF-κB decoy oligodeoxynucleotides (ODNs) on ischemia/reperfusion (I/R) injury in rat liver graft. METHODS: Orthotopic syngeneic rat liver transplantation was performed with 3 h of cold preservation of liver graft in University of Wisconsin solution containing phosphorothioated double-stranded NF-κB decoy ODNs or scrambled ODNs. NF-κB decoy ODNs or scrambled ODNs were injected intravenously into donor and recipient rats 6 and 1 h before operation, respectively. Recipients were killed 0 to 16 h after liver graft reperfusion. NF-κB activity in the liver graft was analyzed by electrophoretic mobility shift assay (EMSA). Hepatic mRNA expression of TNF-α, IFN-γ and intercellular adhesion molecule-1 (ICAM-1) were determined by semiquantitative RT-PCR. Serum levels of TNF-α and IFN-γ were measured by enzyme-linked immunosorbent assays (ELISA). Serum level of alanine transaminase (ALT) was measured using a diagnostic kit. Liver graft myeloperoxidase (MPO) content was assessed. RESULTS: NF-κB activation in liver graft was induced in a time-dependent manner, and NF-κB remained activated for 16 h after graft reperfusion. NF-κB activation in liver graft was significant at 2 to 8 h and slightly decreased at 16 h after graft reperfusion. Administration of NF-κB decoy ODNs significantly suppressed NF-κB activation as well as mRNA expression of TNF-α, IFN-γ and ICAM-1 in the liver graft. The hepatic NF-κB DNA binding activity [presented as integral optical density (IOD) value] in the NF-κB decoy ODNs treatment group rat was significantly lower than that of the I/R group rat (2.16±0.78 vs 36.78 ±6.35 and 3.06±0.84 vs 47.62± 8.71 for IOD value after 4 and 8 h of reperfusion, respectively, P<0.001). The hepatic mRNA expression level of TNF-α, IFN-γ and ICAM-1 [presented as percent of p-actin mRNA (%)] in the NF-κB decoy ODNs treatment group rat was significantly lower than that of the I/R group rat (8.31 ±3.48 vs 46.37±10.65 and 7.46± 3.72 vs 74.82±12.25 for hepatic TNF-a mRNA, 5.58±2.16 vs 50.46±9.35 and 6.47±2.53 vs 69.72±13.41 for hepatic IFN-y mRNA, 6.79 ±2.83 vs 46.23±8.74 and 5.28±2.46 vs 67.44±10.12 for hepatic ICAM-1 mRNA expression after 4 and 8 h of reperfusion, respectively, P<0.001). Administration of NF-κB decoy ODNs almost completely abolished the increase of serum level of TNF-α and IFN-γ induced by hepatic ischemia/reperfusion, the serum level (pg/mL) of TNF-α and IFN-γ in the NF-kB decoy ODNs treatment group rat was significantly lower than that of the I/R group rat (42.7±13.6 vs 176.7±15.8 and 48.4±15.1 vs 216.8±17.6 for TNF-α level, 31.5±12.1 vs 102.1±14.5 and 40.2±13.5 vs 118.6±16.7 for IFN-γ level after 4 and 8 h of reperfusion, respectively, P<0.001). Liver graft neutrophil recruitment indicated by MPO content and hepatocellular injury indicated by serum ALT level were significantly reduced by NF-κB decoy ODNs, the hepatic MPO content (A655) and serum ALT level (IU/L) in the NF-κB decoy ODNs treatment group rat was significantly lower than that of the I/R group rat (0.17±0.07 vs 1.12 ±0.25 and 0.46±0.17 vs 1.46±0.32 for hepatic MPO content, 71.7±33.2 vs 286.1±49.6 and 84.3±39.7 vs 467.8±62.3 for ALT level after 4 and 8 h of reperfusion, respectively, P< 0.001). CONCLUSION: The data suggest that NF-κB decoy ODNs protects against I/R injury in liver graft by suppressing NF-κB activation and subsequent expression of proinflammatory mediators.     

Nuclear factor-kappaB activation on the reactive oxygen species in acute necrotizing pancreatitic rats

AIM: To investigate the potential role of nuclear factor kappa-B (NF-KB) activation on the reactive oxygen species in rat acute necrotizing pancreatitis (ANP) and to assess the effect of pyrrolidine dithiocarbamate (PDTC, an inhibitor of NF-KB). METHODS: Rat ANP model was established by retrograde injection of 5% sodium taurocholate into biliopancreatic duct. Rats were randomly assigned to three groups (10 rats each): Control group, ANP group and PDTC group. At the 6th h of the model, the changes of the serum amylase, nitric oxide (NO), malondialdehyde (MDA), superoxide dismutase (SOD) and pancreatic morphological damage were observed. The expressions of inducible nitric oxide (iNOS) were observed by SP immunohistochemistry. And the expressions of NF-κB p65 subunit mRNA were observed by hybridization in situ. RESULTS: Serum amylase and NO level decreased significantly in ANP group as compared with PDTC administrated group [(7170.40±1308.63) U/L vs(4 074.10±1719.78) U/L, P<0.05], [(76.95±9.04) μmol/L vs(65.18±9.02)μmol/L, P<0.05] respectively. MDA in both ANP and PDTC group rose significantly over that in control group [(9.88±1.52) nmol/L, (8.60±1.41) nmol/L, vs(6.04±1.78) nmol/L, P<0.05], while there was no significant difference between them. SOD levels in both ANP and PDTC group underwent a significant decrease as compared with that in control t(3 214.59±297.74) NU/mL, (3 260.62±229.44) NU/mL, vs(3 977.80±309.09) NU/mL, P<0.05], but there was no significant difference between them. Though they were still higher than those in Control group, pancreas destruction was slighter in PDTC group, iNOS expression and NF-κB p65 subunit mRNA expression were lower in PDTC group as compared with ANP group. CONCLUSION: We conclude that correlation among NF-KB activation, serum amylase, reactive oxygen species level and tissue damage suggests a key role of NF-κB in the pathogenesis of ANP. Inhibition of NF-κB activation may reverse the pancreatic damage of rat ANP and the production of reactive oxygen species.     

Effect of tumor necrosis factor-α with different levels of iodine on expression of Na+/I- symporter in cultured lactating mammary cells

Objective To observe the expression of Na+/I- symporter(NIS) in cultured lactating mammary cells with different levels of iodine and the effect of tumor necrosis factor-α(TNF-α). Methods Original generation of mouse lactating mammary cells cultured in vitro were divided into low iodine group Ⅰ (LI-Ⅰ), low iodine group Ⅱ (LI-Ⅱ), adequate iodine group(AI), high iodine group Ⅰ(HI-Ⅰ), and high iodine group Ⅱ(HI-Ⅱ). Cells were cultured in DEME/F12 culture medium for 24 h with different concentrations of iodine (0,5,50,3000 and 10 000 μg/L, respectively), and TNF-α( 10-2 mg/L) was added to some of cultured cells for 24 h. The expression of NIS mRNA of lactating mammary cells was determined by real-time quantitative PCR and the expression of NIS protein was detected by In-Cell Western. Results In iodine alone group, the expression of NIS mRNA in LI-Ⅰ group [ (64.66 ± 14.99) x 10-4] was higher than that of AI group[ (22.76 ± 7.36) × 10-4, P ＜ 0.01 ]; HI-I group[ (10.18 ±3.53) × 10-4] and HI-Ⅱ group[ (8.59 ± 2.89) × 10-4] were lower than that of AI group(all P ＜ .0.05); With increased iodine concentration, the expression of NIS mRNA decreased. The expression of NIS mRNA in LI-Ⅰ group [(2.72 ± 0.45) × 10-4], LI-Ⅱ group[ (2.69 ± 0.68) × 10-4] and AI group[(1.80 ± 0.67) × 10-4] with iodine plus TNF-o were all lower than that of LI-Ⅰ group, LI-Ⅱ group[ (29.82 ± 4.47 ) × 10-4], and AI group without TNF-α (all P ＜ 0.01). In iodine plus TNF-α, the expression of NIS mRNA in HI-Ⅰ group[(6.58 ± 2.87) × 10-4] and HI-Ⅱ[(7.04 ± 1.36) × 10-4] group were all higher than that of AI group(all P ＜ 0.05); With increased iodine deficiency or iodine excess, the expression of NIS mRNA increased. With increased iodine concentration, the expression of NIS protein decreased in iodine alone group. The expression of NIS protein in iodine plus TNF-α was all lower than that in iodine alone group. In iodine plus TNF-α, the expression of NIS protein increased in both iodine deficiency and iodine excess conditions. Conclusions Iodine may decrease the expression of NIS mRNA and protein of lactating mammary cells. The expression of NIS mRNA and protein of lactating mammary cells was inhibited by TNF-α under different levels of iodine.     

Effects of hypoxia, hyperoxia on the regulation of expression and activity of matrix metalloproteinase-2 in hepatic stellate cells

AIM To study the effects of hypoxia,hyperoxia on theregulation of expression and activity of matrixmetalloproteinase-2 (MMP-2) in hepatic stellate cells(HSC).METHODS The expressions of MMP-2,tissue inhibitor ofmatrix metalloproteinass-2 (TIMP-2) and membrane typematrix matalloproteinass-1 (MT1-MMP) in cultured rat HSCwere detected by immunocytochemistry (ICC) and in situhybridization (ISH).The contents of MMP-2 and TIMP-2 inculture supernatant were detected with ELISA and theactivity of MMP-2 in supernatant was revealed byzymography.RESULTS In the situation of hypoxia for 12h,theexpression of MMP-2 protein was enhanced (hypoxiagroup positive indexes:5.7±2.0,n=10;control:3.2±1.0,n=7;P<0.05),while TIMP-2 protein was decreasedin HSC (hypoxla group positive indexes:2.5±0.7,n=10;control:3.6±1.0,n=7;P<0.05),and the activity(total A) of MMP-2 in suparnatant declined obviously(hypoxla group:7.334±1.922,n=9;control:17.277±7.424,n=11;P<0.01).Compared the varied duration ofhypoxia,the changes of expressions Including mRNA andprotein level as well as activity of MMP-2 were mostnotable in 6 h group.The highest value (Ahypoxla~-Acontrol) ofthe protein and the most intense signal of mRNA were inthe period of hypoxia for 6 h,along with the lowestactivity of MMP-2.In the situation of hyparoxia for 12 h,the contents (A_(450)) of MMP-2 and TIMP-2 in supernatantwere both higher than those in the control,especially theTIMP-2 (hyperoxla group:0.0499±0.0144,n=16;control:0.0219±0.0098,n=14;P<0.01),and so wasthe activity of MMP-2 (hyperoxia group:5.252±0.771,n=14;control:4.304±1.083,n=12;P<0.05),and the expression of MT1-MMP was increased.CONCLUSION HSC is sensitive to the oxygen,hypoxiaenhances the expression of MMP-2 and the effect is moremarked at the early stage;hyperoxia mainly raises theactivity of MMP-2.     

Effect of resveratrol on activation of nuclear factor kappa-B and inflammatory factors in rat model of acute pancreatitis

AIM: To observe the effect of resveratrol on nuclear factor Kappa-B (NF-κB) activation and the inflammatory response in sodium taurocholate-induced pancreatitis in rats. METHODS: Seventy-two male SD rats were randomly divided into three groups: sham operation group (control), severe acute pancreatitis (SAP) group, and severe acute pancreatitis group treated with resveratrol (RES). A SAP model was established by injecting 4% sodium taurocholate 1 mL/kg through puncturing the pancreatic duct. In Res group, Res was given at 30 mg/kg b.m. intraperitoneally after the SAP model was successfully established. Eight animals from each group were sacrificed at 3, 6 and 12 h after modeling. The expression of NF-κB activation of pancreas was detected by irnmunohistochemical staining, whereas the levels of TNF-α and IL-8 in pancreatic tissues were estimated by radioimrnunoassay. The pathological changes of pancreas and lungs were examined microscopically. RESULTS: Much less hyperemia, edema, dust-colored necrotic focus and soaps were noticed in pancreas in RES group than in SAP group. In RES group, hemorrhage, exudates and infiltration of inflammatory cells in pancreas and interstitial edema, destruction of alveolar wall in lung were significantly less than in SAP group. In the SAP group, the activation of NF-κB in pancreatic tissues was enhanced significantly at any measure point compared with control group (64.23±10.72% vs 2.56±0.65%, 55.86±11.34% vs 2.32±0.42%, 36.23±2.30% vs 2.40±0.36%,P<0.01), TNF-α, IL-8 were also increased and reached their peak at 6 h and then declined. The activation of NF-κB and the levels of TNF-α and IL-8 in RES group were significantly lower than those in SAP group (P<0.01): activation (52.63±9.45% vs 64.23±10.72%, 40.52±8.40% vs 55.86±11.34%, 29.83±5.37% vs 36.23±2.30%), TNF-α (132.76±15.68 pg/mL vs 158.36±12.58 pg/mL, 220.32±23.57 pg/mL vs 247.67± 11.62 pg/mL, 175.68±18.43 pg/mL vs 197.35±12.57 pg/mL) and IL-8 (0.62±0.21 μg/L vs 0.83±0.10 μg/L, 1.10±0.124 μg/L vs1.32±0.18 μg/L, 0.98±0.16 μg/L vs 1.27±0.23μg/L). CONCLUSION: The activation of NF-KB is involved in the inflammatory response of rats with SAP. Resveratrol could effectively inhibit the expression of NF-κB activation, alleviate the severity of SAP through its anti-inflammatory effects and regulate the inflammatory mediators.     

Expression and significance of nuclear factor κB p65 in colon tissues of rats with TNBS-induced colitis

AIM: To investigate the role of NF-κB in the pathogenesis of TNBS-induced colitis in rats. METHODS: Thirty-two healthy adult Sprague-Dawley (SD) rats were randomly divided into four groups of eight each: normal, NS, model Ⅰ, model Ⅱ groups in our study. Rat colitis model was established through 2-,4-,6-trinitrobenzene sulfonic acid (TNBS) enema. At the end of four weeks, the macroscopical and histological changes of the colon were examined and mucosa myeloperoxidase (MPO)activities assayed. NF-~B p65 expression was determined by Western blot assessment in cytoplasmic and nuclear extracts of colon tissue, and the expressions of TNF-α (and ICAM-1 protein in colon tissue were examined by immunohistochemistry. The relativities between expression of NF-κB p65 and other parameters were analyzed. RESULTS: TNBS enema resulted in pronounced pathological changes of colonic mucosa in model Ⅱ group (macroscopic and histological injury indices 6.25&#177;1.39 and 6.24&#177;1.04, respectively), which were in accordance with the significantly elevated MPO activity (1.69+0.11). And the nuclear level of NF-κB and expression of TNF-α, ICAM-1 in rats of model Ⅱ group were higher than that of normal control(9.7&#177;1.96 vs 1.7&#177;0.15, 84.09&#177;14.52 vs 16.03&#177;6.21,77.69&#177;8.09 vs 13.41&#177;4.91 P&lt;0.01), Linear correlation analysis revealed that there were strong correlations between the nuclear level of NF-κB and the tissue positive expression of TNF-α and ICAM-1, MPO activities, macroscopical and histological indices in TNBS-induced colitis, respectively (r = 0.8235, 0.8780, 0.8572, 0.9152,0.8247; P&lt;0.05). CONCLUSION: NF-κB plays a pivotal role in the pathogenesis of ulcerative colitis, which might account for the up-regulation the expression of TNF-α and ICAM-1.     

不同亚型卒中患者血清基质金属蛋白酶-2含量及其意义

Objective To investigate the change of serum matix metalloproteinase-2(MMP-2) level and its significance in patients with acute ischemic stroke of different subtypes. Methods Seventy-seven patients with acute ischemic stroke were classified into large-artery atherosclerosis (LAA) (n =29, 37. 66% ), small artery occlusion (SAO, lacunar infarction) (n =23, 29.87%), cardioembolism (CE) (n = 13,16. 88%), stroke of undemonstrated etiology (SUE) (n = 7, 9.09% ), and stroke of other demonstrated etiology (SOE) (n = 5, 6. 49%) according to the TOAST criteria. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum MMP-2 in patients with acute ischemic stroke at 24 hours and 7 days, and they were compared with 42 controls. Results The serum MMP-2 levels at 24 hours and 7 days of the onset of symptoms in the acute ischemic stroke group according to the TOAST criteria were 189. 55 ±24.79 and 307.46 ±84. 16 ng/ml respectively, and they were all significantly higher than 159.76 ± 10. 32 ng/ml in the control group (all P <0.05). Among all the TOAST subtypes, SOE and SUE were not analyzed because of the small numbers of cases; among other subtypes, the serum MMP-2 levels at 24 hours of the onset of symptoms in the LAA, SAO and CE groups were 218. 60 ± 13.42,175.21 ±9.92, and 167.26 ±9.7 ng/ml respectively, and they were all significantly higher than those in the control group (all P < 0. 05); at day 7 of the onset of symptoms they were 404.75 ± 10. 30, 293.18 ± 10.91, and 211.81 ±11.14 ng/ml respectively, and they were also significantly higher than those in the control group (all P < 0.05). Among those, the LAA group was increased significantly (P < 0. 01). Conclusions The serum MMP-2 levels were increased in patients with acute cerebral infarction. "l'ne changes of the serum MMP-2 levels in each TOAST subtype group were different. The LAA group increased most significantly, which supported the different views of the etiology of cerebral infarction subtypes. The serum MMP-2 plays an important role in the process of cerebral infarction of the LAA type.     

不同亚型卒中患者血清基质金属蛋白酶-2含量及其意义

Objective To investigate the change of serum matix metalloproteinase-2(MMP-2) level and its significance in patients with acute ischemic stroke of different subtypes. Methods Seventy-seven patients with acute ischemic stroke were classified into large-artery atherosclerosis (LAA) (n =29, 37. 66% ), small artery occlusion (SAO, lacunar infarction) (n =23, 29.87%), cardioembolism (CE) (n = 13,16. 88%), stroke of undemonstrated etiology (SUE) (n = 7, 9.09% ), and stroke of other demonstrated etiology (SOE) (n = 5, 6. 49%) according to the TOAST criteria. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum MMP-2 in patients with acute ischemic stroke at 24 hours and 7 days, and they were compared with 42 controls. Results The serum MMP-2 levels at 24 hours and 7 days of the onset of symptoms in the acute ischemic stroke group according to the TOAST criteria were 189. 55 ±24.79 and 307.46 ±84. 16 ng/ml respectively, and they were all significantly higher than 159.76 ± 10. 32 ng/ml in the control group (all P <0.05). Among all the TOAST subtypes, SOE and SUE were not analyzed because of the small numbers of cases; among other subtypes, the serum MMP-2 levels at 24 hours of the onset of symptoms in the LAA, SAO and CE groups were 218. 60 ± 13.42,175.21 ±9.92, and 167.26 ±9.7 ng/ml respectively, and they were all significantly higher than those in the control group (all P < 0. 05); at day 7 of the onset of symptoms they were 404.75 ± 10. 30, 293.18 ± 10.91, and 211.81 ±11.14 ng/ml respectively, and they were also significantly higher than those in the control group (all P < 0.05). Among those, the LAA group was increased significantly (P < 0. 01). Conclusions The serum MMP-2 levels were increased in patients with acute cerebral infarction. "l'ne changes of the serum MMP-2 levels in each TOAST subtype group were different. The LAA group increased most significantly, which supported the different views of the etiology of cerebral infarction subtypes. The serum MMP-2 plays an important role in the process of cerebral infarction of the LAA type.     

不同亚型卒中患者血清基质金属蛋白酶-2含量及其意义

Objective To investigate the change of serum matix metalloproteinase-2(MMP-2) level and its significance in patients with acute ischemic stroke of different subtypes. Methods Seventy-seven patients with acute ischemic stroke were classified into large-artery atherosclerosis (LAA) (n =29, 37. 66% ), small artery occlusion (SAO, lacunar infarction) (n =23, 29.87%), cardioembolism (CE) (n = 13,16. 88%), stroke of undemonstrated etiology (SUE) (n = 7, 9.09% ), and stroke of other demonstrated etiology (SOE) (n = 5, 6. 49%) according to the TOAST criteria. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum MMP-2 in patients with acute ischemic stroke at 24 hours and 7 days, and they were compared with 42 controls. Results The serum MMP-2 levels at 24 hours and 7 days of the onset of symptoms in the acute ischemic stroke group according to the TOAST criteria were 189. 55 ±24.79 and 307.46 ±84. 16 ng/ml respectively, and they were all significantly higher than 159.76 ± 10. 32 ng/ml in the control group (all P <0.05). Among all the TOAST subtypes, SOE and SUE were not analyzed because of the small numbers of cases; among other subtypes, the serum MMP-2 levels at 24 hours of the onset of symptoms in the LAA, SAO and CE groups were 218. 60 ± 13.42,175.21 ±9.92, and 167.26 ±9.7 ng/ml respectively, and they were all significantly higher than those in the control group (all P < 0. 05); at day 7 of the onset of symptoms they were 404.75 ± 10. 30, 293.18 ± 10.91, and 211.81 ±11.14 ng/ml respectively, and they were also significantly higher than those in the control group (all P < 0.05). Among those, the LAA group was increased significantly (P < 0. 01). Conclusions The serum MMP-2 levels were increased in patients with acute cerebral infarction. "l'ne changes of the serum MMP-2 levels in each TOAST subtype group were different. The LAA group increased most significantly, which supported the different views of the etiology of cerebral infarction subtypes. The serum MMP-2 plays an important role in the process of cerebral infarction of the LAA type.     

不同亚型卒中患者血清基质金属蛋白酶-2含量及其意义

Objective To investigate the change of serum matix metalloproteinase-2(MMP-2) level and its significance in patients with acute ischemic stroke of different subtypes. Methods Seventy-seven patients with acute ischemic stroke were classified into large-artery atherosclerosis (LAA) (n =29, 37. 66% ), small artery occlusion (SAO, lacunar infarction) (n =23, 29.87%), cardioembolism (CE) (n = 13,16. 88%), stroke of undemonstrated etiology (SUE) (n = 7, 9.09% ), and stroke of other demonstrated etiology (SOE) (n = 5, 6. 49%) according to the TOAST criteria. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum MMP-2 in patients with acute ischemic stroke at 24 hours and 7 days, and they were compared with 42 controls. Results The serum MMP-2 levels at 24 hours and 7 days of the onset of symptoms in the acute ischemic stroke group according to the TOAST criteria were 189. 55 ±24.79 and 307.46 ±84. 16 ng/ml respectively, and they were all significantly higher than 159.76 ± 10. 32 ng/ml in the control group (all P <0.05). Among all the TOAST subtypes, SOE and SUE were not analyzed because of the small numbers of cases; among other subtypes, the serum MMP-2 levels at 24 hours of the onset of symptoms in the LAA, SAO and CE groups were 218. 60 ± 13.42,175.21 ±9.92, and 167.26 ±9.7 ng/ml respectively, and they were all significantly higher than those in the control group (all P < 0. 05); at day 7 of the onset of symptoms they were 404.75 ± 10. 30, 293.18 ± 10.91, and 211.81 ±11.14 ng/ml respectively, and they were also significantly higher than those in the control group (all P < 0.05). Among those, the LAA group was increased significantly (P < 0. 01). Conclusions The serum MMP-2 levels were increased in patients with acute cerebral infarction. "l'ne changes of the serum MMP-2 levels in each TOAST subtype group were different. The LAA group increased most significantly, which supported the different views of the etiology of cerebral infarction subtypes. The serum MMP-2 plays an important role in the process of cerebral infarction of the LAA type.     

不同亚型卒中患者血清基质金属蛋白酶-2含量及其意义

Objective To investigate the change of serum matix metalloproteinase-2(MMP-2) level and its significance in patients with acute ischemic stroke of different subtypes. Methods Seventy-seven patients with acute ischemic stroke were classified into large-artery atherosclerosis (LAA) (n =29, 37. 66% ), small artery occlusion (SAO, lacunar infarction) (n =23, 29.87%), cardioembolism (CE) (n = 13,16. 88%), stroke of undemonstrated etiology (SUE) (n = 7, 9.09% ), and stroke of other demonstrated etiology (SOE) (n = 5, 6. 49%) according to the TOAST criteria. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum MMP-2 in patients with acute ischemic stroke at 24 hours and 7 days, and they were compared with 42 controls. Results The serum MMP-2 levels at 24 hours and 7 days of the onset of symptoms in the acute ischemic stroke group according to the TOAST criteria were 189. 55 ±24.79 and 307.46 ±84. 16 ng/ml respectively, and they were all significantly higher than 159.76 ± 10. 32 ng/ml in the control group (all P <0.05). Among all the TOAST subtypes, SOE and SUE were not analyzed because of the small numbers of cases; among other subtypes, the serum MMP-2 levels at 24 hours of the onset of symptoms in the LAA, SAO and CE groups were 218. 60 ± 13.42,175.21 ±9.92, and 167.26 ±9.7 ng/ml respectively, and they were all significantly higher than those in the control group (all P < 0. 05); at day 7 of the onset of symptoms they were 404.75 ± 10. 30, 293.18 ± 10.91, and 211.81 ±11.14 ng/ml respectively, and they were also significantly higher than those in the control group (all P < 0.05). Among those, the LAA group was increased significantly (P < 0. 01). Conclusions The serum MMP-2 levels were increased in patients with acute cerebral infarction. "l'ne changes of the serum MMP-2 levels in each TOAST subtype group were different. The LAA group increased most significantly, which supported the different views of the etiology of cerebral infarction subtypes. The serum MMP-2 plays an important role in the process of cerebral infarction of the LAA type.     

不同亚型卒中患者血清基质金属蛋白酶-2含量及其意义

Objective To investigate the change of serum matix metalloproteinase-2(MMP-2) level and its significance in patients with acute ischemic stroke of different subtypes. Methods Seventy-seven patients with acute ischemic stroke were classified into large-artery atherosclerosis (LAA) (n =29, 37. 66% ), small artery occlusion (SAO, lacunar infarction) (n =23, 29.87%), cardioembolism (CE) (n = 13,16. 88%), stroke of undemonstrated etiology (SUE) (n = 7, 9.09% ), and stroke of other demonstrated etiology (SOE) (n = 5, 6. 49%) according to the TOAST criteria. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum MMP-2 in patients with acute ischemic stroke at 24 hours and 7 days, and they were compared with 42 controls. Results The serum MMP-2 levels at 24 hours and 7 days of the onset of symptoms in the acute ischemic stroke group according to the TOAST criteria were 189. 55 ±24.79 and 307.46 ±84. 16 ng/ml respectively, and they were all significantly higher than 159.76 ± 10. 32 ng/ml in the control group (all P <0.05). Among all the TOAST subtypes, SOE and SUE were not analyzed because of the small numbers of cases; among other subtypes, the serum MMP-2 levels at 24 hours of the onset of symptoms in the LAA, SAO and CE groups were 218. 60 ± 13.42,175.21 ±9.92, and 167.26 ±9.7 ng/ml respectively, and they were all significantly higher than those in the control group (all P < 0. 05); at day 7 of the onset of symptoms they were 404.75 ± 10. 30, 293.18 ± 10.91, and 211.81 ±11.14 ng/ml respectively, and they were also significantly higher than those in the control group (all P < 0.05). Among those, the LAA group was increased significantly (P < 0. 01). Conclusions The serum MMP-2 levels were increased in patients with acute cerebral infarction. "l'ne changes of the serum MMP-2 levels in each TOAST subtype group were different. The LAA group increased most significantly, which supported the different views of the etiology of cerebral infarction subtypes. The serum MMP-2 plays an important role in the process of cerebral infarction of the LAA type.