2013, 21(07):583-588.
Abstract:Aim To explore the effect of momordicin on nuclear translocation of inflammatory factor nuclear factor-kappa B (NF-κB) in ApoE-/- mice and further analyze the molecular mechanisms related to momordicin anti-inflammatory. Methods 40 male ApoE-/- mice, 6 weeks old, were randomly divided into four groups: normal food group, high fat high cholesterol group, high fat high cholesterol and momordicin group, normal food and momordicin group. After feeding 12 weeks, all mice were removed eyeball in order to obtain blood preparation. Level of inflammatory factors (IL-1β, TNF-α, IL-6, IFN-γ) and anti-inflammatory factors (IL-10) were measured by ELISA, IκB expression of aorta was analyzed by immunohistochemistry. Gene and protein expression was analyzed by semi-quantitative RT-PCR and Western blot, respectively. Results After fed with high fat high cholesterol about 12 weeks, inflammatory factors (IL-1β, TNF-α, IL-6, IFN-γ) level increased compared with normal food group(P<0.01, n5), but the level of IL-10 can not be up-regulated. After momordicin treating, inflammatory factors level decreased, yet IL-10 can not increase. Not only momordicin inhibited p65 mRNA transcription compared with high fat high cholesterol group, but also the nuclear level of NF-κB of momordicin treat group were obviously lower than high fat high cholesterol group in artery wall, nuclear translo cation of NF-κB was inhibited by momordicin treating. IκB in high fat high cholesterol group was remarkably reduced compared with normal food group (0.19±0.05 vs 0.74±0.15, P<0.05, n5), but this reduction can be obviously inhibited by momordicin intervention(0.19±0.05 vs 0.36±0.07, P<0.01,n5). Conclusions The role of momordicin anti-inflammatory factors generation is related to inhibiting degradation of IκB, which inhibits nuclear translocation of NF-κB.
2012, 20(4):309-314.
Abstract:AimTo investigate the possible signaling pathway in angiopoietin-1 (Ang-1) regulated inflammatory factors of endothelial progenitor cells.MethodsWe transformated Ang-1 into tumor necrosis factor-α (TNF-α) induced endothelial progenitor cell (EPC), used the specific inhibitor pyrrolidine dithio carbamate (PDTC) to inhibit nuclear factor-kappa B (NF-κB).Western blot was used to detect the protein expression of Ang-1 and NF-κB, real-time polymerase chain reaction (real-time PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression of intercellular cell adhesion molecule-1 (ICAM-1) and vascular cellular adhesion molecule-1 (VCAM-1) in EPC.ResultsAng-1 protein expressed while no significant NF-κB protein expressed in EPC.Real-time PCR and ELISA show that compared to the TNF-α group, the expressions of ICAM-1 and VCAM-1 in Ang-1 group, PDTC group and Ang-1+PDTC group significantly decreased (p<0.05), but there were no significant differences among the three groups (p>0.05).ConclusionsAng-1 maybe affects adhesion molecules expression in TNF-α induced EPC through NF-κB signaling pathway.
2009, 17(5):363-366.
Abstract:Aim To investigate the effect of genistein on activation of nuclear factor kappa B(NF-κB) in vascular smooth muscle cells(VSMC) induced by oxidized low density lipoprotein(ox-LDL) and further examine the activation mechanism. Methods Human umbilical vein smooth muscle cells were pretreated with genistein in different concentrations(10,30,90 μmol/ L) and then treated by 50 mg/ L of ox-LDL for 2 hours.Immunocytochemistry staining was used to detect the nuclear translocation of NF-κB subunit p65.The intracellular level of inhibitory kappa B(IκB),inhibitor of NF-κB,was detected by Western blotting analysis.The levels of nitric oxide(NO) in the culture medium was detected by the method of nitric acid reductase. Results Genistein significantly restrains the nuclear translocation of NF-κB in VSMC induced by ox-LDL;High concentration of genistein increases intracellular IκB-α level and doesnt have significant effect on level of NO in the culture medium. Conclusion Genistein inhibited activation of NF-κB induced by ox-LDL in VSMC.The mechanism is associated with increase of IκB and may not associate with nitric oxide level.
2008, 16(7):527-531.
Abstract:Aim To study the effect of simvastatin on adiponectin(APN)and nuclear factor-kappa B inhibitor kinase(IKK)mRNA expression in adipose tissue of insulin resistant rats.Methods Insulin resistant rat model was induced by high-fat diet feeding.Twenty-five male Wistar rats were randomly divided into two groups fed with either basic chow(n=10)or high-fat diet(n=15)for 10 weeks,then assessed by euglycemic-hyperinsulinemia clamp technique.And rats in high-fat group were randomly divided into simvastatin intervention group(n=5)and non-simvastatin intervention group(n=5).Rats in simvastatin intervention group were fed with simvastatin 10 mg/(kg·d)in gavage.After 5 weeks intervenient treatment,APN and IKK mRNA expressions were tested with RT-PCR.Results The glucose infusion rate(GIR)in high-fat group decreased significantly compared with basic chow group 0.76±0.28 mg/(kg·min)vs 4.26±0.70 mg/(kg·min),P<0.01.There were no obvious difference on APN mRNA expression of adipose tissue between simvastatin intervention group and non-simvastatin intervention group(0.25±0.12 vs 0.29±0.11,P>0.05);but they decreased significantly compared with basic chow group respectively(1.18±0.12,P<0.05).IKK mRNA expression of adipose tissue in simvastatin intervention group decreased significantly compared with non-simvastatin intervention group(0.15±0.03 vs 1.21±0.03,P<0.05),with no obvious difference from basic chow group(0.15±0.03,P>0.05).Conclusion Simvastatin could not increase APN mRNA expression in adipose tissue of insulin resistant rats induced by high-fat diet feeding.But simvastatin may restore IKK mRNA expression in adipose tissue of insulin resistant rats induced by high-fat diet feeding,which maybe benefit from the anti-inflammatory effect beyond the lipid-lowering effect.
2008, 16(12):928-932.
Abstract:Aim To observe the therapeutic effect of soybean isoflavones (SI) on the expression of peroxisome proliferator activated receptor α (PPARα), nuclear factor-kappa B(NF-κB), vascular cell adhesion molecule-1(VCAM-1), and investigate its mechanism of anti-atherosclerosis in metabolic syndrome rats. Methods Sixty male SD rats were randomly divided into normal diet group (n=10)and high-fat diet group (n=50)to induce metabolic syndrome model. Rats in high-fat diet group were fed with high-lipid high-salt high-sugar food for 20 weeks. After 20 weeks 37 metabolic syndrome rats were got successfully, and then these 37 metabolic syndrome rats were randomly divided into model group(n=9), fenofibrate-treated group(n=8), low dose (90 mg/kg) SI-treated group (n=10) and high dose (360 mg/kg) SI-treated group(n=10). Blood lipid level was measured after 4 weeks and mRNA level of PPARα of rat liver was determined using fluorescent quantitative PCR (FQ-PCR). The expressions of NF-κB and VCAM-1 of rat aorta were observed by immunohistochemistry. Results In high dose SI-treatment group, the low density lipoprotein cholesterol (LDLC),triglyceride (TG) and total cholesterol (TC) levels were significantly lower than those in model group(P<0.01), while the expression of PPARα was increased compared with model group. The expressions of NF-κB and VCAM-1 in model group were higher than those in control group. Compared with model group, high-dose SI decreased the expressions of NF-κB and VCAM-1 on the rat aorta. Conclusion SI can regulate the blood lipid level in metabolic syndrome rat, reduce the level of atherosclerosis factor, and can effectively stop the formation of atherosclerosis. Its mechanism may be associated with up-regulating the expression of PPARα mRNA by SI.
2005, 13(4):464-466.
Abstract:Aim To investigate whether hyperhomocysteinemia could induce vascular inflammatory response in aorta of rabbit. Methods Twenty New Zealand white rabbits were ramdomly divided into control group and methionine group. After twelve weeks, the level of homocysteine (Hcy) were measured, aortic lesion were obsersved, Interleukin-8 (IL-8) were determined by enzyme-linked immunosorbent assay (ELISA), the expression and activation of nuclear factor-κB (NF-κB) were studied by immunohistochemistry. Results Hyperhomocysteinemia was induced after high methionine diet, aortic endotheliocyte came off and local intima thickened. The production of IL-8 and activation of NF-κB were increased by hyperhomocysteinemia. Conclusions Hyperhomocysteinemia can induce vascular inflammatory response. Inflammatory response is one of potential cellular mechanisms by which hyperhomocysteinemia accelerates atherosclerosis.
2004, 12(6):707-709.
Abstract:Aim To measure the activity of nuclear factor-κB (NF-κB) of the peripheral blood mononucleus cells (PBMC) of patient which had acute coronary syndrome (ACS) or stable angina or hypertension, study its relationship with C-reactive protein (CRP), plaque stability and determine its value in the diagnosis of ACS. Methods We compared the results of the activity of nuclear factor-κB and levels of CRP in ACS aged 48-80 years with that of the stable angina (SA) or hypertension as controls. Results OD of NF-κB p50 of PBMC and CRP in ACS group on admission was significantly higher than that in SA group and hypertension group (p<0.01), OD of NF-κB p50 of PBMC and CRP in AMI group on admission was significantly higher than that in UA group (p<0.05). OD of NF-κB p50 of PBMC and CRP in SA group had no difference from that in hypertension group. After two weeks, OD of NF-κB p50 of PBMC in ACS group was significantly decreased compared with that on admission (p<0.01). There was no difference between SA group and hypertension group. There was a positive correlation between OD of NF-κB p50 of PBMC and blood glucose level (r=0.512, p<0.01) in ACS group. OD of NF-κB p50 of PBMC also had a positive correlation with serum CRP level(r=0.771, p<0.01). Conclusions The activation of NF-κB in peripheral blood mononucleus cells may be a predictor of the coronary plaque instability and disruption, and it might be helpful in the diagnosis of ACS.
2002, 10(4):312-315.
Abstract:Aim In order to study the relationship bewteen activity of NF-κB and the expressions of monocyte chemotactic protein-1 (MCP-1) and protein kinase C (PKCα) and atherosclerosis (As) plaques of different stages. Methods Using electrophoretic mobility shift assay (EMSA), immunohistochemitry and in situ hybridyzation,We study activity of NF-κB and the expressions of monocyte chemotactic protein-1 (MCP-1) and protein kinase C (PKCα) varied in atherosclerosis (As)plaques of different stages. Results The resultes confirmed that the activity of NF-κB as well as the expressions of monocyte chemotactic protein-1 (MCP-1) and protein kinase C (PKCα) in As plaque was increased in a time dependent manner as compared with those in control group (p<0.05). Which was consistent with the increaceing of activity of NF-κB and up-regulated in the expressions of MCP-1and PKCα. Conclusion All of these indicated that NF-κB activation, the up-regulated expressions of MCP-1and PKCα may take part in pathogenesis and development of As.
2002, 10(4):320-323.
Abstract:Aim To evaluate the effect of lipopolysaccharide on human monocytes tumor necrosis factor α production and nuclear factor kappa B activation. Methods Monocytes were isolated by Ficoll density gradient from blood of healthy volunteers. The amount of tumor necrosis factor α protein in culture medium was quantified by using tumor necrosis factor α ELISA systems. Nuclear factor kappa B translocation in monocytes was observed by immunohistochemistry assay. Results We find that nuclear factor kappa B was activated after being stimulated for 15 minutes and reached peak at 1 hour. Lipopolysaccharide (1 μg/L~1 mg/L) induced monocytes tumor necrosis factor α production in a dose-dependent manner. Tumor necrosis factor α protein in medium could be detected at 1 hours, and achieve peak at 6 hour. Nuclear factor kappa B activation before tumor necrosis factor α production in monocytes. Conclusions Lipopolysaccharide induced pro-inflammation cytokine production at least in part through nuclear factor kappa B activation in monocytes.