Objective To investigate influence of genders on the activity of nuclear factor-kappa B (NF-κB) in lungs of endotoxemic rats. Methods Twenty female and 20 male Wistar rats were randomly divided into four groups as follow: female control group (n=10), male control group (n=10), male endotoxemic group (n=10), and female endotoxemic group (n=10). The endotoxemic rats model was made by injecting lipopolysaccharide (5 mg/kg) into the abdominal cavity. Tissue samples were collected from the lungs in different groups and electrophoresis mobility shift assay was used to measure the activity of NF-κB. The levels of serum TNF-α and estrogen were measured at the same time. Results There was no significant difference between the activities of NF-κB in male and female control groups (1.33±0.24 vs 1.47±0.40）, and there was also no significant difference between other items in these groups as well (Pgt;0.05). Yet, the activity of NF-κB (female: 12.10±2.89; male: 19.53±2.12) and the level of TNF-α 〔female: (4.10±0.72) ng/ml; male: (6.37±1.29) ng/ml〕 were significantly increased after injection of lipopolysaccharide (Plt;0.01), and the indices in female group were significantly lower than those in male group (Plt;0.01). Correlation analysis showed that there was a positive relation between the activity of NF-κB in lungs and the level of TNF-α (female: r=0.921 1, P=0.013; male: r=0.907 2, P=0.017), and there was a negative correlation between the activity of NF-κB and the level of estrogen (female: r=-0.887 5, P=0.017; male: r=0.872 3, P=0.022) in both male endotoxemic group and female endotoxemic group (Plt;0.05). Conclusion Gender may be one of the factors that influence the activity of NF-κB in the lungs of endotoxemic rats. While on the other hand, endogenous estrogen may protect the lungs of endotoxemic rats from injury by inhibiting the activity of NF-κB.
Objective To study the expression of nuclear factor-κBp65 (NF-κBp65) in gastric carcinoma and its relationship with vascular endothelial growth factor (VEGF). Methods The expression of NF-κBp65 and VEGF in 56 gastric carcinomas was detected with immunohistochemistry and compared with benign tissues. Results The positive rates of NF-κBp65 and VEGF in 56 gastric carcinomas were 62.5% and 76.8% respectively,and which were higher than those of gastric mucosal atypical hyperplasia (33.3% and 44.4%) and the normal gastric mucosa(0 and 8.3%) （P＜0.05,P＜0.01）.It was found that there was relationship between the expression of NF-κBp65 and the clinical stage, invasion depth of tumor and lymph node metastasis (P＜0.05),but there was no relation to the historica type (Pgt;0.05). There was positive correlation between NF-κBp65 and VEGF expression （r=0.36,P＜0.01）. Conclusion NF-κBp65 may play an important role in the development of gastric carcinoma by up-regulate the expression of VEGF.
ObjectiveTo discuss the role of nuclear factor-kappa B in restenosis after angioplasty.MethodsRelated literatures of recent 5 years were reviewed.ResultsNuclear factor-kappa B could lead to hyperplasia of vascular intima which resulted from proliferation and decrease of apoptosis of vascular smooth muscle cells.ConclusionNuclear factor-kappa B plays an important role in restenosis after angioplasty.
Objective To investigate the effects of nuclear factor kappa B decoy oligodeoxynucleotides ( NF-κB decoy ODN) transfection on biological characteristics of mature dendritic cells ( mDCs) in mice. Methods Immature DCs were harvested from Balb / c mice bone marrow, followed by the incubation with antigen OVA and LPS, and mature DCs were evaluated by the expressions of CD11c and MHC-Ⅱ detected by FACS. Mature DCs were transfected with NF-κB decoy ODN and the changes of NF-κB activity after the transfection were detected by EMSA. The expressions of the costimulatory molecules( CD40,CD80 and CD86) on DCs were detected by FACS and the proliferation of T cells was tested by mixed lymphocyte reaction( MLR) . Results The mature DCs were cultured successfully. The NF-κB activity of NF-κB decoy ODN transfected DCs was decreased significantly( P lt; 0. 05) . There was no difference in the expressions of CD40 and CD80, but the expression of CD86 was decreased significantly in NF-κB decoy ODN transfection group( P lt; 0. 05) . MLR test showed that the proliferation of T lymphocyte cells was inhibited by NF-κB decoy ODN transfected DCs, but was stimulated bly by the DCs of other groups. Conclusions Mature DCs transfected with NF-κB decoy ODN could inhibit the proliferation and activation of antigenspecical T cells, which was probably related to the down-regulation of CD86 on DCs. This modified DCs might be a promising vaccine for the treatment of asthma in the future.
Abstract: Objective To study the preventive effect of n-3 polyunsaturated fatty acids on allograft arteriosclerosis. Methods Arterial homeotransplant model were created with 480 rats which were divided into four groups. Control group, no n-3 lyunsaturated fatty acids were taken. Group A, n-3 polyunsaturated fatty acids were taken for two weeks before operation with the dose of EPA 600mg/kg. Group B, 300 mg/kg and group C 150 mg/kg were taken respectively. The recipient’s transplanted vessel was excised after 1,7,14,21and 28 days respectively. The tissue pathological variations, ultrastructure variations and expression variations of intercellular adhesion molecule-1(ICAM-1), vascular cell adhesion molecule-1（VCAM-1）, nuclear factorkappa B（NF-κB） had been observed. Results The pathological changes occurred 7 days after operation in control group and were most prominent on the 28th day, blood vessels were obstructed and the expressions of ICAM-1, VCAM1,NF-κB were markedly intensified than those of group A, B, C (Plt;0.05). The pathological variations of transplanted vessel in group A, B, C occurred later than those in control group. The nonobstruction rates in group A, B, C were better than that in control group. The expressions of ICAM-1， VCAM-1， NF-κB in control group were ber than those in group A， B， C (Plt;0.05). The expressions of ICAM-1， VCAM-1， NF-κB after 1 day or 7 days demonstrated no statistically significant change in group A， B， C （Pgt;0.05）. The preventive effect for allograft vessel atheromatosis in group A and group B was ber than that in group C after 14, 21 and 28d (Plt;0.05). There were no significant difference between group A and group B (Pgt; 0.05). Conclusion The n-3 polyunsaturated fatty acids can prevent the allograft vessel atheromatosis, the most effective dose of n-3 polyunsaturated fatty acids is 300 mg/kg.
Objective To observe the effects of exogenous pulmonary surfactant (PS) on ventilation-induced lung injury (VILI) in rats, and to investigate its possible mechanisms. Methods A total of 40 Wistar rats were divided into 4 groups with randomized blocks method: control group, high tidal volume (HV) group, VILI group, and PS group, with 10 rats in each group. The control group was subjected to identical surgical procedure but was never ventilated. After 30 min of mechanical ventilation (MV) with Vt 45 ml/kg, the rats in HV group were killed immediately; rats in the VILI group were continually ventilated for up to 150 min with Vt 16 ml/kg; in the PS group, 100 mg/kg of PS administered intratracheally and with the same settings as VILI group. Mean artery pressure (MAP), blood gas analysis, lung wet to dry weight ratios (W/D), thorax-lung compliance, and cell counts in bronchoalveolar lavage fluid (BALF) were determined. Nuclear factor-κB(NF-κB) activity in lungs was measured by enzyme-linked immunosorbent assay (ELISA), interleukin-8(IL-8) in serum and BALF was determined by radioimmunoassay (RIA). Pathological examination of the lung was performed. Results Injurious ventilation significantly decreased MAP and PaO2/FiO2, but increased NF-κB activity and W/D. MAP and PaO2/FiO2 improved, but NF-κB activity, IL-8 in serum and BALF, and cell counts in BALF reduced significantly in PS group compared with those in VILI group. Histological studies showed reduced pulmonary edema and atelectasis in the PS group. Conclusion PS administered intratracheally can suppress the increased activity of NF-κB induced by VILI, exogenous PS can be used to treat VILI.
Objective To observed the effect of IL-1β on expression of caudal-related homeobox gene 1 (CDX1) mRNA and mucoprotein 2 (MUC2) mRNA in cultured human gastric epithelial cells GES-1, and to investigate the underlying signal transduction pathways. Methods ①GES-1 cell was activated with IL-1β of different concentrations and time, the expression levels of CDX1 mRNA and MUC2 mRNA were detected by using real-time PCR. ②GES-1 cell was pretreated with PDTC, a NF-κB inhibitor, for 1 h prior to the addition of IL-1β, then the expressions of CDX1 mRNA and MUC2 mRNA were measured. Results Both CDX1 mRNA and MUC2 mRNA were not examined in GES-1 cell under normal culture conditions. But they could be induced by IL-1β with a dose-dependent manner in a concentration range (P＜0.05); 8 h after treatment with IL-1β, the peak values of the expression levels of CDX1 mRNA and MUC2 mRNA were reached (P＜0.05), then declined gradually. When pre-incubated with NF-κB inhibitor PDTC, the expression levels of CDX1 mRNA and MUC2 mRNA were significantly decreased (P＜0.05). Conclusion IL-1β significantly induces the expressions of CDX1 mRNA and MUC2 mRNA in cultured human gastric epithelial cell GES-1 through the NF-κB signal pathway, which indicates that IL-1β plays a role in the process of intestinal metaplasia.
Objective To observe the effect of epidermal growth factor (EGF) on the proliferation, adhesion, invasiveness and the activation of nuclear factor-κB (NF-κB), matrix metalloproteinases (MMPs) expression and explore related mechanisms in pancreatic cancer cells. Methods Cell invasion assay, proliferation assay and adhesion assay were used to examine the proliferation, adhesion and invasiveness of pancreatic cancer cells, respectively. NF-κB activity was detected by electrophoretic mobility shift assay (EMSA), and MMPs protein and mRNA expressions were investigated by gelatin zymography, Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results EGF increased the invasiveness of pancreatic cancer cell in a dose-dependent manner (P＜0.05), but did not affect cell proliferation or adhesion. The expressions of MMP-9 mRNA and protein significantly increased after induction by EGF and were highest when EGF concentration was 50 ng/ml, while there was no effect on the expressions of MMP-2 mRNA and protein. Furthermore, NF-κB activity increased with increased concentration of EGF in a concentration-dependent manner (P＜0.05). In addition, NF-κB activity and the expressions of MMP-9 mRNA and protein by pretreatment with both pyrrolidine dithiocarbamate (PDTC) and EGF decreased when compared that by pretreatment with EGF alone. The invasiveness of pancreatic cancer cell by pretreatment with both PDTC and EGF decreased when compared that by pretreatment with EGF alone and nothing (P＜0.05).Conclusion The findings indicate that the NF-κB-mediated MMP-9 induction is essential for EGF-induced invasiveness in pancreatic cancer cells, which can be inhibited by PDTC.
【 Abstract 】 Objective Overexpressions of epidermal growth factor (EGF) and EGF receptor have been associated with progression and invasive phenotype of pancreatic cancer. However, the underlying molecular mechanism by which EGF worked in pancreatic cancer cells has not been completely understood. In this study, effect of EGF on the invasion and metastasis of pancreatic cancer cells and its regulatory mechanism were investigated. Methods The effects of EGF on the proliferation, adhesion and invasion of pancreatic cancer cells were detected by WST-1 proliferation assay, adhesion assay and invasive assay, respectively. The activity and expression of MMP-2 and MMP-9 were examined by zymography, Western blot and RT-PCR, respectively. The activity of NF- κ B was examined by EMSA. Results EGF could significantly promote the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion. The expressions of NF- κ B and MMP-9 were significantly increased by EGF, but EGF did not affect the activity and expression of MMP-2. Furthermore, EGF stimulated the NF- κ B binding activity. Pretreatment with NF- κ B inhibitors, pyrrolidine dithiocarbamate (PDTC), could significantly inhibit the activity of NF- κ B induced by EGF. Meanwhile, the EGF-induced expression and activity of MMP-9, as well as cell invasiveness were also inhibited by NF- κ B inhibitor. Conclusion EGF could increase the expression and promote the invasiveness of MMP-9 via the activation of NF- κ B in pancreatic cancer cells, which implies that NF- κ B inhibitant, such as PDTC, may diminish the invasiveness of pancreatic cancer cells.
Objective To investigate the expression changes of nuclear factor kappa B (NF-κB) and matrix metalloproteinase-9 (MMP-9) in the cultured hepatocellular carcinoma cells 9204 (HCC9204) transfected with inhibitory kappa B alpha（IκB-α)vector. Methods After pcDNA3-IκB-α vector and pcDNA3 were transfected into HCC9204 by lipofectamine method, Western-blot and RT-PCR analysis were used to detect the expressions of NF-κB and MMP-9. Migration and invasion of tumor cells were assayed by fundus membrane invaded by them. Results When pcDNA3-IκB-α was transfected into HCC9204, the expression of NF-κB was decreased at the protein level, and the expression of MMP-9 mRNA and the invision and metastasis ability of transfected cells were obviously decreased. Conclusion When the activity of NF-κB is inhibited, the ability of invasion and metastasis in HCC9204 cells decrease, which could be related to the decreased the expression of MMP-9.