共查询到20条相似文献，搜索用时 15 毫秒
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AIM: To investigate the effect of inhibiting high-mobility group box protein 1 (HMGB1) expression on the viability and apoptosis of hemangioma endothelial cells (HemECs). METHODS: Human HemECs were isolated and cultured, and HMGB1 small interfering RNA (HMGB1-siRNA) was transfected into the cells. The cell viability was detected by CCK-8 assay. The apoptosis and reactive oxygen species (ROS) content were analyzed by flow cytometry. The protein levels of HMGB1, NF-κB p65, p-IκBα, cyclin D1 and survivin were determined by Western blot. RESULTS: The protein expression of HMGB1 in the HemECs transfected with HMGB1-siRNA was significantly lower than that in blank control group (P<0.05). Compared with NC group, the cell viability was decreased significantly in the HemECs transfected with HMGB1-siRNA, the apoptotic rate was significantly increased, the content of ROS increased significantly, and the protein levels of NF-κB p65, p-IκBα, cyclin D1 and survivin were significantly decreased (P<0.05). After exposure to NF-κB signaling pathway inhibitor PDTC, the cell viability was inhibited, the apoptosis was increased, ROS content, and the protein levels of NF-κB p65, p-IκBα, cyclin D1 and survivin were down-regulated significantly, as compared with si-HMGB1 group (P<0.05). CONCLUSION: Inhibition of HMGB1 reduces the viability of HemECs and induces apoptosis by increasing the content of ROS and down-regulating the activity of NF-κB signaling pathway. 相似文献
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AIM: To observe the effects of diterpenoid C from ether extract of Radix Curcumae (RC) on the activity of nuclear factor-κB in human gastric adenocarcinoma SGC7901 cells stimulated with lipopolysaccharide (LPS). METHODS: SGC7901 cells were normally cultured, induced by LPS, or treated with LPS plus RC. The protein expression of IKKα, IKKβ, p65, phosphorylated p65 and phosphorylated IκBα was assayed by Western blotting. NF-κB DNA binding activity was determined by electrophoretic mobility shift assay. RESULTS: RC reduced the protein expression of IKKα, IKKβ, p65, phosphorylated p65 and phosphorylated IκBα induced by LPS. NF-κB DNA binding activity increased much greatly by LPS stimulation, while RC resisted the action of LPS. CONCLUSION: RC may attenuate the secretion of inflammatory cytokines by inhibiting the activation of NF-κB signaling pathway. 相似文献
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AIM:To investigate the effects of 1,25-dihydroxyvitamin D3 ［1,25-(OH)2D3］ on nuclear factor kappa B (NF-κB) signaling pathway in passively sensitized human airway smooth muscle cells (HASMCs). METHODS:HASMCs were passively sensitized with 10% serum from asthmatic patients. 1,25-(OH)2D3 was used as an interfering factor. Electrophoretic mobility shift assay (EMSA) was used to detect the DNA-binding activity of NF-κB. Immunocytochemical staining was used to observe the nuclear translocation of NF-κB p65. Western blotting was used for IκBα and phosphorylated IκBα protein detection. Real-time fluorescence quantitative PCR was used to determine vitamin D receptor (VDR), vitamin D 24-hydroxylase (CYP24) and IκBα mRNA expression. The mRNA expression of IκBα in HASMCs after actinomycin D treatment was also determined. RESULTS:(1) 1,25-(OH)2D3 significantly attenuated the DNA-binding activity of NF-κB and the nuclear translocation of NF-κB p65 in HASMCs passively sensitized by asthmatic serum. (2) 1,25-(OH)2D3 enhanced IκBα mRNA stability and inhibited IκBα protein phosphorylation in passively sensitized HASMCs, thus increasing IκBα expression in these HASMCs. (3) 1,25-(OH)2D3 up-regulated VDR mRNA level and evoked its functional response in passively sensitized HASMCs. CONCLUSION: 1,25-(OH)2D3 enhanced the expression of IκBα and therefore inhibited NF-κB signaling passway in HASMCs. This effect may be dependent on VDR, and responsible for the inhibitory effect of 1,25-(OH)2D3 on passively sensitized HASMCs. 相似文献
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AIM: To observe the effects of TNF-α/nuclear factor-κB(NF-κB)/matrix metalloproteinase-2(MMP-2) pathway on the expression of MMP-2 in the mice with viral myocarditis. METHODS: Six-week-old inbred male mice were randomly assigned to control and myocarditis group. The mice in myocarditis group and control group were intraperitoneally inoculated with 0.1 mL 10-5.69 TCID50/mL coxsackievirus B3 and vehicle (PBS), respectively. Ten mice were sacrificed at the 4th and 10th days after injection. The blood and heart specimens were harvested. The serum content of TNF-α was measured by ELISA. The myocardial levels of MMP-2, NF-κB p65 and IκBα were determined by Western blot. RESULTS: Compared with control group, the protein expression of MMP-2 and NF-κB p65 in the myocardium and the serum content of TNF-α were significantly increased in myocarditis group (P<0.05). The protein expression of IκBα was lower in myocarditis group than that in control group (P<0.05).CONCLUSION: TNF-α, NF-κB p65 and MMP-2 were higher in the mice with acute viral myocarditis. The increased expression of them might be involved in the pathogenesis of viral myocarditis. 相似文献
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IκB is an inhibitor of nuclear factor-kappa B (NF-κB) and presents in the majority of cells. Eight structurally related members of the mammalian IκB family have been described:IκBα, IκBβ, IκBε, Bcl-3, IκBγ, IκBδ, p100 and p105. The ankyrin repeat domain of IκB can interact with NF-κB, and sequester NF-κB in the cytoplasm as inactive complexes. Most recently, IκBα has been found to inhibit p53 tumor suppressor protein by binding p53 to form a cytoplasmic p53·IκBα complex and studies have shown that p100 profoundly sensitizes cells to death-receptor-mediated apoptosis. The current review is to describe the members of IκB family, their related signaling pathways, and their application in tumor therapy. 相似文献
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YANG Zhou GANG Hai-ju QIN Xian-peng JIA Gui-qing WANG Bo YANG Chun ZHAO Gao-ping 《园艺学报》2018,34(12):2153-2159
AIM:To investigate the effect of Krüppel-like factor 4 (KLF4) on the viability, apoptosis and cisplatin chemosensitivity of colorectal cancer cells. METHODS:KLF4 expression in colorectal cancer cell lines Caco2, SW480 and HCT116 was detected by Western blot. The SW480 cells were divided into pcDNA3.1 group (transfected with pcDNA3.1 empty plasmid), pcDNA3.1-KLF4 group (transfected with pcDNA3.1-KLF4 expression plasmid) and pcDNA3.1-KLF4+cisplatin group (treated with 1 mg/L cisplatin for 48 h after pcDNA3.1-KLF4 was transfected into SW480 cells). The protein levels of KLF4, p-IκBα, cyclin D1 and survivin were determined by Western blot. The cell viability was measured by CCK-8 assay. The apoptotic rate was analyzed by flow cytometry. The content of reactive oxygen species(ROS) was measured by DCFH-DA probe. RESULTS:The expression of KLF4 in the colorectal cancer cells were significantly lower than that in the human colon mucosal epithelial NCM460 cells (P<0.05). Compared with pcDNA3.1 group, the protein expression of KLF4 in pcDNA3.1-KLF4 group was significantly increased (P<0.05). Compared with pcDNA3.1 group, the cell viability and the protein expression of cyclin D1 and survivin were significantly decreased, and the apoptotic rate, the content of ROS and the protein level of p-IκBα were significantly increased in pcDNA3.1 group (P<0.05). Compared with pcDNA3.1-KLF4 group, the cell viability and the expression of cyclin D1 and survivin proteins were significantly decreased, and the apoptotic rate, the content of ROS and the protein level of p-IκBα were significantly increased in pcDNA3.1-KLF4+cisplatin group (P<0.05). CONCLUSION:Upregulation of KLF4 gene expression in colorectal cancer cells reduces the cell viability, induces apoptosis and increases the chemosensitivity of the cells to cisplatin. The mechanism may be related to the enhancement of intracellular ROS content and down-regulaton of the phosphorylation level of IκBα, the key molecule of NF-κB signaling pathway. 相似文献
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HE Fang PENG Jing DENG Xiao-Lu YANG Li-fen WU Li-wen ZHANG Ci-liu YIN Fei 《园艺学报》2011,27(6):1041-1047
AIM: To investigate the role of nuclear factor kappa B (NF-κB) signaling in the changes of permeability in brain-derived microvascular endothelial (bEnd.3) cells induced by lipopolysaccharide (LPS).METHODS: The bEnd.3 cells were randomly divided into 3 groups: bEnd.3 group, bEnd.3/vector group and bEnd.3/muIκBα group. The cells in the latter 2 groups were transfected with pcDNA3.1hygro and DNMu-IκBα (a dominant-negative mutant of IκB) plasmids, respectively. All the cells were exposed to LPS. The activity of NF-κB, monolayer barrier integrity and F-actin distribution were detected by luciferase reporter assay, transendothelial electrical resistance (TEER) assay and rhodamine-phalloidin staining, respectively. The expression of tight junction proteins (ZO-1 and claudin-5) and phosphorylation of myosin light chain (MLC) were determined using Western blotting.RESULTS: In bEnd.3 group and bEnd.3/vector group, the NF-κB activity began to increase obviously as early as 0.5 h after pretreatment with LPS. LPS decreased TEER, and induced F-actin rearrangement and ZO-1 down-regulation in 3 h. Incubation of the cells with LPS for 12 h induced the most significant disruptive effects on the permeability and tight junctions. Moreover, high expression of phosphorylated MLC accompanied with the early damages of tight junctions was observed. However, these destabilizing alterations were suppressed in bEnd.3/muIκBα group by the inhibition of NF-κB activity.CONCLUSION: LPS induces hyperpermeability in brain microvascular endothelial cells. The functions of NF-κB signaling are related to influencing disruptions of tight junctions by regulating the phosphorylation of MLC. 相似文献
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AIM: To investigate the effects of burn sera on IκBα degradation, NF-κB activation in peripheral blood monocytes (PBMCs) in order to explore the role of burn sera on activation of monocytes. METHODS: PBMCs isolated from healthy volunteers were stimulated by sera from healthy volunteers and burn patients and by burn sera together with PDTC (pyrrolidine dithioncarbamate). Activation of monocytic NF-κB was tested by electrophoretic mobility shift assay (EMSA) and the degradation of monocytic IκBα was determined by Western blotting. RESULTS: When compared to that in control group, cytosolic IκBα degradation occurred within 30 min after PBMCs stimulated by burn sera, and peaked at 60 min. But IκBα gradually recovered in the cytoplasm after 2 h of stimulation. Meanwhile, activity of monocytic NF-κB was markedly increased, reached the peak at 30 min to 60 min after stimulation, and gradually decreased after 2 h of stimulation. PDTC (an antioxidants) effectively inhibited the monocytic IκBα degradation and activation of NF-κB induced by burn sera. CONCLUSION: Burn sera might induce the degradation of IκBα, then activate NF-κB, which ultimately lead to the secretion of cytokines from the monocytes. 相似文献
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AIM: To investigate the combined effect of octreotide and Dachaihu decoction on the treatment of severe acute pancreatitis(SAP). METHODS: Wistar rats(n=50) were randomly divided into sham group, SAP group, octreotide group, Dachaihu decoction group and combination group. The quantity of ascites was measured. The levels of amylase, alanine aminotransferase and creatinine in the serum were examined. The morphological changes of the pancreatic tissues were observed by HE staining. The activation of NF-κB and IκBα expression were determined by Western blot. The mRNA expression with ICAM-1 and IL-1 was detected by qPCR.RESULTS: Combined treatment with octreotide and Dachaihu decoction effectively reduced the quantity of ascites and the levels of amylase, alanine aminotransferase and creatinine in the serum in SAP rats. Moreover, combined treatment significantly inhibited SAP-induced activation of NF-κB and decrease in IκBα protein expression, accompanied by a decrease in ICAM-1 and IL-1 mRNA expression. CONCLUSION: Combination of octreotide with Dachaihu decoction effectively attenuates SAP by inhibiting NF-κB signaling pathway and ICAM-1 and IL-1 expression. 相似文献
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AIM: To observe the anti-atherosclerosis effect of paeonal (Pae) on the activation of NF-κB and the expression of cell adhesion molecules in human umbilical vein endothelial cells (HUVECs) induced by hyperlipidemic serum. METHODS: Cultured HUVECs were used as target cells. Hyperlipidemic serum was added to the culture medium to establish the injury mode of HUVECs. Methyl thiazolyl tetrazolium (MTT) method was used to examine the cell viability. The mRNA expression of NF-κB p65 was determined by RT-PCR. The protein levels of IκB-α, intercellular cell adhesion molecule-1 (ICAM-1) and E-selectin were detected by Western blotting. RESULTS: After treated with Pae, the cell viability was increased and the morphological changes of HUVECs injured by hyperlipidemic serum trended to normal. The expression of IκB-α in HUVECs injured by hyperlipidemic serum increased, while the expression of NF-κB p65 mRNA, ICAM-1 and E-selectin protein was decreased. CONCLUSION: The anti-atherosclerosis mechanism of paeonal may be related to the inhibitory effect of the natural compound on the pathway of NF-κB/IκB, thereby reducing the expression of ICAM-1 and E-selectin and attenuating the inflammatory reaction in vascellum. 相似文献
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AIM: To investigate the effects of recombinant human interleukin-17A (rhIL-17A) on the viability and apoptosis of human skin keratinocytes and fibroblasts, and to observe the secretion of profibrotic cytokines by fibroblasts. METHODS: Human skin keratinocytes and fibroblasts were treated with different concentrations of rhIL-17A. CCK-8 method was used to test the cell proliferation. The protein expression of nuclear factor-κB/p65 (NF-κB/p65) and IκBα was determined by Western blotting. The cell apoptosis was observed by flow cytometry. The secretion of interleukin-6 and transforming growth factor-β1 in the culture supernatants of fibroblasts was assayed by ELISA. RESULTS: No difference of the keratinocyte numbers between rhIL-17A treatment groups and control group was observed, while the numbers of fibroblasts were higher in rhIL-17A treatment groups than that in control group (P<0.05). The protein expression of NF-κB/p65 increased in fibroblasts with rhIL-17A treatment, while the expression of IκBα decreased. rhIL-17A had no effect on the apoptosis of both keratinocytes and fibroblasts. The secretion of interleukin-6 and transforming growth factor-β1 in fibroblasts increased after treated with rhIL-17A. CONCLUSION: rhIL-17A had no effect on the proliferation of keratinocytes. However, it can enhance the proliferation of fibroblasts. This effect may be attributed to the activation of NF-κB in fibroblasts by interleukin-17. It is possible that rhIL-17A causes the cell proliferation and collagen synthesis by stimulating fibroblasts to secrete interleukin-6 and transforming growth factor-β1. 相似文献
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AIM: To investigate the effect of linarin (LIN) on the migration and invasion abilities of human breast cancer MDA-MB-231 cells and its underlying mechanism. METHODS: MCF-7, MDA-MB-231 and MCF-10A cells were cultured in vitro and treated with LIN at 5, 10, 20, 40, 80 and 160 μmol/L for 24 h, and the cell proliferation was measured by CCK-8 assay and colony formation assay. The protein levels of Snail, E-cadherin, matrix metalloproteinase-9 (MMP-9), IκBα, p-IKKα/β and p-p65 were determined by Western blot. RESULTS: LIN remarkably reduced the viability of MDA-MB-231 cells in a dose-dependent manner (P<0.05), and the IC50 was 55.89 μmol/L for 24 h. LIN decreased the colony formation rate of MDA-MB-231 cells at the concentration of 20 μmol/L (P<0.05). After exposed to LIN at 5 μmol/L and 10 μmol/L for 24 h, the migration and invasion abilities of the MDA-MB-231 cells were significantly reduced (P<0.05), the protein expression levels of E-cadherin and IκBα were up-regulated (P<0.05), the protein expression levels of Snail and MMP-9 were down-regulated (P<0.05), and the phosphorylation levels of IKKα/β and p65 were decreased (P<0.05) in comparison with the control group. Meanwhile, IKK-16 (IKKα/β inhibitor) and PDTC (NF-κB inhibitor) also down-regulated the protein expression levels of Snail and MMP-9 (P<0.05), and up-regulated the protein expression level of E-cadherin (P<0.05). CONCLUSION: LIN down-regulates the protein expression levels of Snail and MMP-9, and up-regulates the protein expression level of E-cadherin most likely through inhibiting IKK/NF-κB signaling pathway, and ultimately lead to decreases in the migration and invasion abilities of MDA-MB-231 cells. 相似文献
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XU Han-shi YANG Xiu-yan LIANG Liu-qin LI Zhi-jiang YANG Xiao YE Yu-jin LI You-ji 《园艺学报》2003,19(10):1305-1310
AIM:To examine whether Akt signal pathway proteins, including Akt, NF-κB and IκBα, are activated in kidney tissue of murine chronic graft-versus-host disease (GvHD) lupus nephritis in vivo, and whether prednisone suppresses activation of them.METHODS:Akt activity and phosphorylated IκBα were detected by Western-blot. Activation of NF-κB was detected by electropheretic mobility shift assay (EMSA). RESULTS:Activity of Akt, NF-κB and phosphorylated IκBα were significantly increased in kidney tissue of murine chronic graft-versus-host disease (GvHD) in 8th week and 12th week after monocell injection, respectively. However, they were no significant elevation in 16th week, when compared with controls. Prednisone treatment significantly prevented the increase in serum anti-dsDNA antibody level, urinary protein excretion and glomerular cell proliferation in GvHD mice, indicating the beneficial effects of prednisone on this model. Prednisone also significantly suppressed the increase in the activities of glomerular Akt, NF-κB and phosphorylated IκBα. CONCLUSION:This study provides the first evidence of marked increase in glomerular Akt-NF-κB signal pathway activities in murine chronic graft-versus-host disease lupus nephritis. The beneficial effect of prednisone on this lupus nephritis model may be partially mediated by the suppression of abnormal Akt- NF-κB activation. 相似文献
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AIM: To investigate the effect of NOD8 on lipopolysaccharide (LPS)-induced releases of nitric oxide (NO), tumor necrosis factor α (TNF-α) and interleukin-1β (IL-1β) in RAW264.7 cells. METHODS: The plasmids of pEGFP-C2 and pEGFP-NOD8 were transfected into RAW264.7 cells respectively. The transfected and non-transfected cells were stimulated by LPS for 0, 6, 12 and 24 h. NO production was evaluated by Griess reagent assay, and the levels of IL-1β and TNF-α were measured by ELISA. The protein expression of NOD8 and the nuclear translocation of nuclear factor κB (NF-κB) p65 subunit were detected by Western blotting. The level of activated caspase-1 was determined by fluorimetric method. RESULTS: Compared with pEGFP-C2 group, the protein expression of NOD8 was significantly elevated in pEGFP-NOD8+LPS group. The releases of NO, IL-1β and TNF-α were obviously increased after RAW264.7 cells were treated with LPS for 6 h, 12 h and 24 h, and while the secretion of NO was significantly reduced in the cells transfected with pEGFP-NOD8 and induced by LPS for 12 h and 24 h, and the release of IL-1β was also significantly reduced at 6 h, 12 h and 24 h. However, no significant difference of TNF-α release was observed between pEGFP-C2+LPS group and pEGFP-NOD8+LPS group. The activation of caspase-1 in RAW264.7 cells stimulated with LPS for 6 h, 12 h and 24 h was markedly increased, and the expression of NF-κB p65 subunit in the cytoplasm was significantly decreased, indicating that p65 nuclear translocation was increased. In addition, the activation of caspase-1 and the nuclear translocation of p65 were significantly inhibited in pEGFP-NOD8+LPS group. CONCLUSION: NOD8 suppresses the releases of LPS-induced NO and IL-1β in RAW264.7 cells by inhibiting the activation of caspase-1 and NF-κB. 相似文献
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LI Yan MA Ming-ming ZHANG Xiao-qiang PAN Wen-fei LIU Xiang-yong WANG Xiao-zhi 《园艺学报》2013,29(11):2088-2091
AIM:To observe the effects of angiopoietin 4 (Ang-4) on lipopolysaccharide (LPS)-induced injury of human umbilical vein endothelial cells (HUVECs). METHODS:The EnVision immunohistochemical method was used to identify the HUVECs. After pre-treated with different doses of Ang-4 for 0.5 h, HUVECs was exposed to LPS at concentration of 10 mg/L for 24 h. The cell viability was evaluated by MTT assay. The content of tumor necrosis factor-alpha (TNF-α) in the supernatant and the concentrations of intracellular and supernatant von Willebrand factor (vWF) were detected by ELISA. The mRNA levels of Toll-like receptor 4 (TLR4), NF-κB p65 and TNF-α were determined by real-time PCR. RESULTS:Factor Ⅷ in the cytoplasm was positive in the HUVECs.Compared with normal group, LPS reduced the cell viability (P<0.01), and significantly increased the secretion of TNF-α and vWF (P<0.01). The mRNA expression of TLR4, NF-κB p65 and TNF-α also increased (P<0.01). Ang-4 at concentration of 100 μg/L enhanced the cell viability (P<0.01), reduced the content of vWF and TNF-α, and inhibited the LPS-induced increases in the mRNA levels of TLR4, NF-κB p65 and TNF-α (P<0.01). CONCLUSION: Ang-4 antagonizes LPS-induced damage in HUVECs by inhibiting TLR4-NF-κB p65-TNF-α signaling pathways. 相似文献