In vitro anticancer property of a novel thalidomide analogue through inhibition of NF-κB activation in HL-60 cells

Aim： To investigate the anticancer property and possible mechanism of action of a novel sugar-substituted thalidomide derivative （STA-35） on HL-60 cells in vitro.
Methods： TNF-α-induced NF-κB activation was determined using a reporter gene assay. The MTT assay was used to measure cytotoxicity of the compound. The appearance of apoptotic Sub-G1 cells was detected by flow cytometry analysis. PARP cleavage and protein expression of NF-κB p65 and its inhibitor IκB were viewed by Western blotting.
Results： STA-35 （1-20 μmol/L） suppressed TNF-α-induced NF-κB activation in transfected cells （HEK293/pNiFty-SEAP） in a dose- （1-20 μmol/L） and time-dependent （0-48 h） manner. It was also shown that STA-35 exerted a dose-dependent inhibitory effect on HL-60 cell proliferation with an IC50 value of 9.05 lamol/L. In addition, STA-35 induced apoptosis in HL-60 cells, as indicated by the appearance of a Sub-G1 peak in the cell cycle distribution, as well as poly ADP-ribose polymerase （PARP） cleavage. Subsequently, both NF-κB p65 and its inhibitor IκB gradually accumulated in cytoplasmic extracts in a dose- and time-dependent manner, indicating the blockage of NF-κB translocation induced by TNF-α from the cytoplasm to the nucleus.
Conclusion： A novel sugar-substituted thalidomide derivative, STA-35, is potent toward HL-60 cells in vitro and induces apoptosis by the suppression of NF-κB activation.     

Effect of α-pinene on nuclear translocation of NF-κB in THP-1 cells

AIM: To study the effects of α-pinene on nuclear translocation of nuclear factor-κB (NF-κB) and the expression of the inhibitor of NF-κB (IκBα) in human monocyte THP-1 cell line. METHODS: THP-1 cells were incubated with α-pinene (1, 10, and 100 mg/L, for 30 min) before being stimulated with lipopolysaccharide (LPS, 1 mg/L, 30 min).The location of NF-κB p65 subunit (NF-κB/p65) in THP-1 cells was detected by immunofluorescence and laser scanning confocal microscope (LSCM). The expression of NF-κB/p65 in nuclei and that of IκBα in cytoplasm were measured by Western-blot analysis. RESULTS: The majority of FITC-labelled NF-κB/p65 was located in the nuclei being stimulated with LPS. Whereas, no such fluorescence was seen in the nuclei of the groups pretreated with α-pinene or control cells, α-Pinene pretreatment decreased the NF-κB/p65 nuclear translocation in LPS-stimulated THP-1 cells, and this effect was dose-dependent, but there was no reaction in LPS-unstimulated THP-1 cells, α-Pinene pretreatment increased IκBα protein level in cytoplasm, compared with that in LPS-stimulated THP-1 cells. CONCLUSION: In a dose-related fashion, α-pinene inhibits the nuclear translocation of NF-κB induced by LPS in THP- 1 cells, and this effect is partly due to the upregulation of IκBα expression.     

Effect and mechanism of baicalein on 2,4,6-trinitrobenzene sulfonic acid-induced experimental colitis of mice北大核心CSCD

OBJECTIVE: To explore the effect and mechanisms of baicalein on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis in mice. METHODS: BALB/c mice were randomly placed into three groups (n=10): normal control group, TNBS group, and TNBS+baicalein (20 mg.kg-1, once per day) group. Mouse colitis was induced by intrarectal injection of TNBS. Baicalein was administered by oral gavage two days prior to TNBS treatment and until the end of the study (a total of 9 d). The colon length was measured before HE staining was performed for histological damage assessment. The remaining colon pieces were collected to measure the content of tumor necrosis factor-α(TNF-α). Lipopolysaccharide (LPS)-stimulated RAW264.7 mouse macrophage was used as a cell model to determine the content of nitric oxide (NO) in cell culture medium, the mRNA levels of TNF-α, interleukin-6(IL-6), IL-1β, inducible nitric oxide synthase(iNOS), cyclooxygenase 2(COX-2) and monocyte chemoattractant protein-1 (MCP-1), and the protein expression of phosphatidylinositol 3-kinase/protein kinase B/nuclear factor-κB (PI3K/AKT/NF-κB) pathway. RESULTS: Baicalein significantly attenuated TNBS-induced colon shortening and histological injury (P<0.05), which was correlated with the decline in the content of TNF-α in the colon. According to the in vivo results, baicalein exposure down-regulated the secretion of NO and the mRNA expression of pro-inflammatory mediators (iNOS, COX-2, MCP-1, TNF-α, IL-1β and IL-6) in LPS-stimulated RAW264.7 cells (P<0.05, P<0.01). Additionally, the phosphorylation/activation of LPS-stimulated PI3K/AKT/NF-κB pathway was inhibited by baicalein treatment. CONCLUSION: The beneficial effect of baicalein in TNBS-induced experimental colitis may be due to PI3K/AKT/NF-κB signaling inhibition.     

Effect ofα-pinene on nuclear translocation of NF-κB in THP-1 cells

AIM: To study the effects of α-pinene on nuclear translocation of nuclear factor-κB (NF-κB) and the expression of the inhibitor of NF-κB (IκBα) in human monocyte THP-1 cell line. METHODS: THP-1 cells were incubated with α-pinene (1, 10, and 100 mg/L, for 30 min) before being stimulated with lipopolysaccharide (LPS, 1 mg/L, 30 min). The location of NF-κB p65 subunit (NF-κB/p65) in THP-1 cells was detected by immunofluorescence and laser scanning confocal microscope (…     

Insulin-like growth factor-1 promotes cell cycle progression via upregulation of cyclin D1 expression through the phosphatidylinositol 3-kinase/nuclear factor-κB signaling pathway in FRTL thyroid cells

Aim： Insulin-like growth factor-1 （IGF-1） is an important hypertrophic and cell cycle progression factor for a number of cell types. It has been proven that IGF-1 is involved in the regulation of thyroid proliferation and cell cycle progression; however, the exact mechanism of this regulation has not been fully elucidated. In the present study, we investigated the effect of IGF-1 on the expression of cyclin D1, an important cell cycle regulatory protein, and a signaling pathway involved in IGF-1＇s effect on cyclinD1 expression in FRTL thyroid cells.
Methods： FRTL thyroid cells were treated with IGF-1 or vector control for 24 h. As appropriate to individual experiments, a phosphatidylinositol 3-kinase （PI3K） inhibitor, LY294002, and/or a nuclear factor-κB （NF-κB） inhibitor, BAY11-7082, were added 1 h prior to IGF-1 treatment. Western blotting was used to detect cyclin D1 protein expression. Immunofluorescence was performed to analyze the expression of IκBα, an NF-κB inhibitory protein. Cell cycle analysis was performed by fluorescence activated cell sorting （FACS）.
Results： IGF-1 increased the cyclin D1 expression in thyroid cells. This increase was blocked by pretreatment with LY294002 or BAY11-7082. Further studies showed that IGF-1 specifically induced NF-κB activity. Treatment with IGF-1 could accelerate cell cycle progression from G0/G1 to S phase, whereas this progression was inhibited by the presence of LY294002 or BAY11-7082.
Conclusion： In summary, the results of the present study show that in FRTL cells, IGF-1 promotes cell cycle progression via an upregulation of cyclin D1 expression, at least partially through the PI3K/NF-κB signaling pathway.     

在患有急性肺损伤的小猪的肺泡巨噬细胞中一氧化氮，表面活性剂和糖皮质激素对核因子-κB和激活蛋白-1的活性的调控作用

AIM： To investigate whether acute lung injury (ALI) in ventilated piglets with bacterial infection affects NF-κB and AP-1 expression in alveolar macrophages (AM) and whether nitric oxide (NO), surfactant (Surf), glucocorticoids (GC) affect NF-κB and AP-1 activation in AM in vivo and in vitro. METHODS: The animals were intraperitoneally injected Escherichia coli, which caused ALI. Nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for the nuclear factor-kappa B (NF-κB) and activation protein-1 (AP-1) expression. Detection of IκB-α protein was from cytoplasmic extract by Western blotting. Immunocytochemistry staining was used for intracellular location of p65 subunits of NF-κB. RESULTS: In ex vivo experiments, strikingly higher expression of NF-κB and AP-1 by EMSA was found 6h after bacterial injection in contrast to the Normal group. In the NO, SNO,and GC groups, markedly attenuated NF-κB and AP-1 activation was observed. The NF-κB and AP-1 activation in Surf group showed lower levels of the expression. Immunoblotting of AM cytoplasmic extract showed low expression of IκB-α protein in the Control and Surf groups. The stronger expression was observed in the NO, GC,and SNO groups. AM of the Control and Surf groups showed intense nuclear staining, with decreased nuclear staining in the NO, GC and SNO groups. In in vitro experiment, it caused a significant increase in NF-κB and AP1 activity in AM 1h after exposure to lipopolysaccharides (LPS). In AM treated by LPS SNP and LPS GC, all showed decrease of DNA binding activity of NF-κB and AP-1 compared to those exposed to LPS Surf. Immunoblotting of AM cytoplasmic extract showed that LPS stimulation of AM resulted in the low expression of IκB-α protein, which was not observed in the presence of SNP and methylprednisolone. However, the surfact antdid not show such effect. LPS Surf-exposed AM had intense nuclear staining, whereas decreased nuclear staining in the LPS NO and LPS GC-treated cultures was found, confirming a decrease in NF-rd3 activity. CONCLUSION:Activation of NF-κB was found in AM of ventilated piglets with bacterial ALI. NO and GS could prevent NF-κB and AP-1 activation in vivo and in vitro. Surfactant has limited effects on NF-κB and AP-1 activity.     

Baicalin inhibits streptozotocin-induced neuroinflammation in Alzheimer' s disease rat model by TLR4/MyD88/NF-κB pathway北大核心CSCD

Aim To investigate the effects of baicalin on the inflammatory response and Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD 88)/nuclear factor kappa B (N F-K B) signaling pathway in Alzheimer' s disease (AD) rat model induced by lateral ventricular injection of streptozotocin (STZ). Methods The AD animal model was constructed by lateral ventricular injection of STZ in SD rats, and divided into sham operation group, model group, low-dose (60 mg -1 • kg-1 • d-1 ) baicalin group, high-dose (120 mg-1 • kg-1 • d-1 ) baicalin group and minocycline group (36 mg-1 • kg-1 • d-1 ). The learning and memory ability of rats were assessed by Morris water maze. The mRNA expressions of tumor necrosis Factor-α(TNF-α), interleukin-6 (IL-6), interleukin-1β(IL-1β) were detected by real-time quantitative PCR, the protein expressions of TLR4, MyD88, N F-Κ B p65, p-NF-κB p65 and nucleus/cytoplasm NF-ΚB p65 were detected by Western blot, and NF-ΚB p65 transfering from cytoplasm to nucleus and the activation of microglia were measured by immunofluorescence. Results Compared with the sham operation group, animals in the model group showed decreased learning and memory capacity, increased mRNA expression of TNF-α, IL-6 and IL-1κ, up-regulated protein expressions of TLR4 and MyD88, the ratio of p-NF-κB p65/NF-κB p65 and the nucleus/cytoplasm ratio of NF-ΚB p65, enhanced expression of NF-KB p65 in nucleus and the activation of microglias. Compared with the model group, baicalin group and minocycline group improved the learning and memory capacity of AD rats, decreased the mRNA expression of TNF-α, IL-6 and IL-1β, down-regulated the protein expressions of TLR4 and MyD88, the p-NF-ΚB P65/NF-KB p65 ratio and the nucleus/cytoplasm ratio of NF-KB p65, and reduced the expression of NF-KB p65 in nucleus and the activation of microglias. Conclusion Baicalin may exert neuroprotective effects by inhibiting the TLR4/MyD88/NF-KB signaling pathway and neuroinflammation response in the AD brain. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Effect of phillygenin on inflammatory response of A549 cells induced by lipopolysaccharide and normal human plasma北大核心CSCD

Aim To investigate the effect of phillygenin ( PHI) on lipopolysacchride ( LPS) and normal human plasma ( NHP) induced inflammatory injury on alveolar type II epithelial A549 cells and the related mechanism. Methods A549 cells were exposured to 1 mg • L -1 of LPS and 5% NHP to build the inflammatory injury model. The effects of PHI on cell viability, cell number, area, morphology, and DNA content of A549 cells were detected by thiazole blue colorimetric (MTT) and Hoechst 33342 staining. The A549 cells were pretreated with PHI for 2 h, and then exposed to 1 mg • L -1 of LPS and 5% NHP. The contents of inflammatory cytokines and NF-κB p65 were determined by ELISA and quantitative real-time PCR. The transfer of NF-κB p65 from cytoplasm to nucleus was detected by cellular immunofluorescence. The protein expres¬sions of NF-κB and MAPK signaling pathways were de¬tected by Western blot. Results The contents of IL-6 and IL-8 and the transfer of NF-κB p65 from cytoplasm to nucleus were significantly up-regulated in LPS combined with NHP co-stimulated group. PHI at 1, 10, 50, and 100 jjimol • L -1decreased the mRNA and protein expressions of IL-6 and IL-8 , inhibited the transfer of NF-κB p65 from cytoplasm to nucleus, reduced the NF-κB p65 mRNA level, down-regulated the phospho-rylation levels of IκBα, NF-κB p65, p38, JNK, and ERK, and up-regulated the protein expression of IκBα in a dose-dependent manner. Conclusions LPS combined with NHP successfully induces inflammatory injury in alveolar type II epithelial A549 cells. PHI could improve the inflammatory response by inhibiting the activation of LPS-TLR4-NF-κB/MAPK signaling pathways. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

Salvianic acid A inhibits induction of inflammatory mediators by blocking Nuclear Factor-κB activation in macrophages

Objective To investigate the anti-inflammation effect and possible mechanism of Salvianic acid A(SAA)in mouse peritoneal macrophages.Methods Peritoneal macrophages were obtained from BALB/c mice.LPS induced nitric oxide(NO),tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)in supernatant,protein expression of inducible nitric oxide synthase(iNOS),matrix metalloproteinase-9(MMP-9)and activation of nuclear factor-kappa B(NF-κB)in the extract were measured.Results SAA strongly inhibited the excessive production of NO,TNF-α and IL-6 in LPS-induced peritoneal macrophages in a concentration-dependent manner and blocked the expression of iNOS and MMP-9.Treatment with LPS alone increased the translocation of NF-κB(p65)from cytosol to the nucleus,but the SAA inhibited the translocation of NF-κB(p65).Conclusions The results showed that SAA had strong anti-inflammatory effects in LPS-stimulated peritoneal macrophages.The important mechanism is due to its inhibition of NF-κB activation.     

Regulation of activity of nuclear factor-kB and activator protein-1 by nitric oxide, surfactant and glucocorticoids in alveolar macrophages from piglets with acute lung injury

AIM: To investigate whether acute lung injury (ALl) in ventilated piglets with bacterial infection affects NF-κB and AP-1 expression in alveolar macrophages (AM) and whether nitric oxide (NO), surfactant (Surf), glucocorticoids (GC) affect NF-κB and AP-1 activation in AM in vivo and in vitro. METHODS: The animals were intraperitoneally injected Escherichia coli, which caused ALl. Nuclear extracts of AM were analyzed by electrophoretic mobility shift assay (EMSA) for the nuclear factor-kappa B (NF-κB) and activation protein-1 (AP-1) expression. Detection of IκB-α protein was from cytoplasmic extract by Western blotting. Immunocytochemistry staining was used for intracellular location of p65 subunits of NF-κB. RESULTS: In ex vivo experiments, strikingly higher expression of NF-κB and AP-1 by EMSA was found 6 h after bacterial injection in contrast to the Normal group. In the NO, SNO, and GC groups, markedly attenuated NF-κB and AP-1 activation was observed. The NF-κB and AP-1 activation in Surf g     

染料木黄酮对哮喘患者核因子-κB表达和肿瘤坏死因子-α分泌的影响

目的:探讨染料木黄酮对支气管哮喘患者外周血单个核细胞(PBMCs)核因子-κB(NF-κB)表达及肿瘤坏死因子-α(TNF-α)分泌的作用.方法:32例支气管哮喘急性发作期患者及31例健康对照者的PBMCs,均分空白对照、地塞米松和染料木黄酮3组进行研究.NF-κB表达由免疫组化染色检测,TNF-α含量由放射免疫法测定.结果:哮喘患者PBMCsNF-κB胞核染色阳性率及培养上清TNF-α含量明显高于健康对照者(P＜0.01),且二者呈显著正相关(P＜0.01).染料木黄酮抑制哮喘组NF-κB高表达及TNF-α高分泌,与哮喘空白对照组比较二者均有统计学意义(P＜0.01),但其抑制作用弱于地塞米松,且哮喘染料木黄酮组PBMCs NF-κB胞核染色阳性率与TNF-α含量呈显著正相关(P＜0.01).结论:哮喘患者NF-κB表达增高,TNF-α分泌增多.染料木黄酮可抑制哮喘患者PBMCs NF-κB的高表达及TNF-α的高分泌,对哮喘可能有潜在的治疗作用.     

FK228对HepG2细胞NF-κB活化及炎症因子转录的影响

目的:研究FK228对TNF-α诱导的人肝癌细胞HepG2核转录因子κB(nuclear factor-κB,NF-κB)活化及炎症因子IL-6、IL-8转录的影响。方法:培养的HepG2细胞分为对照组、TNF-α刺激组和FK228干预组。分别用TNF-α刺激和FK228+TNF-α共同作用,免疫印迹(Western blot)法分析细胞核中NF-κBp65及其细胞浆中抑制因子IκBα的表达;RT-PCR对炎症因子IL-6、IL-8 mRNA作半定量分析。结果:FK228(4～32μg·L~(-1))干预组与TNF-α刺激组比较,细胞核内NF-κB显著减少(P<0.01);FK228(8～32μg·L~(-1))减少胞浆中IκBα的降解且各组之间差异有统计学意义(P<0.01);FK228降低TNF-α诱导的IL-6、IL-8 mRNA表达,FK228干预组与TNF-α刺激组相比,差异具统计学意义(P<0.01)。结论:FK228减少胞浆中IκBα降解、阻碍NF-κB的过度活化可能是降低炎症因子释放、发挥抗炎作用的重要机制。     

Ghrelin通过核因子-κB抑制肿瘤坏死因子-α诱导的HepG2细胞PAI-1 mRNA表达

目的探讨Ghrelin对肿瘤坏死因子-α(TNF-α)诱导的HepG2细胞纤溶酶原激活物抑制剂-1(PAI-1)mRNA表达的影响及核因子-κB(NF-κB)在其中的作用.方法HepG2细胞培养,加入不同浓度TNF-α,采用逆转录-聚合酶链反应测定(RT-PCR)检测PAI-1 mRNA的水平;免疫印迹法检测胞浆胞核NF-κBp65和胞浆κB抑制蛋白(IκB)的表达.给予Ghrelin预处理1 h后,加入TNF-α检测NF-κKBp65和IκB表达的变化.结果TNF-α(0.1,1,10μg·L-1)浓度依赖地增高HepG2细胞PAI-1 mRNA的表达;Ghrelin组的PAI-1 mRNA表达减少.TNF-α组胞核NF-κBp65表达增加,胞浆IκBα的表达减少;Ghrelin组较TNF-α组胞核NF-κBp65减少,胞浆IκBα的表达增加.结论TNF-α通过NF-κB介导HepG2 PAl-1mRNA的表达;Ghrelin通过抑制NF-κB而抑制TNF-α诱导PAI-1mRNA的表达.     

Tetramethylpyrazine protected photoreceptor cells of rats by modulating nuclear translocation of NF-κB

Aim: To evaluate the effect of tetramethylpyrazine (TMP) injection on retinal damage induced by N-methyl-N-nitrosourea (MNU) in rats and on nuclear factorkappa B (NF-κB) family members. Methods: Female Sprague-Dawley (SD) rats were randomly divided into groups: (i), control group; (ii), model group; and (iii), TMP-injection groups, in which the rats were subdivided into 40 mg/kg, 80 mg/kg and 160 mg/kg groups. Drugs were injected ip into 47-day-old SD rats once a day. At 50 days of age, all rats in the model group and drug groups also received a single ip injection of 60 mg/kg MNU. Rats in group 1 received ip injection of physiological saline. All rats were killed at different times after MNU or physiological saline treatment. The apoptotic index of photoreceptor ceils was calculated by TUNEL labeling; retinal damage was evaluated based on retinal thickness and the expression of NF-nB family members was detected by Western blot. Results: TMP injections, in a dose-dependent manner, suppressed photoreceptor cell apoptosis and decreased its loss in the peripheral retina. As compared with the MNU-treated group, TMP injection at a dose of 160 mg/kg also timedependently upregulated the NF-κB/p65 protein level in the nucleus and downregulated the IκBα protein level in the cytoplasm. However, no protective effect of TMP injection on MNU-induced central retinal damage was found. Conclusion: TMP injection partially protects against MNU-induced retinal damage by upregulating the nuclear translocation of p65 to inhibit photoreceptor cells apoptosis.     

肿瘤坏死因子-α对原代大鼠肾近端小管上皮细胞Toll样受体2表达的调控及机制

目的探讨肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)对原代大鼠肾近端小管上皮细胞Toll样受体2(tolllike receptor2,TLR2)表达的调控以及核因子-κB(nuclearfactor-κB,NF-κB)在其中的作用。方法体外分离与培养原代大鼠肾近端小管上皮细胞,以TNF-α按不同时间给予刺激,Western blot检测TLR2,I-κBα,磷酸化I-κBα(pI-κBα),GAPDH蛋白水平。分别以TNF-α、NF-κB特异性抑制剂Bay11-7082处理25h、Bay11-7082预处理1h后加入TNF-α刺激24h,检测TLR2表达的变化。结果TNF-α刺激6~24h,TLR2高于基线水平;Bay11-7082处理组和Bay11-7082+TNF-α处理组的TLR2蛋白表达水平高于对照组。Bay11-7082+TNF-α处理组与单独TNF-α处理组TLR2蛋白表达水平无差异。TNF-α刺激后5~120min,pI-κBα蛋白水平升高。结论TNF-α促进原代大鼠肾近端小管上皮细胞TLR2蛋白表达,NF-κB对TLR2的表达可能起负调节作用。     

溃疡汤联合雷贝拉唑治疗Hp相关性十二指肠溃疡的疗效及其对患者血清NF-κB与TNF-α的影响

目的 探讨溃疡汤联合雷贝拉唑治疗Hp相关性十二指肠溃疡的疗效及其对患者血清核因子κB (Nuclear factor kappa-B,NF-κB)与肿瘤坏死因子α(Tumor necrosis factor-α,TNF-α)的影响.方法 按照随机数字表法,将96例Hp相关性十二指肠溃疡患者分为对照组与观察组,各48例....     

Inhibitory effect of leflunomide on hepatic fibrosis induced by CCl4 in rats

AIM: To study the effect of leflunomide on CC14-induced hepatic fibrosis in rats. METHODS: Hepatic fibrosis was induced by subcutaneous injection with 50 % CCl4 in Sprague-Dawley rats. The amount of CC14 administered was 1 mg/kg. The alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO) levels in plasma and hydroxyproline (Hyp) contents in liver tissue were assayed by spectrophotometry. The hyaluronic acid (HA) and procollagen III (PC III) were assessed by radioimmunoassay. The transforming growth factor-β1(TGF-β1) in serum was determined by ELISA. The nuclear factor-kappa B (NF-κB) in liver tissue was examined by immunohistochemistry. Liver samples collected after 12 weeks ofCC14 treatment were stained with hematoxy-lin and eosin. RESULTS: Leflunomide (1, 3, and 9 mg/kg) significantly decreased indices of liver and spleen, the serum transaminase (AST, ALT) activities, HA and PC III levels, and Hyp contents in liver tissue in rats of hepatic fibrosis. Histopathological examination showed leflunomide had inhibitory effect on fibrogenesis and formation of pseudolobulus. Furthermore, leflunomide significantly inhibited NF-κB expression in liver tissue, and reduced elevated serum TGF-κB and NO levels in rats of hepatic fibrosis. CONCLUSION: Leflunomide showed inhibitory action on hepatic fibrosis induced by CC14 in rats.     

阻断NF-κB/IκB信号通路对HIBD大鼠脑内炎症因子的影响

目的:建立大鼠缺氧缺血性脑损伤(HIBD)模型,将IκB通过腺病毒载体导入体内,探讨其对HIBD大鼠脑内NF-κB活性和TNF-α、IL-1表达的影响。方法:40只大鼠随机分为5组:I组(正常对照组),II组(HIBD损伤1d组),III组(HIBD损伤7d组),IV组(IκB干预1d组),V组(IκB干预7d组)。Western-blot检测脑细胞NF-κB的表达,了解NF-κB的活性,放免法检测TNF-α、IL-1的表达。结果:经中心静脉途径导入腺病毒转载的IκB基因,可降低HIBD大鼠脑内NF-κB活性,IκB干预1d组和7d组与HIBD损伤1d组和HIBD损伤7d组比较,NF-κB活性明显减少(P<0.05);降低脑内TNF-α、IL-1含量,IκB干预1d组和7d组与HIBD损伤1d组和7d组比较,TNF-α、IL-1含量明显减少(P<0.05)。结论:通过中心静脉途径注入腺病毒转载的IκB基因,直接增加脑内IκB的表达,阻断炎症因子TNF-α的合成,机制可能与其抑制了NF-κB的活性有关。     

晚期糖化终产物受体、核因子-κB双基因反义RNA抑制糖化终产物刺激的炎症因子表达

目的:观察晚期糖化终产物受体(receptor ofadvanced glycation endproducts,RAGE)、核因子-κB(NF-κB)双基因反义RNA对晚期糖化终产物(ad-vanced glycation endproducts,AGEs)刺激ECV304细胞分泌炎症因子的影响。方法:酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)法观察AGEs对ECV304细胞分泌TNF-α和IL-6的影响,应用脂质体将RAGE、NF-κB单/双基因反义RNA转染ECV304,流式细胞仪、筛选低表达RAGE、NF-κB的细胞株,观察RAGE、NF-κB双基因反义RNA对AG-Es刺激ECV304细胞分泌TNF-α和IL-6的影响。结果:AGEs可引起ECV304细胞分泌TNF-α和IL-6增加,诱导作用具有时间和剂量依赖规律。稳定转染筛选出低表达RAGE、NF-κB的细胞株,在AGEs100 mg.L-1诱导下双基因转染细胞ECV-asRAGE-asP65克隆中RAGE、NF-κBp65表达抑制率分别为(62.2±8.7)%及(37.2±7.1)%。AGEs 100 mg.L-1刺激下ECV-asRAGE-asP65克隆分泌TNF-αI、L-6较空载体转染细胞、单基因转染细胞减少更明显(P<0.01)。结论:RAGE、NF-κB双基因反义RNA可抑制AGEs刺激的ECV304细胞的TNF-α和IL-6释放。     

NF-κB的表达情况与冠状动脉病变程度相关性研究

目的研究核因子(NF-κB)通路的表达情况与冠状动脉病变程度相关性。方法 128例行冠脉造影后诊断为冠心病患者(冠心病组),依据血管狭窄程度将其分为轻度病变组(38例)、中度病变组(46例)、重度病变组(44例)。并选择冠状动脉造影正常的50例患者作为对照组。所有患者均检测血清炎症因子白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)及NF-κB,分析IL-6、TNF-α及NF-κB与冠状动脉病变程度的相关性。结果冠心病组IL-1、TNF-α及NF-κB水平明显高于对照组(P<0.05),冠心病各组IL-6、TNF-α、NF-κB水平比较差异有统计学意义(P<0.05),可见IL-6、TNF-α、NF-κB水平随冠状动脉狭窄程度加重而升高。结论血清NF-κB表达水平随冠状动脉病变程度的加重而升高,可作为冠心病治疗的新靶点。