Electroacupuncture Delays Cartilage Degeneration by Modulating Nuclear Factor-κB Signaling Pathway

Objective: To illustrate the molecular mechanisms underlying the therapeutic effects of electroacupuncture(EA) on knee osteoarthritis(OA). Methods: Twenty-seven six-month-old New Zealand white rabbits were allocated into three groups in accordance with a random number table: normal group(no surgery-induced OA; without treatment), model group(surgery-induced OA; without treatment) and EA group [surgery-induced OA; received treatment with EA at acupoints Dubi(ST 35) and Neixiyan(EX-LE 5), 30 min twice a day]. After eight consecutive weeks of treatment, the histopathological alterations in cartilage were observed using optical microscopy and transmission electron microscopy, cartilage degeneration was evaluated by modified Mankin's score principles, the synovial fluid concentration of interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) and matrix metalloproteinase-3(MMP-3) were evaluated by enzyme-linked immunosorbent assay, and the protein expression levels of IL-1β, IL-6, TNF-α, MMP-3, IκB kinase-β(IKK-β), nuclear factor of α light polypeptide gene enhancer in B-cells inhibitor α(IκB-α) and nuclear factor-κB(NF-κB) p65 were quantified by Western blot analysis. Results: EA treatment significantly improved cartilage structure arrangement and reduced cellular degeneration. The IL-1β, IL-6, TNF-α and MMP-3 of synovial fluid in the EA-treated group were significantly decreased compared with the model group(all P0.01). Compared with the model group, the IL-1β, IL-6, TNF-α, MMP-3, IKK-β and NF-κB p65 protein expressions in cartilage of EA-treated group were significantly decreased(all P0.01), whereas IκB-α expression was significantly up-regulated(P0.01). Conclusion: EA treatment may delay cartilage degeneration by down-regulating inflammatory factors through NF-κB signaling pathway, which may, in part, explain its clinical efficacy in the treatment of knee OA.     

Activation of NF-κB in bronchial epithelial cells from children with asthma

Objectives To determine whether nuclear factor-κB (NF-κB) is activated in epithelial cells from children with asthma and to understand the role of NF-κB in airway inflammation in asthma. Methods Bronchial mucosa specimens were obtained from 9 children with asthma and 6 control subjects. NF-κB expression in epithelial cells were detected by immunohistochemical examination, and NF-κB-DNA binding was measured by electrophoretic mobility shift assay (EMSA). Results Nuclear expression of NF-κB in epithelial cells was observed in the 9 asthmatic children. NF-κB-DNA binding was found in 4 asthmatic children (EMSA was performed in 6 asthmatic children). In contrast, both nuclear expression and NF-κB-DNA binding were absent in the 6 control subjects. Conclusion These results indicated that NF-κB is activated in epithelial cells from asthmatic children and the NF-κB activation may be the basis for the increased expression of many inflammatory genes and for airway inflammation in asthma.     

Smilax China L. Rhizome Extract Inhibits Nuclear Factor-κB and Induces Apoptosis in Ovarian Cancer Cells

Objective: To study the antitumor effects and associated mechanisms of extract of the Smilax china L. rhizome (SCR) on ovarian cancer cells. Methods: Ovarian cancer cells A2780 were treated with different concentrations of SCR extract (SCRE), and compared with controls. Effects on cell growth were evaluated by cell counting kit-8 (CCK-8) assay; proliferation effects by EdU incorporation assay; cell cycle by propidium iodide staining; apoptosis by annexin V-fluorescein isothiocyanate/propidium iodide; cellular distribution of nuclear factor-κB (NF-κB) by immunofluorescence; protein levels of NF-κB, caspase-3, poly-adenosine diphosphate (ADP)-ribose polymerase (PARP), Bcl-2-associated X protein (Bax), cellular inhibitor of apoptosis (cIAP)-1, anti-X-linked inhibitor of apoptosis protein (XIAP), B-cell lymphoma-extra large (Bcl-XL), B-cell lymphoma-2 (Bcl-2) and AKT by Western blotting; and effects of SCRE combined with cisplatin or adriamycin on A2780 cells by CCK-8 assay. Results: SCRE suppressed A2780 cell proliferation in a dose-dependent manner (P<0.05, P<0.01), arrested cells in G2/M phase and induced apoptosis by activating caspase-3, PARP and Bax. SCRE treatment also correlated with inhibition of NF-κB and downregulation of Bcl-2, Bcl-XL, cIAP-1, XIAP and AKT. SCRE can promote chemosensitivity to cisplatin and adriamycin in A2780 cells (P<0.01). Conclusion: SCR effectively inhibits NF-κB, induces apoptosis and reduces chemoresistance to cisplatin and adriamycin in ovarian cancer cells, which might be its molecular basis for treating ovarian cancer.     

Effect of Triptolide on Expression of Receptor Activator of Nuclear Factor-κB Ligand in Rat Adjuvant Induced Arthritis

Receptor activator of nuclear factor-kappaBligand(RANKL)promotes osteoclast formation,fusion,differentiation and activation.It plays akey role in osteoclast mediated bone erosion inrheumatoid arthritis(RA).Osteoprotegerin(OPG),as a soluble decoy receptor of RANKL,can prevent bone erosion in RA.Our previousstudies have shownthat Triptolide(TP),an activecompoundidentifiedin a traditional Chinese herb--Tripterygium wilfordii Hook F(TWHF),can ef-fectively inhibit bone destruction in ex…     

Extract of Fructus Schisandrae chinensis Inhibits Neuroinflammation Mediator Production from Microglia via NF-κB and MAPK Pathways

Objective: To investigate the anti-neuroinflammation effect of extract of Fructus Schisandrae chinensis(EFSC) on lipopolysaccharide(LPS)-induced BV-2 cells and the possible involved mechanisms. Methods: Primary cortical neurons were isolated from embryonic(E17-18) cortices of Institute of Cancer Research(ICR) mouse fetuses. Primary microglia and astroglia were isolated from the frontal cortices of newborn ICR mouse. Different cells were cultured in specific culture medium. Cells were divided into 5 groups: control group, LPS group(treated with 1 μg/mL LPS only) and EFSC groups(treated with 1 μg/mL LPS and 100, 200 or 400 mg/mL EFSC, respectively). The effect of EFSC on cells viability was tested by methylthiazolyldiphenyltetrazolium bromide(MTT) colorimetric assay. EFSC-mediated inhibition of LPS-induced production of pro-inflammatory mediators, such as nitrite oxide(NO) and interleukin-6(IL-6) were quantified and neuron-protection effect against microglia-mediated inflammation injury was tested by hoechst 33258 apoptosis assay and crystal violet staining assay. The expression of pro-inflammatory marker proteins was evaluated by Western blot analysis or immunofluorescence. Results: EFSC(200 and 400 mg/mL) reduced NO, IL-6, inducible nitric oxide synthase(iN OS) and cyclooxygenase 2(COX-2) expression in LPS-induced BV-2 cel s(P0.01 or P0.05). EFSC(200 and 400 mg/mL) reduced the expression of NO in LPS-induced primary microglia and astroglia(P0.01). In addition, EFSC al eviated cel apoptosis and inflammation injury in neurons exposed to microglia-conditioned medium(P0.01). The mechanistic studies indicated EFSC could suppress nuclear factor(NF)-κB phosphorylation and its nuclear translocation(P0.01). The anti-inflammatory effect of EFSC occurred through suppressed activation of mitogen-activated protein kinase(MAPK) pathway(P0.01 or P0.05). Conclusion: EFSC acted as an anti-inflammatory agent in LPS-induced glia cel s. These effects might be realized through blocking of NF-κB activity and inhibition of MAPK signaling pathways.     

核转录因子κB和糖皮质激素受体在慢性阻塞性肺疾病患者稳定期的相互作用

目的 探讨慢性阻塞性肺疾病(COPD)稳定期外周血和气道黏膜核转录因子κB(NF-κB)和糖皮质激素受体(GCR)的表达.方法 应用Werstern blot方法检测COPD稳定期患者和相匹配的肺功能正常对照组各8例外周血淋巴细胞和支气管黏膜核蛋白核转录因子NF-κB和GCR的表达.结果 COPD患者稳定期外周血淋巴细胞和支气管黏膜核蛋白GCR表达均显著低于对照组(P<0.05,P<0.01),两组支气管黏膜表达显著高于外周血淋巴细胞(P<0.05,P<0.01);COPD患者稳定期外周血淋巴细胞和支气管黏膜核蛋白NF-κB表达均显著高于对照组(P<0.05),两组支气管黏膜表达高于外周血淋巴细胞(P<0.05,P<0.01).结论 COPD患者稳定期与对照组NF-κB和GCR表达差异性提示COPD患者即使在稳定期仍存在慢性炎症,尤其是气道局部炎症更为明显.     

Inhibitory Effects of Parthenolide on the Activity of NF-κB in Multiple Myeloma via Targeting TRAF6

This study examined the mechanism of the inhibitory effect of parthenolide(PTL) on the activity of NF-κB in multiple myeloma(MM). Human multiple myeloma cell line RPMI 8226 cells were treated with or without different concentrations of PTL for various time periods, and then MTT assay was used to detect cell proliferation. Cell cycle and apoptosis were flow cytometrically detected. The level of protein ubiquitination was determined by using immunoprecipitation. Western blotting was employed to measure the level of total protein ubiquitination, the expression of IκB-α in cell plasma and the content of p65 in nucleus. The content of p65 in nucleus before and after PTL treatment was also examined with immunofluorescence. Exposure of RPMI 8226 cells to PTL attenuated the level of ubiquitinated Nemo, increased the expression of IκB-α and reduced the level of p65 in nucleus, finally leading to the decrease of the activity of NF-κB. PTL inhibited cell proliferation, induced apoptosis and blocked cell cycle. Furthermore, the levels of ubiquitinated tumor necrosis factor receptor-associated factor 6(TRAF6) and total proteins were decreased after PTL treatment. By using Autodock software package, we predicted that PTL could bind to TRAF6 directly and tightly. Taken together, our findings suggest that PTL inhibits the activation of NF-κB signaling pathway via directly binding with TRAF6, thereby suppressing MM cell proliferation and inducing apoptosis.     

Role of β2-adrenoceptor-β-arrestin2-nuclear factor-κB signal transduction pathway and intervention effects of oxymatrine in ulcerative colitis

Objective:To investigate the β2-adrenoceptor(β2AR)-β-arrestin2-nuclear factor-κB(NF-κB) signal transduction pathway and the intervention effects of oxymatrine in a rat model of ulcerative colitis.Methods: Forty SD rats were randomly divided into four groups,which included the normal control group,the model group, the mesalazine group and the oxymatrine treatment group,with 10 rats per group.Experimental colitis induced with trinitrobenzene sulfonic acid(TNBS) was established in each group except the normal control group.The rats in the oxymatrine treatment group were treated with intramuscular injection of oxymatrine 63 mg/(kg·d) for 15 days and the rats in the mesalazine group were treated with mesalazine solution 0.5 g/(kg·d) by gastric lavage for 15 days. The rats in the normal control group and model group were treated with 3 mL water by gastric lavage for 15 days. Diarrhea and bloody stool were carefully observed.Histological changes in colonic tissue were examined on day 7 in 2 rats per group that were randomly selected.The expression of β2AR,β-arrestin2 and NF-κB p65 in colon tissue and spleen lymphocytes were detected with immunohistochemistry and Western immunoblotting techniques on day 16 after fasting for 24 h.Six rats died of lavage with 2 each in the normal control,the model group and the mesalazine group;and were not included in the analysis.Results:The rats in the model group suffered from looser stool and bloody purulent stool after modeling.But in the oxymatrine and mesalazine groups,looser stool and bloody purulent stool reduced after treatment.And the colonic wall in the model group was thickened and the colon length shortened.The colon mucosa was congested in multiple areas with edema,erosion,superficial or linear ulcer and scar formation,while the intestinal mucosa injury reduced in the mesalazine and oxymatrine groups(P<0.01).In colonic mucosa and in spleen lymphocytes,compared with the normal control group,the expression of NF-κBp65 were significantly increased(P<0.01) in the model group while the expressions ofβ2AR andβ-arrestin2 were significantly decreased(P<0.01).Compared with the model group,the expression of NF-κBp65 was significantly decreased in the mesalazine group(P<0.01) and oxymatrine treatment group(P<0.01) while the expressions of β2AR and β-arrestin2 were significantly increased(P<0.01).There were no statistically significant differences in the expression of β2AR,β-arrestin2 and NF-κBp65 between the mesalazine group and oxymatrine group(P>0.05).Conclusions:The β2AR-β-arrestin2-NF-κB signal transduction pathway participated in the pathologic course of ulcerative colitis.Oxymatrine attenuated ulcerative colitis through regulating the β2AR-β-arrestin2-NF-κB signal transduction pathway.     

Relationship between Carbachol Hyperstimulation-Induced Pancreatic Acinar Cellular Injury and Trypsinogen or NF-κB Activation in Rats in vitro

In the present study,we determined the rela-tionship between M3cholinergic receptor agonist(carbachol)hypersti mulation-induced pancreaticacinar cellular injury and trypsinogen or NF-κB ac-tivationin ratsin vitro.1MATERIALS AND METHODSCarbachol,collagenaseⅣ,soybean trypsininhibitor(SBTI),BSA,DMEM,HEPES,gluta-mine pyrrolidine dithiocarbamate(PDTC),pefa-bloc,LDH assay kit were obtained from SigmaChemical(USA).Boc-Gln-Ala-Arg-MCA waspurchased from Peptides International(…     

Effects of andrographolide on the activation of mitogen activated protein kinases and nuclear factor-κB in mouse peritoneal macrophage-derived foam cells

Objective:To observe the effect of andrographolide on the activation of mitogen-activated protein kinases(MAPKs) and expression of nuclear factor-κB(NF-k B) in macrophage foam cells.Methods:The mouse peritoneal macrophages were cultured in the media in the presence of oxidized low-density lipoprotein (ox-LDL),ox-LDL+andrographolide,or neither(control).The phosphorylation of MAPK molecules(p38MAPK, JNK,ERK1/2) and the expressions of NK-κB p65 were examined by Western blot.Results:As compared with cells in the control group,the expressions of phospho-p38 and NF-κB p65 were increased in the cells cultured with either ox-LDL or ox-LDL+andrographolide(P<0.01),but attenuated significantly in the presence of ox-LDL+ andrographolide when compared with ox-LDL(P<0.05).The phospho-JNK increased in the presence of either ox-LDL or ox-LDL+andrographolide when compared with control cells(P<0.01),but no significant difference existed between ox-LDL and ox-LDL+andrographolide(P>0.05).The expression of phospho-ERK1/2 was increased in the presence of ox-LDL compared with the control cells(P<0.01),but no significant differences existed between the cells cultured in the presence of ox-LDL+andrographolide and the control medium(P>0.05). Conclusions:Andrographolide could inhibit the activation of ERK1/2,p38MAPK and NK-k B induced by ox-LDL in macrophage foam cells,which might be one of its mechanisms in preventing atherosclerosis.     

Activation of CD40 by soluble recombinant human CD40 ligand inhibits human glioma cells proliferation via nuclear factor-κB signaling pathway

As CD40 transduces activation signals involved in inflammatory and immune disorders,we explored the expression and response to CD40 engagement in human glioma cell lines in this study.The CD40 expression in BT-325 and U251 cells was flow cytometrically detected.The cells were incubated with srhCD40L for 72 h to assess its effects on cell growth in vitro.TNF-α expression was quantified by real-time PCR,and protein expression was analyzed by ELISA.The I-κb mRNA was detected by RT-PCR.I-κB expression decreased after stimulation with 1 μg/mL srhCD40L,but it was upregulated after the cells were pretreated with CD40 antibody.srhCD40L significantly inhibited the proliferation of the CD40+ human glioma cells.The stimulation of CD40+ glioma cells with soluble CD40L (CD154) up-regulated the expression of TNF-α at both mRNA and protein levels.We are led to conclude that CD40L/CD40 could inhibit human glioma cells through I-κb signaling pathway.Interferon-γ can augment CD40 expression and the inhibitory effect of CD40 ligand on cell growth in vitro.These results suggest that srhCD40L may benefit the therapy strategy of glioma.