Differential expression of toll-like receptor signaling cascades in LPS-tolerant human peripheral blood mononuclear cells

Pre-exposure to low doses of LPS induces resistance to a lethal challenge, a phenomenon known as endotoxin tolerance. In this study, tolerance was induced in human PBMC by culturing cells with 1 ng/mL LPS for 48 h. Cells were subsequently challenged with 100 ng/mL LPS for 2, 6 and 24 h, and the expression of 84 genes encoding proteins involved in the TLR signaling pathway was evaluated at each time point by PCR array. LPS pretreatment did not modulate the expression of TLR4 and CD14 on the surface of monocytes. A gene was defined as tolerized when LPS pretreatment reversed the effect of LPS challenge on the expression of the gene or as non-tolerized when LPS pretreatment did not reverse the effects of LPS challenge. We observed impaired signal transduction through the NF-κB, JNK, ERK and TRIF pathways, whereas expression of p38 pathway-related genes was preserved in LPS-tolerant cells. These results show a distinct regulation of the TLR pathway cascades during tolerance; this may account for the differential gene expression of some inflammatory mediators, such as up-regulation of IL-10 and COX2 as well as down-regulation of TNF-α and IL-12. Depending on the effect of LPS-induced gene up-regulation or down-regulation, tolerance, as a reversion of such LPS effects, may result in repression or induction of gene expression.     

Intracellular expression and serum levels of eosinophil peroxidase (EPO) and eosinophil cationic protein in asthmatic children

BACKGROUND: Eosinophils are involved in the chronic inflammatory response in asthma and their basic proteins are thought to play a major pathophysiological role in this process. While serum levels of basic proteins have been used to monitor the ongoing allergic disease, little is known about the intracellular expression of these proteins in clinical situations. OBJECTIVE: The aim of the study was to determine the intracellular expression of eosinophil cationic protein (ECP) and eosinophil peroxidase (EPO) in asthmatic children and control subjects and relate it to serum levels of both proteins, lung function tests and immunoglobulin (Ig)E levels. METHODS: Serum ECP and EPO concentrations were determined by immunoassays in 13 asthmatic children (mean age: 9 +/- 1 years, mean FEV1: 92 +/- 10% predicted, geometric mean PC20 histamine 0.5 mg/mL) and 10 age-matched, healthy control subjects. A flow cytometric single cell assay was employed to detect intracellular ECP and EPO in peripheral blood eosinophils. RESULTS: While serum concentrations of both ECP (asthma: median 15.0 microg/L [range 3.6-57.7] vs control: 5.9 microg/L [2.7-9.1]; P = 0.02) and EPO (22.9 microg/L [5.2-82.5] vs 7. 2 microg/L [2.5-12.7]; P = 0.008) were significantly elevated in asthmatics, the intracellular expression of ECP and EPO (measured as mean fluorescence intensity) was decreased (EG1: 55.3 [17.7-120.8] vs 100.3 [46.5-264.4]; P = 0.01; EG2: 80.2 [24.1-135.3] vs 133.7 [32. 1-244.9]; P = 0.04 and EPO: 49.7 [23.1-155.8] vs 94.9 [28.8-115.2]; P = 0.03). In asthmatics there was a significant correlation of FEV1 with intracellular ECP and of bronchial hyperresponsiveness with serum EPO and ECP. Furthermore, total IgE levels were positively correlated with serum EPO only. CONCLUSION: We conclude that in asthmatics the intracellular content of ECP and EPO in peripheral eosinophils is reduced possibly due to degranulation. Epitope masking in activated eosinophils or a shift to early bone marrow-derived progenitors with less granule proteins are further possible explanations.     

Repeated nicotine exposure in rats: effects on memory function, cholinergic markers and nerve growth factor

A decrease in the number of nicotinic-acetylcholine receptors (nAChRs) in the brain is thought to contribute to the cognitive dysfunction associated with diseases as diverse as Alzheimer’s disease and schizophrenia. Interestingly, nicotine and similar compounds have been shown to enhance memory function and increase the expression of nAChRs and therefore, could have a therapeutic role in the aforementioned diseases. Nicotine has also been shown to exert positive effects on certain neurotrophins such as nerve growth factor (NGF), and therefore could play a role beyond mere symptomatic therapy. However, to date, comprehensive studies of nicotine’s effects on the expression of specific acetylcholine (ACh) receptor subtypes, key cholinergic proteins (that are regulated by NGF) such as choline acetyltransferase (ChAT) and the vesicular ACh transporter (VAChT) are lacking. Studies to further investigate the effects of nicotine on NGF especially its high- and low-affinity receptors are also needed. In the present study, male Wistar rats exposed a relatively low dosage of nicotine (0.35 mg/kg every 12 h) for 14 days demonstrated improved memory performance (assessed in two separate water maze testing methods) when compared with controls. Autoradiographic experiments indicated that nicotine increased [³H]-epibatidine, [¹²⁵I]--bungarotoxin and [³H]-AFDX384, but not [³H]-pirenzepine binding sites in several learning- and memory-related brain areas. The expression of ChAT, VAChT, as well as tropomyosin-receptor kinase A (TrkA) NGF receptors and phospho-TrK receptors was increased by nicotine in the hippocampus. No changes were observed in the levels of the NGF peptide or low affinity p75 neurotrophin receptors (p75^NTR), however. These results suggest that repeated exposure to nicotine results in positive effects on central cholinergic markers and memory function, which may be mediated via effects on high-affinity NGF receptors.     

Damage to respiratory epithelium by guinea-pig eosinophils stimulated with IgG-coated Sepharose beads

Background There are no reports of respiratory epithelial damage induced by immuiioglobulin-stimulated eosinophils. Objective We tried to induce damage to respiratory epithelium by guinea-pig (GP) eosinophils stimulated with guinea-pig IgG (GP-lgG)-coated Sepharose 4B beads. Methods GP tracheal epithelium was cultured together with GP eosinophils that had been collected from the peritoneal cavity, purified on a Percoll gradient, and stimulated with GP-lgG-coated beads (GP eosinophils:beads = 20:1). Damage to the epithelium was observed with an inverted microscope. Results After 24 h of culture with three 10⁶ eosinophils. irregularity of the surface of the epithelium, desquamation. shedding of cilia, and abnormal beating of cilia were observed. This damage was first observed after 12 h of incubation, and was more severe at 24 h. No damage was found when beads coated with human serum albumin (HSA) were used. GP eosinophils stimulated with GP-IgG released significantly more EPO than those stimulated with HSA. at 6 and 24 h. Heparin and eatalase partially inhibited the epithelial damage. O₂-production by eosinophils was also enhanced with GP-IgG-coated beads. Conclusion Both granule basic proteins and reactive oxygen radicals may be responsible for epithelial damage, probably via an EPO + H₂O₂+ halide system. These results confirmed that stimulated eosinophils can damage respiratory epithelium.     

Eosinophilic peroxidase deficiency: Identification of a point mutation (D648N) and prediction of structural changes

ABSTRACT Hereditary eosinophil peroxidase (EPO; EC 1.11.1.7) deficiency is a rare abnormality without clinical symptoms characterized by decreased or absent peroxidase activity and decreased volume of the granule matrix in eosinophils. Nearly 100 cases have been reported, but a specific mutation has been reported in only one case. We report the genetic analysis of an EPO-deficient subject and his family. The case was found by automated blood analyzer. Sequencing of the entire coding region of the EPO gene disclosed a novel mutation, a 2060 G-A transition (g. 2060G>A) causing an amino acid change from aspartic acid to asparagine (D648N). Both the son and daughter of the propositus inherited the G-A transition, and in vitro expression experiments suggest this transition is responsible for the deficiency. We then analyzed the location of the affected amino acid within this molecule using a structural model of EPO based on myeloperoxidase (MPO). Asn648 is on the inside of the molecule; changing D to N would cause loss of the electrostatic interaction with Arg146 which is crucial for disulfide bonds of the light chain in the N terminus.     

Eosinophil cationic protein(ECP) and eosinophil protein X (EXP) concentrations in serum and bronchial lavage fluid in asthma. effect of prednisolone treatment.

Background Eoswinophil granule proteins may contribute to hyperresponsiveness in asthma.
Objective To measure eosinophil cationic protein (ECP) and eosinophil protein X (EPX) in sereum and bronchial lavage fluid from 20 asthmatics and 16 control subjects. To asses the effect on these eosinophil proteins of corticosteroid treatment of asthma. To determine ehether serum ECP and EPX measured weekly in a longitudina study for 10 weeks reflected changes in lung function.
Methods Eosinophil granule proteins were measured by radiommunoassy of bronchial wash (BW), bronchoalveolar lavage (BAL) serum.
Results Eosinophils were elevated in BAL (P<0.01) , BW (P<0.01) and blood (P<0.01) from asthmatic compared with control subjects. Eosinophil cationic protein concentration was significantly elevated in BAL (P<0.05) and BW from asthmatics (P<0.01) and EPX was increased in BAL (P<0.05) and BW (P<0.01) . These changes were also reflected in elevated serum ECP (P<0.01) and EPX (P<0.01) concentrations is asthmatic subjects. There was no significant difference between sujects receiving prednisolone and the placebo group, but there was a fall in ECP in BW (P<0.05) and serum (P<0.01) and in EPX in BW (P<0.01) and serum (P<0.01) within the group receiving prednisolone. In the longitudinal study there was only significant difference between ECP values associated with highest and lowest peak expiratory flow rate (PEFR) (P<0.05).
Conclusion These data confirm a role for cosinophil activation in the airway in asthma pathogenesis, and add some support to the hypothesis that corticosteroids may inhibit cosinophil activation in asthma.     

A simple reliable assay for myeloperoxidase activity in mixed neutrophil-eosinophil cell suspensions: application to detection of myeloperoxidase deficiency

Quantitation of myeloperoxidase (MPO) activity by guaiacol peroxidation (GP) assay is profoundly affected by the peroxidase present in eosinophils (EPO) that contaminate the granulocyte suspensions. Inclusion of 3-amino-1,2,4-triazole (AMT) in the GP assay permits quantitation of MPO activity in mixed neutrophil-eosinophil suspension because of the differential inhibition of EPO and MPO by AMT. Results show that: (1) the peroxidase activity of eosinophil-free granulocyte suspensions is not appreciably affected by AMT; (2) in the presence of AMT the peroxidase activities of granulocyte preparations containing different numbers of eosinophils are similar on a neutrophil basis, regardless of the number of eosinophils and correspond with the activity of eosinophil-free granulocyte suspensions; (3) AMT almost completely inhibits the activity of partially purified EPO, only slightly affecting the catalytic activity of partially purified MPO; (4) AMT completely inhibits the residual peroxidase activity of granulocyte suspensions from MPO-deficient subjects contributed by contaminating eosinophils. The GP assay in the presence of AMT was used to study the pattern of hereditary transmission of MPO deficiency. The genealogy derived on the basis of this assay was compatible with an autosomal recessive inheritance, in agreement with previously reported results, while no definite pattern of inheritance could be established by use of the GP assay without AMT. We suggest that the GP assay supplemented with AMT is the method of choice for detection of MPO deficiency, particularly partial deficiency.     

Acetylcholinesterase Activity,Choline Acetyltransferase and Vesicular Acetylcholine Transporter Immunoreactivities in the Rat Adrenal Gland During Postnatal Development

From postnatl‐day‐0 to postnatal‐day‐2, a few acetylcholinesterase (AChE)‐active and choline acetytransferase (ChAT)‐immunoreactive nerve fibers and relatively numerous vesicular acetylcholine transporter (VAChT)‐immunoreactive puncta were observed in the rat adrenal medulla. Despite relatively numerous clear vesicles in the nerve fibers, the synthesis and hydrolysis of acetylcholine may not be fully activated until postnatal‐day‐2. The number of AChE‐active and ChAT‐immunoreactive nerve fibers dramatically increased and that of VAChT‐immunoreactive puncta gradually increased from postnatal‐day‐3 to postnatal‐week‐1. The synthesis and hydrolysis of acetylcholine may be dramatically activated in the nerve fibers of the medulla until postnatal‐week‐1. From postnatal‐week‐2 to postnatal‐week‐3, the number of AChE‐active and the ChAT‐immunoreactive nerve fibers gradually increased and reached the adult levels. The VAChT‐immunoreactive puncta per unit area was maximum number at postnatal‐week‐2. The synthesis and hydrolysis of acetylcholine in the nerve fibers of the medulla may be completed between postnatal‐week‐2 to postnatal‐week‐3. The diameter of the VAChT‐immunoreactive puncta gradually increased from postnatal‐day‐0 with aging. However, the number of the VAChT‐immunoreactive puncta gradually decreased from postnatal‐week‐2 onwards. In electron‐microscopy, the VAChT‐immunoreactive deposits were seen in clusters of clear vesicles, and the diameter of the nerve fibers and the number of clear vesicles at postnatal‐week‐8 increased compared with those at postnatal‐week‐2. The AChE‐active, ChAT‐immunoreactive, and VAChT‐immunoreactive nerve fibers observed around noradrenaline (NA) cells were denser than those around adrenaline (A) cells in the medulla at postnatal‐week‐8. These suggest that the preferential innervation of NA and A cells may cause the differential secretion NA and A. Anat Rec, 292:371–380, 2009. © 2009 Wiley‐Liss, Inc.     

Toll-like receptor 2-mediated sequential activation of MyD88 and MAPKs contributes to lipopolysaccharide-induced sp-a gene expression in human alveolar epithelial cells

Surfactant proteins (SPs) produced by pulmonary epithelial cells participate in the regulation of sepsis-induced acute lung injury. Our previous study has shown that lipopolysaccharide (LPS), a Gram-negative bacterial outer membrane component, can regulate sp-a gene expression in human lung carcinoma type II epithelial A549 cells. This study was further designed to evaluate the signal-transducing mechanisms of LPS-induced sp-a gene expression. Exposure of A549 cells to LPS induced SP-A mRNA and protein production in time-dependent manners. Application of toll-like receptor 2 (TLR2) siRNA into A549 cells decreased the levels of this receptor and simultaneously inhibited LPS-induced SP-A mRNA expression. Sequentially, LPS enhanced phosphorylation of mitogen-activated protein kinase (MEK) 4 and c-Jun NH₂ terminal kinase 1 (JNK1) in time-dependent manners. Application of TLR2 siRNA decreased LPS-enhanced phosphorylation of MEK4 and JNK1. After knocking-down the translation of MyD88 by RNA interference, the LPS-triggered MEK4 phosphorylation was attenuated. Consequently, LPS augmented the translocation of c-Jun from the cytoplasm to nuclei without affecting c-Fos. Pretreatment of A549 cells with SP600125, an inhibitor of JNK1, significantly lowered LPS-induced SP-A mRNA production. Analyses of an electrophoretic mobility shift assay and a reporter gene further showed that LPS increased the transactivation activity of AP-1 in A549 cells. Therefore, the present study demonstrates that LPS can induce sp-a gene expression in human type II epithelial A549 cells through TLR2-mediated sequential activation of MyD88-MEK4-JNK1-AP-1.     

Reduced expression of arrestin beta 2 by graft monocytes during acute rejection of rat kidneys

During acute rejection, numerous pro-inflammatory and cytotoxic monocytes accumulate in the vasculature of experimental renal allografts. Arrestins (ARRBs) are cellular regulators of inflammation, but nothing is known about their expression during rejection. Intravascular mononuclear graft leukocytes were isolated 4 days after kidney transplantation. ARRB1 and ARRB2 mRNA expression was reduced in blood leukocytes from allografts undergoing acute rejection, whereas on the protein level only ARRB2 was changed. Flow cytometry and confocal microscopy revealed ARRB1 and ARRB2 expression by monocytes and T cells, with a selective decrease in ARRB2 expression in monocytes during acute rejection. I-κB directly interacted with ARRB2 and the levels of both proteins strongly correlated. Concomitantly, the mRNA expression of NF-κB targeted genes increased. Our results suggest that activation of blood monocytes in renal isografts is dampened by high ARRB2 levels. During acute rejection, ARRB2 levels are reduced and classical monocyte activation is enabled via NF-κB activation.     

转化生长因子β1对乳鼠心肌细胞转化生长因子结合蛋白2表达的影响

 目的：探讨转化生长因子β1（TGF-β1）在诱导心肌细胞表达转化生长因子结合蛋白2（LTBP2）中的作用及信号传导通路。 方法：培养乳鼠心肌细胞;实时定量聚合酶链式反应（Real-time PCR）、蛋白质印迹和免疫细胞化学方法检测不同时间和不同浓度的TGF-β1对大鼠乳鼠心肌细胞LTBP2基因及蛋白表达的影响;用TGF-β1相关信号通路阻断剂探讨TGF-β1调节LTBP2表达改变的信号传导机制。 结果：LTBP2基因表达随着TGF-β1浓度增加（0、2、5、10 ng/mL）而明显升高,在5 ng/mL时刺激最强（P < 0.05）;5 ng/mL的TGF-β1刺激下心肌细胞内LTBP2基因和蛋白表达的升高呈时间依赖性,均在12 h最高,24 h开始呈下降趋势（P < 0.05或P<0.01）;免疫细胞化学结果显示TGF-β1明显升高LTBP2的表达。信号传导通路研究显示TGF-β1在心肌细胞内主要通过ERK信号通路和PI3K信号通路诱导LTBP2的表达。 结论：TGF-β1在乳鼠心肌细胞内通过ERK信号通路和PI3K信号通路上调LTBP2的表达。     

A novel specificity of anticytoplasmic autoantibodies directed against eosinophil peroxidase.

Vasculitis associated anticytoplasmic autoantibodies (ANCA) are directed against enzymes in the granules of both neutrophils and monocytes. These autoantibodies can be detected by indirect immunofluorescence technique (IIFT) using ethanol-fixed cytospins. We here report the identification of a novel specificity of autoantibodies, present in the sera of eight patients, that reacted only with eosinophils in the IIFT. By immunoprecipitation and ELISA experiments it was shown that the autoantibodies in these sera were directed against eosinophil peroxidase (EPO). There was no apparent influence on initial substrate conversion rate, but reduced plateau levels suggested increased inactivation of the enzyme in the course of the peroxidase reaction. Flow cytometry studies demonstrated the presence of EPO on the surface of primed eosinophils. Anti-EPO sera and purified anti-EPO immunoglobulins significantly increased the release of reactive oxygen species from primed eosinophils. The patients with anti-EPO antibodies suffered from clinically diverse disorders, with more or less generalized manifestations involving the kidneys, blood vessels, lungs and/or joints.     

Migration through basement membrane modulates eosinophil expression of CD44

BACKGROUND: Tissue eosinophils express more membrane receptors and release more mediators than blood eosinophils, suggesting that migration from blood to tissue modulates eosinophil phenotype and functions. OBJECTIVE: We postulated that eosinophil passage through endothelial basement membrane, an important step of eosinophil migration into tissue, may be responsible for some of these changes. METHOD: We previously showed that 5-oxo-6, 8, 11, 14-eicosatetraenoic acid (5-oxo-ETE) in combination with IL-5 promotes eosinophil migration through Matrigel, a mouse tumour cell-derived basement membrane. Using this model, we evaluated the effect of trans-Matrigel migration on purified human blood eosinophil expressions of CD44, CD69 and HLA-DR that either increase or appear on activated eosinophils, and releases of peroxidase (EPO), leukotriene (LT) C(4) and granulocyte-monocyte colony stimulating factor (GM-CSF). RESULTS: IL-5, but not 5-oxo-ETE, increased eosinophil expression of CD44 and CD69. Migration of eosinophils through Matrigel significantly increased CD44 expression level over the one induced by IL-5 (P = 0.0001). Migration through Matrigel did not modify CD69 expression compared with the one obtained in the presence of IL-5 alone; however, incubation of eosinophils on Matrigel decreased IL-5-induced CD69 (P = 0.0001). Trans-Matrigel migration did not modify HLA-DR expression, nor EPO, LTC(4) and GM-CSF releases. CONCLUSION: These data show that in vitro trans-Matrigel migration and Matrigel contact modulate eosinophil membrane receptor expression. Consequently, they suggest that migration through basement membrane mediates changes in cell-surface phenotype observed on activated eosinophils and probably prepares them for interactions with tissue components and cells.     

碱性成纤细胞生长因子对局灶性脑缺血大鼠脑组织核因子-κB表达的影响

目的探讨碱性成纤维细胞生长因子(bFGF)对脑组织缺血再灌注神经元NF-κB的调节作用及机制。方法30只Wista大鼠随机分为Sham组、缺血再灌注组(I/R组)和bFGF组。应用线栓法制作大鼠局灶性脑缺血再灌注模型,大脑中动脉阻塞2h再灌注损伤24h,采用TUNEL法、免疫组化检测海马及皮质内神经元凋亡和NF-κB的表达。结果sham组海马及皮质偶见凋亡细胞,NF-κBp65在细胞核内无表达,在胞浆内极少表达;I/R组海马及皮质神经元凋亡增加,NF-κBp65在细胞内有所表达,皮质及海马NF-κBp65的表达明显高于sham组。bFGF组大脑皮质及海马NF-κBp65的表达较I/R组明显增多。结论bFGF显著减少缺血神经元凋亡,上调脑缺血诱导的NF-κB表达,对脑缺血再灌注海马及皮质神经元的具有保护作用。     

TNF-α and TGF-β1 regulate Syndecan-4 expression in nucleus pulposus cells: role of the mitogen-activated protein kinase and NF-κB pathways

Syndecan-4 is emerging as an important player in cell interaction with the extracellular environment and has been shown to be involved in the progression of intervertebral disc degeneration. However, the mechanism of syndecan-4 regulation by TNF-α and the role of TGF-β1 in regulating syndecan-4 expression remain poorly understood in nucleus pulposus (NP) cells. The aim of this study was to investigate these mechanisms. We exposed NP cells to TNF-α and the gene, protein expression, and promoter activity levels of syndecan-4 were measured by qPCR, western blotting, and the luciferase reporter assay, respectively. The activation of the MAPK and NF-κB pathways was detected using western blot analysis. Syndecan-4 expression in rat NP cells was increased by TNF-α, but this was neither time nor dose dependent in response to TNF-α. ERK1/2, JNK, and NF-κB pathways were activated following TNF-α treatment. Treatment with ERK1/2 and NF-κB inhibitors decreased the up-regulation of syndecan-4 by TNF-α. However, JNK inhibition showed no effect on syndecan-4 expression induced by TNF-α. TNF-α mediated up-regulation of syndecan-4 was antagonized by TGF-β1. This study provided evidence for the differential regulation by MAPK and NF-κB pathways in the over-expression of syndecan-4 promoted by TNF-α in NP cells. Our results demonstrate that TGF-β1 exerts anabolic effects on intervertebral discs by inhibiting the expression of syndecan-4.     

The differential release of eosinophil granule proteins. Studies on patients with acute bacterial and viral infections

Background: Harlier in vitro studies have suggested that the eosinophil may release its granule proteins selectively depending on the stimulus to which the cell is exposed. Objective: The object of the present study was to study the question of selective release in vivo by means of serum measurements of the two eosinophil granule proteins eosinophil cationic protein (ECP) and eosinophil peroxidase (EPO) in acute infections. Methods: Fourty-six subjects with acute infections were studied before treatment, 20 with bacterial infections and 26 with viral infections. Serum ECP, EPO and MPO were measured by specific RIA. Results: In acute bacterial infections ECP, but not EPO. was significantly raised in serum (P < 0.0001) compared with non-infected healthy subjects. In acute bacterial infections ECP was significantly correlated to the levels of the neutrophil marker myeloperoxidase (MPO) (r_s= 0.96, P < 0.0001) but not to EPO. In acute viral infections neither ECP nor EPO were on average raised. However, almost 20% the patients had elevated levels of both proteins. In the viral infections the serum-levels of ECP and EPO were correlated (r_s= 0.63, P < 0.001), but no correlation was found with MPO Conclusion: It is concluded that eosinophils are activated during acute bacterial infections and that this activation results in the preferential mobilisation of ECP. The simultaneous assay of the two eosinophil proteins, ECP and EPO. may give new insight into the role of the eosinophil in disease.     

医用臭氧通过下调神经病理性痛大鼠核因子κB/核因子κB抑制蛋白α/核因子κB抑制蛋白激酶β的表达发挥镇痛效应

目的观察医用臭氧(OZ)对坐骨神经慢性缩窄性损伤(CCI)致神经病理性痛大鼠的镇痛作用及对核因子κB(NF-κB)、核因子κB抑制蛋白α(IκBα)及核因子κB抑制蛋白激酶β(IKKβ)表达水平的影响。方法采用CCI法复制大鼠神经病理性痛动物模型,同时给予不同剂量(0.8、0.4、0.2ml)的OZ予以干预,用Von Frey纤维丝机械刺激触痛仪及冷板测痛仪测定不同剂量OZ对CCI大鼠的机械缩足反射阈值与冷缩足反射阈值的影响;用RT-PCR法和Western blotting法检测不同剂量OZ对CCI大鼠脊髓组织NF-κB p65、IκBα及IKKβmRNA和蛋白表达水平的影响。结果与CCI神经病理性痛模型组比较,OZ 0.8、0.4ml剂量升高CCI大鼠机械缩足反射阈值,降低冷缩足反射阈值(P0.05,P0.01);OZ 0.8、0.4ml剂量可下调CCI大鼠脊髓组织NF-κB p65、IκBα及IKKβmRNA和蛋白表达水平(P0.05,P0.01)。结论 OZ对CCI致神经病理性痛大鼠有镇痛作用,其机制可能与下调NF-κB p65、IκBα及IKKβ的表达有关。