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Objective:To evaluate the role of spinal ninjurin 2 (NINJ2) in the neuropathic pain (NP) and the relationship with nuclear factor kappa B (NF-κB) signaling pathway in rats.Methods:Thirty-two clean-grade healthy male Sprague-Dawley rats, weighing 230-260 g, aged 7-8 weeks, were divided into 4 groups ( n=8 each) using a random number table method: sham operation group (Sham group), NP group, NP plus NINJ2 interfering virus group (NP+ siRNA group) and NP plus control virus group (NP+ scrRNA group). After intrathecal catheterization, rats in sham group and NP group received normal saline 10 μl, while NP+ NINJ2 siRNA group and NP+ scrRNA group received NINJ2 siRNA 10 μl and scrRNA 10 μl, respectively.NP model was developed via ligation of tibial nerve and common peroneal nerve one week later.Sham group only exposed the sciatic nerve and its branches.The mechanical paw withdrawal threshold (MWT) on the operated side was measured on preoperative days 3 and 1 and postoperative days 1, 3, 5, 7, 10 and 14.The rats were sacrificed at postoperative day 14, and the lumbar enlargement segments of the spinal cord were harvested for determination of the expression of NINJ2 and phosphorylated NF-κB p65 (p-NF-κB p65) (by Western blot) and contents of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 (by enzyme-linked immunosorbent assay). Results:Compared with Sham group, the MWT on the operated side was significantly decreased, the expression of NINJ2 and p-NF-κB p65 in spinal cord was up-regulated, and contents of TNF-α, IL-1β and IL-6 were increased on the postoperative days 3, 5, 7, 10 and 14 in the other three groups ( P<0.05). Compared with NP group, the MWT on the operated side was significantly increased, the expression of NINJ2 and p-NF-κB p65 in spinal cord was down-regulated, and contents of TNF-α, IL-1β and IL-6 were decreased on the postoperative days 3, 5, 7, 10 and 14 in NP+ siRNA group ( P<0.05), and no significant change was found in each parameter mentioned above at different time points in NP+ scrRNA group ( P>0.05). Conclusion:NINJ2 is involved in NP, which is related to activation of NF-κB signaling pathway in rats.
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Chronic pain is a worldwide public health problem in which neuropathic pain (NP) caused by nerve damage is the most common chronic pain.At present,NP lacks effective treatment for any reason.In recent years,increasing studies have shown that neuroimmunity and neuroinflammation play a key role in mediating NP,and nuclear factor-kappa B (NF-κB) plays the most important role in the expression of pain inducers and effectors in the immune and inflammatory systems.NF-κB promotes transcriptional up-regulation by a promoter that links to a variety of inflammatory factors.Therefore,this paper reviews the research status of the role of NF-κB in NP.
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Objective To investigate the effects of penehyclidine hydrochloride on activities of nuclear factor kappa B (NF-kB) and activator protein-1 (AP-1) during actue lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation (HSR) in rats.Methods Thirty male Sprague-Dawley rats,aged 8 weeks,weighing 250-300 g,were randomly assigned into 3 equal groups (n =10 each) using a random number table:sham operation group (group S),blunt chest trauma-HSR group (group THSR) and penehyclidine hydrochloride group (group PHCD).The model of actue lung injury induced by blunt chest trauma-HSR was induced by dropping a 300 g weight onto a precordium in anesthetized rats.Blood was withdrawn via the femoral artery 5 min later until MAP was decreased to 35-45 mmHg within 15 min and maintained at this level for 60 min,followed by resuscitation.In PHCD group,PHCD 2 mg/kg was injected intravenously at 60 min after hemorrhagic shock.At 6 h after the model was established,blood samples were obtained for measurement of concentrations of tumor necrosis factor-alpha (TNF-α) in serum.The lungs were then removed for determination of lung water content,myeloperoxidase (MPO) activaty (by colorimetric assay),NF-κB and AP-1 activaties (using electrophoretic mobility shift assay) in lung tissues,and for microscopic examination of pathologic changes (under light microscope).The left lung was lavaged,and lung permeability index (LPI) was calculated.Results Compared with S group,lung water content,LPI,serum TNF-α level and activites of MPO,NF-κB and AP-1 were significantly increased in THSR and PHCD groups.Compared with THSR group,lung water content,LPI,serum TNF-α concentrations and activites of MPO,NF-κB and AP-1 were significantly decreased in PHCD group.The pathological damage to lung tissues was significantly reduced in PHCD group as compared with THSR group.Conclusion PHCD can inhibit activities of NF-κB and AP-1 in lung tissues,thus mitigating acute lung injury induced by blunt chest trauma-HSR in rats.
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Objective To evaluate expression and significance of TLR4 and NF-κB on inflammatory injure after intracerebral hemorrhage in rats .Methods 60 Sprague Dawley maleness rats were randomly divided into Sham group ,12 h ,24 h ,72 h and 7 d af-ter ICH group(12 s) .The ICH was induced by injection of autologous blood in rats .The behavioral changes were detected by neu-rologic deficit score .The water content of the brain was used to evaluate brain edema changes .Number of TLR4 and NF-κB positive cells by Nissl staining and the expression of protein determined by immunohistochemistry and Western blot .Results After ICH 12 h ,expression of TLR4 and NF-κB positive cells around the hematoma were expressed ,with the extension of the time ,expression was gradually increasing ,and after ICH 72 h the expression of protein were the highest .Cerebral edema and severe neurological damage occurred .Western blot shows the amount of TLR4 expression and NF-κB were in line with the result .Conclusion After in-tracerebral hemorrhage in rat causing inflammatory injure of brain tissue around the hematoma .TLR4 may activate the expression of NF-κB involved in the secondary inflammatory injure after intracerebral hemorrhage in rats .
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Objective To investigate the expression of NF-κBp65 in hippocampus after the XST intervention therapy in the SD rats with global cerebral I/R injury and testify the protective effect of XST after global cerebral I/R injury.Methods 72 healthy SD rats were randomly divided into 3 groups,sham operation(SO) group ( n =24),I/R group( n =24) and XST group( n =24).The model of acute global cerebral ischemia/reperfusion (including:I/R and XST group) injury was produced by means of simple Pulsinelli- brierley's four arteries occlusion method.H.E.staining was performed to detect the number of surviving neurons and TUNEL was used to detect the rate of neurons apoptosis.The expression activation of NF-κB p65 in hippocampus comu-ammonis ( CA1 ) region were examined by immunohistochemical method (SABC).Results The survival pyramidal neurons in the XST group continued to increase,and it was significantly more than the I/R group at each time-point after reperfusion[ (99.23 ±4.22)/mm vs (75.83 ±7.17 )/mm,(80.93 ± 5.36)/mm vs (51.50 ± 8.26 )/mm,(103.24 ± 5.48 )/mm vs (35.67 ± 13.17 )/mm,( 126.22 ± 7.54 )/mm vs (9.83 ± 4.71 )/mm ],the differences were statistically significant ( P ＜0.01 ).The apoptosis rate of pyramidal cell in the XST group at each time-point were more significantly reduced than the I/R group [ ( 8.82 ± 2.71 ) ％ vs ( 22.58 ± 4.68 ) ％.( 19.15 ± 6.23 ) ％ vs (42.68 ± 3.04 ) ％,( 11.82 ± 2.87 ) ％ vs ( 55.51 ± 6.81 ) ％,( 8.44 ± 3.23 ) ％ vs ( 71.69 ± 7.71 ) ％ ],the differences were statistically significant ( P ＜0.01 ).The positive neurons of NF-κBp65 expression in the XST group at different time-points were significantly less than the L/R group[ ( 13.20 ±2.50) vs ( 18.00 ± 1.87),(8.20 ±5.31) vs (41.60±3.65),(6.70±3.36) vs (55.30±5.10),(7.10±3.57) vs (72.80 ±4.71)],the differences were statistically significant ( P ＜ 0.05,P ＜ 0.01 ).Conclusions After global cerebral ischemia/reperfusion,XST could protect the brain from global cerebral ischemia/reperfusion injury by holding up the expression of NF- kappaB p65,and inhibiting neuronal apoptosis,and increasing the number of surviving neurons.Thus,the results of this experiment could provide a powerful and weighty objective indication for XST being used during cerebral resuscitation.
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ObjectiveTo explore the effect and mechanism of Toll-like receptor 4(TLR4) ligand LPS-mediated inhibition hepatitis B virus (HBV) replication in Bewo cells.MethodsFirst of all,2 μg 1.3-fold HBV recombinant vector pcDNA3.1 ( + )-HBV1.3 were transfected into Bewo cells,after 12 h,the cells were treated with LPS for 3 d.To observe the kinetics of IFN-β and TNF-α expression in Bewo cells,the Bewo cells were exposed to TLR4 ligand LPS.And the effect of pyrrolidine dithiocarbamate ( PDTC),an inhibitor of NF-κB,on LPS-induced cytokines was also observed.The HBsAg,HBeAg and HBV DNA level in the culture supernatant were detected by Microparticle Enzyme Immunoassay (MEIA) and fluorescence quantitative PCR,respectively,and the expression of IFN-β,TNF-α,TRIF and MyD88 was detected by ELISA and RT-PCR,respectively.ResultsCompared with control group,LPS could significantly suppress HBV replication in Bewo cells ( P ＜0.01 ),and it could induce the production of TNFα in Bewo cells ( P ＜ 0.05 ),in time-and dose-dependent manners.PDTC strongly inhibited LPS and induced TNF-α production,but had no much effect on IFN-β in Bewo cells ( P ＜ 0.001 ).Compared with control group,the mRNA levels of MyD88 were significantly induced by LPS in the Bewo cells transfected with this recombinant vector( P ＜ 0.001 ).ConclusionsTLR4 ligand LPS could significantly suppress HBV replication by inducing TNF-α production in Bewo cells mainly via the MyD88/ NF-κB signal pathway.