夏天无注射液参与脑缺血再灌注大鼠神经保护的NF-κB介导机制
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摘要
目的:探讨夏天无注射液参与脑缺血再灌注大鼠神经保护的NF-κB介导机制。方法:随机将雄性SD大鼠分为假手术组、模型组、夏天无注射液1.0 ml/kg的低剂量组、2.5 ml/kg的中剂量组、5 ml/kg的高剂量组及NF-κB抑制剂(BAY11-7082)组。脑缺血再灌注24 h后,2,3,5氯化三苯基四唑(TTC)染色检测各组大鼠脑梗死重量百分比;免疫组织化学技术及Western blot技术检测各组的磷酸化NF-κB表达变化。结果:TTC发现:不同剂量的夏天无注射液及BAY可以减轻脑梗死的总量,其中,高剂量组、中剂量组及抑制剂组之间效果无差异,但三者效果均较低剂量组明显;免疫组化发现:主要在模型组中的海马CA1区细胞核中发现磷酸化NF-κB表达,而CA3区的表达量较少。Western blot检测发现:不同剂量的夏天无注射液可以模拟BAY11-7082的作用,降低磷酸化NF-κB蛋白的表达量。结论:夏天无注射液可能减轻大鼠脑缺血再灌注后梗死程度,其机制可能是通过抑制NF-κB的过度磷酸化。
Objective: To investigate themechanism on NF-κB mediates the injection coryadlis decumbens pers( ICDP) participated in neuroprotection after ischemia reperfusion of rats. Methods: The SD rats were rando mly divided into several groups as follows:Sham operation group,Model group,1. 0 ml / kg ICDP group( Low-dose,ICDP-L),2. 5 ml / kg ICDP group( Middle-dose,ICDP-M),5ml / kg ICDP group( High-dose,ICDP-H),and NF-κB inhibitor group( BAY11-7082). 24 h after anesthetize,the volume of infarct sections in different groups were detected by TCC staining,and the phosphorylated NF-κB expression in rats brain was observed by immunohistochemistry and Western blot. Results: The TTC staining showed that different concentration of ICDP and BAY11-7082 could reduce the brain infarction volume significantly. There was no significant different effect among the ICDP-H group,ICDP-M group and inhibitor group,however,the effect in these three groups was more effective than that in the ICDP-M group. In addition,the results of immunohistochemistry indicated that phosphorylated NF-κB p65 expressed in brain tissue located mainly at the nucleus neuronal cells in the CA1 region of hippocampusin model rats,and the expression of phosphorylated NF-κB were significantly reduced in ICDP groups and BAY11-7082 group. Conclusion: The ICDP can reduce brain infarct volume after ischemia reperfusion of rats. The neuralprotection mechanism of ICDP may relative toinhibits thehyperphosphorylation of NF-κB.
Objective: To investigate themechanism on NF-κB mediates the injection coryadlis decumbens pers( ICDP) participated in neuroprotection after ischemia reperfusion of rats. Methods: The SD rats were rando mly divided into several groups as follows:Sham operation group,Model group,1. 0 ml / kg ICDP group( Low-dose,ICDP-L),2. 5 ml / kg ICDP group( Middle-dose,ICDP-M),5ml / kg ICDP group( High-dose,ICDP-H),and NF-κB inhibitor group( BAY11-7082). 24 h after anesthetize,the volume of infarct sections in different groups were detected by TCC staining,and the phosphorylated NF-κB expression in rats brain was observed by immunohistochemistry and Western blot. Results: The TTC staining showed that different concentration of ICDP and BAY11-7082 could reduce the brain infarction volume significantly. There was no significant different effect among the ICDP-H group,ICDP-M group and inhibitor group,however,the effect in these three groups was more effective than that in the ICDP-M group. In addition,the results of immunohistochemistry indicated that phosphorylated NF-κB p65 expressed in brain tissue located mainly at the nucleus neuronal cells in the CA1 region of hippocampusin model rats,and the expression of phosphorylated NF-κB were significantly reduced in ICDP groups and BAY11-7082 group. Conclusion: The ICDP can reduce brain infarct volume after ischemia reperfusion of rats. The neuralprotection mechanism of ICDP may relative toinhibits thehyperphosphorylation of NF-κB.
引文
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[18]Meng Z,Lou S,Tan J,et al.Nuclear factor-kappa b inhibition can enhance apoptosis of differentiated thyroid cancer cells induced by131Ⅰ[J].Plo S One,2012,7(3):e33597.
[2]Ahmad M,Graham SH.Inflammation after stroke:Mechanisms and therapeutic approaches[J].Translational Stroke Res,2010,1(2):74-84.
[3]Candelario-Jalil E.Injury and repair mechanisms in ischemic stroke:Considerations for the development of novel neuroth-erapeutics[J].Curr Opin Investig Drugs(London,England:2000),2009,10(7):644-654.
[4]Macrez R,Ali C,Toutirais O,et al.Stroke and the immune system:From pathophysiology to new therapeutic strategies[J].Lancet Neurol,2011,10(5):471-480.
[5]van der Kooij MA,Nijboer CH,Ohl F,et al.Nf-kappab inhibition after neonatal cerebral hypoxia-ischemia improves long-term motor and cognitive outcome in rats[J].Neurobiol Disease,2010,38(2):266-272.
[6]Fan H,Li L,Zhang X,et al.Oxymatrine downregulates tlr4,tlr2,myd88,and nf-kappab and protects rat brains against focal ischemia[J].Mediators of inflammation,2009,2009:704-706.
[7]Dabek J,Kulach A,Gasior Z.Nuclear factor kappa-light-chain-enhancer of activated b cells(nf-kappab):A new potential therapeutic target in atherosclerosis?[J].Pharmacological reports,2010,62(5):778-783.
[8]姚丽梅,刘瑶,段启.注射用夏天无总碱对大鼠局灶性脑缺血的影响[J].中成药,2011,33(5):872-874.
[9]Alvira CM,Abate A,Yang G,et al.Nuclear factor-kappab activation in neonatal mouse lung protects against lipopolysacc-haride-induced inflammation[J].Am J Respiratory Critical Care Med,2007,175(8):805-815.
[10]Zador Z,Benyo Z,Lacza Z,et al.[Neuroprotection in brain ischemia-doubts and hopes][J].Ideggyogyaszati szemle,2004,57(3-4):81-93.
[11]Fagan SC,Hess DC,Machado LS,et al.Tactics for vascular protection after acute ischemic stroke[J].Pharmacotherapy,2005,25(3):387-395.
[12]Harari OA,Liao JK.Nf-kappab and innate immunity in ischemic stroke[J].Annals NY Acad Sci,2010,1207:32-40.
[13]Tamatani M,Che YH,Matsuzaki H,et al.Tumor necrosis factor induces bcl-2 and bcl-x expression through nfkappab activation in primary hippocampal neurons[J].J biological chem,1999,274(13):8531-8538.
[14]Saji C,Higashi C,Niinaka Y,et al.Proteasome inhibitors induce apoptosis and reduce viral replication in primary effusion lymphoma cells[J].Biochemical Biophysical Res Communications,2011,415(4):573-578.
[15]Spinelli SL,Casey AE,Pollock SJ,et al.Platelets and megakaryocytes contain functional nuclear factor-kappab[J].Arterioscle Thromb Vasc Biol,2010,30(3):591-598.
[16]Pfeffer LM.The role of nuclear factor kappab in the interferon response[J].J Interferon Cytokine Res,2011,31(7):553-559.
[17]Ma W,Dumont Y,Vercauteren F,et al.Lipopolysaccharide induces calcitonin gene-related peptide in the raw264.7 macrophage cell line[J].Immunology,2010,130(3):399-409.
[18]Meng Z,Lou S,Tan J,et al.Nuclear factor-kappa b inhibition can enhance apoptosis of differentiated thyroid cancer cells induced by131Ⅰ[J].Plo S One,2012,7(3):e33597.