目的探讨苏葶平喘汤对激素抵抗型难治性哮喘小鼠癌蛋白Fos(c-Fos)及血清核转录因子激活蛋白-1(activator protein 1,AP-1)表达的影响,试分析苏葶平喘汤对难治性哮喘的干预机制。方法将50只雌性SPF级BALB/c小鼠随机分为5组,分别为空白组...目的探讨苏葶平喘汤对激素抵抗型难治性哮喘小鼠癌蛋白Fos(c-Fos)及血清核转录因子激活蛋白-1(activator protein 1,AP-1)表达的影响,试分析苏葶平喘汤对难治性哮喘的干预机制。方法将50只雌性SPF级BALB/c小鼠随机分为5组,分别为空白组(A)、模型组(B)、苏葶平喘汤组(C)、地塞米松组(D)和苏葶平喘汤+地塞米松组(E),每组10只。除空白组外,剩余4组均将小鼠建立为激素抵抗型哮喘模型。药物干预结束后进行取材,利用酶联免疫吸附测定(ELISA)法、实时荧光定量聚合酶链式反应(Real-time PCR)分别检测小鼠血清AP-1、肺组织c-Fos的表达水平。结果1.ELISA检测结果显示:模型组小鼠较空白组小鼠血清中AP-1的值明显增高(P<0.05),苏葶平喘汤组、地塞米松组及苏葶平喘汤+地塞米松组较空白组的小鼠血清AP-1值均降低(P<0.05),其中以苏葶平喘汤+地塞米松组的降低最为显著(P<0.05)。2.PCR检测结果显示:模型组与其余各组相比小鼠肺组织的c-FosmRNA表达明显升高(P<0.05),苏葶平喘汤组、地塞米松组和苏葶平喘汤+地塞米松组c-FosmRNA表达下降(P<0.05),地塞米松组、苏葶平喘汤组和苏葶平喘汤+地塞米松组小鼠的肺组织c-FosmRNA表达水平则无明显差异。结论苏葶平喘汤治疗激素抵抗型哮喘可通过影响AP-1及c-Fos的表达水平来实现抑制哮喘炎症反应控制激素抵抗型哮喘症状的作用。展开更多
Magnesium acetyltaurate(MgAT)has been shown to have a protective effect against N-methyl-D-aspartate(NMDA)-induced retinal cell apoptosis.The current study investigated the involvement of nuclear factor kappa-B(NF-κB...Magnesium acetyltaurate(MgAT)has been shown to have a protective effect against N-methyl-D-aspartate(NMDA)-induced retinal cell apoptosis.The current study investigated the involvement of nuclear factor kappa-B(NF-κB),p53 and AP-1 family members(c-Jun/c-Fos)in neuroprotection by MgAT against NMDA-induced retinal damage.In this study,Sprague-Dawley rats were randomized to undergo intravitreal injection of vehicle,NMDA or MgAT as pre-treatment to NMDA.Seven days after injections,retinal ganglion cells survival was detected using retrograde labelling with fluorogold and BRN3A immunostaining.Functional outcome of retinal damage was assessed using electroretinography,and the mechanisms underlying antiapoptotic effect of MgAT were investigated through assessment of retinal gene expression of NF-κB,p53 and AP-1 family members(c-Jun/c-Fos)using reverse transcription-polymerase chain reaction.Retinal phospho-NF-κB,phospho-p53 and AP-1 levels were evaluated using western blot assay.Rat visual functions were evaluated using visual object recognition tests.Both retrograde labelling and BRN3A immunostaining revealed a significant increase in the number of retinal ganglion cells in rats receiving intravitreal injection of MgAT compared with the rats receiving intravitreal injection of NMDA.Electroretinography indicated that pre-treatment with MgAT partially preserved the functional activity of NMDA-exposed retinas.MgAT abolished NMDA-induced increase of retinal phospho-NF-κB,phospho-p53 and AP-1 expression and suppressed NMDA-induced transcriptional activity of NF-κB,p53 and AP-1 family members(c-Jun/c-Fos).Visual object recognition tests showed that MgAT reduced difficulties in recognizing the visual cues(i.e.objects with different shapes)after NMDA exposure,suggesting that visual functions of rats were relatively preserved by pre-treatment with MgAT.In conclusion,pre-treatment with MgAT prevents NMDA induced retinal injury by inhibiting NMDA-induced neuronal apoptosis via downregulation of transcriptional activity of NF-κB,p53 and AP-1-mediated c-Jun/c-Fos.The experiments were approved by the Animal Ethics Committee of Universiti Teknologi MARA(UiTM),Malaysia,UiTM CARE No 118/2015 on December 4,2015 and UiTM CARE No 220/7/2017 on December 8,2017 and Ethics Committee of Belgorod State National Research University,Russia,No 02/20 on January 10,2020.展开更多
BACKGROUND： Nogo-neutralizing antibody IN-1 accelerates axon growth and enhances recovery of spinal cord function by inhibiting growth inhibitory factors. Neurotrophin-3 （NT-3）contributes to regeneration of nerve f...BACKGROUND： Nogo-neutralizing antibody IN-1 accelerates axon growth and enhances recovery of spinal cord function by inhibiting growth inhibitory factors. Neurotrophin-3 （NT-3）contributes to regeneration of nerve fibers in the spinal cord and motor function recovery. The combination of Nogo-neutralizing antibody IN-1 and NT-3 is hypothesized to produce better outcomes and facilitate axonal regeneration by affecting c-Fos and c-Jun protein expression. OBJECTIVE： To investigate the combined effects of Nogo-neutralizing antibody IN-1 and NT-3 on c-Fos and c-Jun protein levels in the injured spinal cord. DESIGN, TIME AND SETTING： A randomized, controlled study was performed at the Laboratory of Neuroanatomy, Xiangya Medical College, Central South University and the Central Laboratory of Third Xiangya Hospital of China from June 2005 to December 2007. MATERIALS： NT-3 （Peprotech, USA） and Nogo-neutralizing antibody IN-1 （Santa Cruz Biotechnology, USA） were used in this study. METHODS： Hemisectioned spinal cord injury models were established by cutting the posterior 2/3 of rat spinal cord, which is equivalent to the T8 level in the human spine. A total of 120 rats were equally and randomly assigned to three groups： model （0.2 μL saline）, IN-1 （0.2 μL IN-1）, and IN-1/NT-3 （0.2 μL IN-1 ＋ 0.2 μL NT-3）. The compounds were separately infused into transection sites on the side of head. MAIN OUTCOME MEASURES： Western blot analysis was employed to measure c-Fos and c-Jun protein expression in the injured spinal cord at 15, 30 minutes, 1,2, 4, 6, 8, and 12 hours following surgery. RESULTS： Following spinal cord injury, c-Fos and c-Jun protein expression were increased and peaked at 4 6 hours. Following injection of IN-1 or the combination of IN-1 and NT-3, c-Fos protein expression was significantly reduced in the injured spinal cord （P 〈 0.05 or P 〈 0.01） （with the exception of the 15 minute time point）. However, c-Jun protein expression was significantly increased （P〈 0.05 or P〈 0.01） （with the exception of the 15 and 30 minute time points）. Combined application of IN-1 and NT-3 resulted in significantly altered protein expression compared to IN-1 alone. CONCLUSION： IN-1 increases c-Jun protein levels and protects the injured spinal cord by inhibiting c-Fos protein levels. Moreover, the effects of IN-1 combined with NT-3 are more significant than with IN-1 alone.展开更多
文摘目的探讨苏葶平喘汤对激素抵抗型难治性哮喘小鼠癌蛋白Fos(c-Fos)及血清核转录因子激活蛋白-1(activator protein 1,AP-1)表达的影响,试分析苏葶平喘汤对难治性哮喘的干预机制。方法将50只雌性SPF级BALB/c小鼠随机分为5组,分别为空白组(A)、模型组(B)、苏葶平喘汤组(C)、地塞米松组(D)和苏葶平喘汤+地塞米松组(E),每组10只。除空白组外,剩余4组均将小鼠建立为激素抵抗型哮喘模型。药物干预结束后进行取材,利用酶联免疫吸附测定(ELISA)法、实时荧光定量聚合酶链式反应(Real-time PCR)分别检测小鼠血清AP-1、肺组织c-Fos的表达水平。结果1.ELISA检测结果显示:模型组小鼠较空白组小鼠血清中AP-1的值明显增高(P<0.05),苏葶平喘汤组、地塞米松组及苏葶平喘汤+地塞米松组较空白组的小鼠血清AP-1值均降低(P<0.05),其中以苏葶平喘汤+地塞米松组的降低最为显著(P<0.05)。2.PCR检测结果显示:模型组与其余各组相比小鼠肺组织的c-FosmRNA表达明显升高(P<0.05),苏葶平喘汤组、地塞米松组和苏葶平喘汤+地塞米松组c-FosmRNA表达下降(P<0.05),地塞米松组、苏葶平喘汤组和苏葶平喘汤+地塞米松组小鼠的肺组织c-FosmRNA表达水平则无明显差异。结论苏葶平喘汤治疗激素抵抗型哮喘可通过影响AP-1及c-Fos的表达水平来实现抑制哮喘炎症反应控制激素抵抗型哮喘症状的作用。
基金supported by Ministry of Higher education,Government of Malaysia,under the grant No.RAGS/2013/UITM/SKK03/2[UiTM file no.600-RMI/RAGS 5/3(103/2013)]the Institut Pengurusan Penyelidikan(RMI),Universiti Teknologi MARA,Malaysia,under the grant 600-IRMI/MyRA 5/3/LESTARI(0088/2016).
文摘Magnesium acetyltaurate(MgAT)has been shown to have a protective effect against N-methyl-D-aspartate(NMDA)-induced retinal cell apoptosis.The current study investigated the involvement of nuclear factor kappa-B(NF-κB),p53 and AP-1 family members(c-Jun/c-Fos)in neuroprotection by MgAT against NMDA-induced retinal damage.In this study,Sprague-Dawley rats were randomized to undergo intravitreal injection of vehicle,NMDA or MgAT as pre-treatment to NMDA.Seven days after injections,retinal ganglion cells survival was detected using retrograde labelling with fluorogold and BRN3A immunostaining.Functional outcome of retinal damage was assessed using electroretinography,and the mechanisms underlying antiapoptotic effect of MgAT were investigated through assessment of retinal gene expression of NF-κB,p53 and AP-1 family members(c-Jun/c-Fos)using reverse transcription-polymerase chain reaction.Retinal phospho-NF-κB,phospho-p53 and AP-1 levels were evaluated using western blot assay.Rat visual functions were evaluated using visual object recognition tests.Both retrograde labelling and BRN3A immunostaining revealed a significant increase in the number of retinal ganglion cells in rats receiving intravitreal injection of MgAT compared with the rats receiving intravitreal injection of NMDA.Electroretinography indicated that pre-treatment with MgAT partially preserved the functional activity of NMDA-exposed retinas.MgAT abolished NMDA-induced increase of retinal phospho-NF-κB,phospho-p53 and AP-1 expression and suppressed NMDA-induced transcriptional activity of NF-κB,p53 and AP-1 family members(c-Jun/c-Fos).Visual object recognition tests showed that MgAT reduced difficulties in recognizing the visual cues(i.e.objects with different shapes)after NMDA exposure,suggesting that visual functions of rats were relatively preserved by pre-treatment with MgAT.In conclusion,pre-treatment with MgAT prevents NMDA induced retinal injury by inhibiting NMDA-induced neuronal apoptosis via downregulation of transcriptional activity of NF-κB,p53 and AP-1-mediated c-Jun/c-Fos.The experiments were approved by the Animal Ethics Committee of Universiti Teknologi MARA(UiTM),Malaysia,UiTM CARE No 118/2015 on December 4,2015 and UiTM CARE No 220/7/2017 on December 8,2017 and Ethics Committee of Belgorod State National Research University,Russia,No 02/20 on January 10,2020.
基金a Grant from Department of Health of Hunan Province of China,No.B2005-076
文摘BACKGROUND： Nogo-neutralizing antibody IN-1 accelerates axon growth and enhances recovery of spinal cord function by inhibiting growth inhibitory factors. Neurotrophin-3 （NT-3）contributes to regeneration of nerve fibers in the spinal cord and motor function recovery. The combination of Nogo-neutralizing antibody IN-1 and NT-3 is hypothesized to produce better outcomes and facilitate axonal regeneration by affecting c-Fos and c-Jun protein expression. OBJECTIVE： To investigate the combined effects of Nogo-neutralizing antibody IN-1 and NT-3 on c-Fos and c-Jun protein levels in the injured spinal cord. DESIGN, TIME AND SETTING： A randomized, controlled study was performed at the Laboratory of Neuroanatomy, Xiangya Medical College, Central South University and the Central Laboratory of Third Xiangya Hospital of China from June 2005 to December 2007. MATERIALS： NT-3 （Peprotech, USA） and Nogo-neutralizing antibody IN-1 （Santa Cruz Biotechnology, USA） were used in this study. METHODS： Hemisectioned spinal cord injury models were established by cutting the posterior 2/3 of rat spinal cord, which is equivalent to the T8 level in the human spine. A total of 120 rats were equally and randomly assigned to three groups： model （0.2 μL saline）, IN-1 （0.2 μL IN-1）, and IN-1/NT-3 （0.2 μL IN-1 ＋ 0.2 μL NT-3）. The compounds were separately infused into transection sites on the side of head. MAIN OUTCOME MEASURES： Western blot analysis was employed to measure c-Fos and c-Jun protein expression in the injured spinal cord at 15, 30 minutes, 1,2, 4, 6, 8, and 12 hours following surgery. RESULTS： Following spinal cord injury, c-Fos and c-Jun protein expression were increased and peaked at 4 6 hours. Following injection of IN-1 or the combination of IN-1 and NT-3, c-Fos protein expression was significantly reduced in the injured spinal cord （P 〈 0.05 or P 〈 0.01） （with the exception of the 15 minute time point）. However, c-Jun protein expression was significantly increased （P〈 0.05 or P〈 0.01） （with the exception of the 15 and 30 minute time points）. Combined application of IN-1 and NT-3 resulted in significantly altered protein expression compared to IN-1 alone. CONCLUSION： IN-1 increases c-Jun protein levels and protects the injured spinal cord by inhibiting c-Fos protein levels. Moreover, the effects of IN-1 combined with NT-3 are more significant than with IN-1 alone.
