目的探讨心脏组织特异性过表达受体相互作用蛋白140(receptor interacting protein 140,RIP140)对心脏功能及炎症通路的影响。方法大鼠心脏组织多点注射携带RIP140基因的腺病毒载体,使心肌组织过表达RIP140;免疫荧光验证RIP140蛋白在心...目的探讨心脏组织特异性过表达受体相互作用蛋白140(receptor interacting protein 140,RIP140)对心脏功能及炎症通路的影响。方法大鼠心脏组织多点注射携带RIP140基因的腺病毒载体,使心肌组织过表达RIP140;免疫荧光验证RIP140蛋白在心脏组织中的过表达情况;超声心动图与血流动力学参数评估心脏功能;ELISA分析TNF-α、IL-2、IL-1β等炎症因子水平;Western blot分析p65、IκB-α的蛋白水平。结果腺病毒载体介导的外源基因RIP140可成功过表达于心脏组织,诱导心室扩张、左室射血分数下降、心功能受损,促进心肌组织TNF-α、IL-2、IL-1β炎症因子释放,p65蛋白入核、胞质IκB-α蛋白降解。结论腺病毒介导RIP140基因在心脏组织中的特异性过表达损伤心脏功能,激活NF-κB/p65炎症通路,促进炎症因子释放。展开更多
Background：Abnormal neuronal differentiation plays an important role in central nervous system （CNS） development abnormalities such as Down syndrome （DS）,a disorder that results directly from overexpression of ge...Background：Abnormal neuronal differentiation plays an important role in central nervous system （CNS） development abnormalities such as Down syndrome （DS）,a disorder that results directly from overexpression of genes in trisomic cells.Receptor-interacting protein 140 （RIP 140） is significantly upregulated in DS brains,suggesting its involvement in DS CNS development abnormalities.However,the role of RIP140 in neuronal differentiation is still not clear.The current study aimed to investigate the effect of RIP 140 overexpression on the differentiation of neuro-2a （N2a） neuroblastoma cells,in vitro.Methods：Stably RIP 140-overexpressing N2a （N2a-RIP140） cells were used as a neurodevelopmental model,and were constructed by lipofection and overexpression validated by real-time polymerase chain reaction and Western blot.Retinoic acid （RA） was used to stimulate N2a differentiation.Combining the expression of Tuj 1 at the mRNA and protein levels,the percentage of cells baring neurites,and the number of neurites per cell body was semi-quantified to determine the effect of RIP 140 on differentiation of N2a cells.Furthermore,western blot and the ERK1/2 inhibitor U0126 were used to identify the specific signaling pathway by which RIP 140 induces differentiation of N2a cells.Statistical significance of the differences between groups was determined by one-way analysis of variance followed by the Dunnett test.Results：Compared to untransfected N2a cells RIP140 expression in N2a-RIP 140 cells was remarkably upregulated at both the mRNA and protein levels.N2a-RIP 140 cells had a significantly increased percentage of cells baring neurites,and numbers of neurites per cell,as compared to N2a cells,in the absence and presence of RA （P 〈 0.05）.In addition,Tuj l,a neuronal biomarker,was strongly upregulated in N2a-RIP140 cells （P 〈 0.05） and phosphorylated ERK1/2 （p-ERK1/2） levels in N2a-RIP140 cells were dramatically increased,while differentiation was inhibited by the ERK 1/2-specific inhibi展开更多
Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural dif...Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural differentiation remains unclear.We found both RIP140 and ERK1/2 expression increased during neural differentiation of H1 human embryonic stem cells.Moreover,RIP140 negatively correlated with stem cell markers Oct4 and Sox2 during early stages of neural differentiation,and positively correlated with the neural stem cell marker Nestin during later stages.Thus,ERK1/2 signaling may provide the molecular mechanism by which RIP140 takes part in neural differentiation to eventually affect the number of neurons produced.展开更多
文摘目的探讨心脏组织特异性过表达受体相互作用蛋白140(receptor interacting protein 140,RIP140)对心脏功能及炎症通路的影响。方法大鼠心脏组织多点注射携带RIP140基因的腺病毒载体,使心肌组织过表达RIP140;免疫荧光验证RIP140蛋白在心脏组织中的过表达情况;超声心动图与血流动力学参数评估心脏功能;ELISA分析TNF-α、IL-2、IL-1β等炎症因子水平;Western blot分析p65、IκB-α的蛋白水平。结果腺病毒载体介导的外源基因RIP140可成功过表达于心脏组织,诱导心室扩张、左室射血分数下降、心功能受损,促进心肌组织TNF-α、IL-2、IL-1β炎症因子释放,p65蛋白入核、胞质IκB-α蛋白降解。结论腺病毒介导RIP140基因在心脏组织中的特异性过表达损伤心脏功能,激活NF-κB/p65炎症通路,促进炎症因子释放。
基金This research was supported by the grants from the National Natural Science Foundation of China （No. 30872792） and Chief Funds of the National Natural Science Foundation of China （No. 31340024）.
文摘Background：Abnormal neuronal differentiation plays an important role in central nervous system （CNS） development abnormalities such as Down syndrome （DS）,a disorder that results directly from overexpression of genes in trisomic cells.Receptor-interacting protein 140 （RIP 140） is significantly upregulated in DS brains,suggesting its involvement in DS CNS development abnormalities.However,the role of RIP140 in neuronal differentiation is still not clear.The current study aimed to investigate the effect of RIP 140 overexpression on the differentiation of neuro-2a （N2a） neuroblastoma cells,in vitro.Methods：Stably RIP 140-overexpressing N2a （N2a-RIP140） cells were used as a neurodevelopmental model,and were constructed by lipofection and overexpression validated by real-time polymerase chain reaction and Western blot.Retinoic acid （RA） was used to stimulate N2a differentiation.Combining the expression of Tuj 1 at the mRNA and protein levels,the percentage of cells baring neurites,and the number of neurites per cell body was semi-quantified to determine the effect of RIP 140 on differentiation of N2a cells.Furthermore,western blot and the ERK1/2 inhibitor U0126 were used to identify the specific signaling pathway by which RIP 140 induces differentiation of N2a cells.Statistical significance of the differences between groups was determined by one-way analysis of variance followed by the Dunnett test.Results：Compared to untransfected N2a cells RIP140 expression in N2a-RIP 140 cells was remarkably upregulated at both the mRNA and protein levels.N2a-RIP 140 cells had a significantly increased percentage of cells baring neurites,and numbers of neurites per cell,as compared to N2a cells,in the absence and presence of RA （P 〈 0.05）.In addition,Tuj l,a neuronal biomarker,was strongly upregulated in N2a-RIP140 cells （P 〈 0.05） and phosphorylated ERK1/2 （p-ERK1/2） levels in N2a-RIP140 cells were dramatically increased,while differentiation was inhibited by the ERK 1/2-specific inhibi
基金supported by the National Natural Science Foundation of China,No.31340024
文摘Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural differentiation remains unclear.We found both RIP140 and ERK1/2 expression increased during neural differentiation of H1 human embryonic stem cells.Moreover,RIP140 negatively correlated with stem cell markers Oct4 and Sox2 during early stages of neural differentiation,and positively correlated with the neural stem cell marker Nestin during later stages.Thus,ERK1/2 signaling may provide the molecular mechanism by which RIP140 takes part in neural differentiation to eventually affect the number of neurons produced.
