AIM： To investigate the regulation of activin receptor-interacting protein 2 （ARIP2） expression and its possible relationships with collagen type Ⅳ （collagen Ⅳ） in mouse hepatoma cell line Hepal-6 cells. METHOD...AIM： To investigate the regulation of activin receptor-interacting protein 2 （ARIP2） expression and its possible relationships with collagen type Ⅳ （collagen Ⅳ） in mouse hepatoma cell line Hepal-6 cells. METHODS： The ARIP2 mRNA expression kinetics in Hepal-6 cells was detected by RT-PCR, and its regulation factors were analyzed by treatment with signal transduction activators such as phorbol 12-myristate 13-acetate （PMA）, forskolin and A23187. After pcDNA3- ARIP2 was transfected into Hepal-6 cells, the effects of ARIP2 overexpression on activin type Ⅱ receptor （ActRⅡ） and collagen Ⅳ expression were evaluated. RESULTS： The expression levels of ARIP2 mRNA in Hapel-6 cells were elevated in time-dependent manner 12 h after treatment with activin A and endotoxin LPS, but not changed evidently in the early stage of stimulation （2 or 4 h）. The ARIP2 mRNA expression was increased after stimulated with signal transduction activators such as PMA and forskolin in Hepal-6 cells, whereas decreased after treatment with A23187 （25.3% ± 5.7% vs 48.1% ± 3.6%, P 〈 0.01）. ARIP2 overexpression could remarkably suppress the expression of ActRⅡA mRNA in dose-dependent manner, but has no effect on ActRⅡB in Hepal-6 cells induced by activin A. Furthermore, we have found that overexpression of ARIP2 could inhibit collagen Ⅳ mRNA and protein expressions induced by activin A in Hapel-6 cells. CONCLUSION： These findings suggest that ARIP2 expression can be influenced by various factors. ARIP2 may participate in the negative feedback regulation of signal transduction in the late stage by affecting the expression of ActRIIA and play an important role in regulation of development of liver fibrosis induced by activin.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
Regulation of the number of aetivin receptors that are present in the cell membrane plays a key role in the modulation of cellular responses to activin. In order to find the regulators, a novel protein ARIPzip, intera...Regulation of the number of aetivin receptors that are present in the cell membrane plays a key role in the modulation of cellular responses to activin. In order to find the regulators, a novel protein ARIPzip, interacting with activin type II receptors, was searched and identified by using yeast two-hybrid screening. ARIPzip is a splicing variant of ARIP2. This has been discussed previously. ARIPzip can specifically interact with ActR Ⅱ A, and is widely distributed in mouse tissues. Overexpression of ARIPzip can cause the activin-induced transcriptional activities to increase in a dose-dependent manner while the overexpression of ARIV2 can decrease these activities. These data suggest that the C-terminal rezions of ARIP2 and ARIPzip are involved in the regulation of activin signaling.展开更多
Objective： To investigate the proliferation effect and its pathway of activin A on Ovarian epithelial cancer cells line OVCAR-3. Methods： OVCAR-3 cells were cultured in vitro and the membrane receptor ActR II was de...Objective： To investigate the proliferation effect and its pathway of activin A on Ovarian epithelial cancer cells line OVCAR-3. Methods： OVCAR-3 cells were cultured in vitro and the membrane receptor ActR II was detected by immunohistochemical method. The OVCAR-3 cells were cultured with 10 ng/ml activin A for 7 d to observe the effects. Activin A at 5, 10, 15 and 20 ng/ml was used separately to treat the OVCAR-3 cancer cells for 24, 48 and 72 h in order to draw the growth proliferation rate curve measured by MTT method. The expression of protein bcl-2 was detected by western-blot. When OVCAR-3 cells were treated with 10 ng/ml activin A and 5 lag/ml DDP for 24, 48 and 72 h, cell apoptosis could be detected by electron microscopy and flow cytometry （FCM）. Results： Positive expression of ActR II was detected. We also found that the proliferation of OVCAR-3 reached to the climax on the 5th day of culture. The experiments showed that the cells treated with activin A increased quickly and grew faster than those in control group. Moreover, OVCAR-3 cells treated with activin A for 48 h proliferated significantly greater than those treated for 24 h or 72 h （P〈0.01）. bcl-2 protein expression increased expression in activin A treated group than in control group （P〈0.05）. Conclusion： Activin A could increase the proliferation of OVCAR-3 cells which may be through bcl-2 anti-apoptosis pathway.展开更多
Activin receptor-like kinase 1(ALK1)is a transmembrane serine/threonine receptor kinase of the transforming growth factor beta(TGFβ)receptor superfamily.ALK1 is specifically expressed in vascular endothelial cells,an...Activin receptor-like kinase 1(ALK1)is a transmembrane serine/threonine receptor kinase of the transforming growth factor beta(TGFβ)receptor superfamily.ALK1 is specifically expressed in vascular endothelial cells,and its dynamic changes are closely related to the proliferation of endothelial cells,the recruitment of pericytes to blood vessels,and functional differentiation during embryonic vascular development.The pathophysiology of many cerebrovascular diseases is today understood as a disorder of endothelial cell function and an imbalance in the proportion of vascular cells.Indeed,mutations in ALK1 and its co-receptor endoglin are major genetic risk factors for vascular arteriovenous malformation.Many studies have shown that ALK1 is closely related to the development of cerebral aneurysms,arteriovenous malformations,and cerebral atherosclerosis.In this review,we describe the various roles of ALK1 in the regulation of angiogenesis and in the maintenance of cerebral vascular homeostasis,and we discuss its relationship to functional dysregulation in cerebrovascular diseases.This review should provide new perspectives for basic research on cerebrovascular diseases and offer more effective targets and strategies for clinical diagnosis,treatment,and prevention.展开更多
PANoptosis is a newly identified type of regulated cell death that consists of pyroptosis,apoptosis,and nec roptosis,which simultaneously occur during the pathophysiological process of infectious and inflammatory dise...PANoptosis is a newly identified type of regulated cell death that consists of pyroptosis,apoptosis,and nec roptosis,which simultaneously occur during the pathophysiological process of infectious and inflammatory diseases.Although our previous lite rature mining study suggested that PANoptosis might occur in neuronal ischemia/repe rfusion injury,little experimental research has been reported on the existence of PANoptosis.In this study,we used in vivo and in vitro retinal neuronal models of ischemia/repe rfusion injury to investigate whether PAN optosis-like cell death(simultaneous occurrence of pyroptosis,apo ptosis,and necroptosis)exists in retinal neuronal ischemia/repe rfusion injury.Our results showed that ischemia/repe rfusion injury induced changes in morphological features and protein levels that indicate PANoptosis-like cell death in retinal neurons both in vitro and in vivo.Ischemia/repe rfusion inju ry also significantly upregulated caspase-1,caspase-8,and NLRP3 expression,which are important components of the PANoptosome.These results indicate the existence of PANoptosis-like cell death in ischemia/reperfusion injury of retinal neurons and provide preliminary experimental evidence for future study of this new type of regulated cell death.展开更多
目的探讨缺氧缺血性脑病(HIE)新生儿血清生物标志物神经元特异性烯醇酶(NSE)、S-100B、Tau蛋白、激活素A等指标变化及诊断效能。方法选取2020年1月至2022年2月在温州医科大学附属台州医院出生或就诊的75例HIE足月新生儿为研究对象,按照...目的探讨缺氧缺血性脑病(HIE)新生儿血清生物标志物神经元特异性烯醇酶(NSE)、S-100B、Tau蛋白、激活素A等指标变化及诊断效能。方法选取2020年1月至2022年2月在温州医科大学附属台州医院出生或就诊的75例HIE足月新生儿为研究对象,按照HIE严重程度分为轻度HIE组30例,中度HIE组25例,重度HIE组20例。选取同期本院出生的75名健康足月新生儿为对照组。检测并比较4组研究对象血清生物标志物及神经发育指标[包括新生儿神经行为(NBNA)、发育商(DQ)],采用Pearson相关分析血清生物标志物与神经发育指标的相关性,ROC曲线分析各项血清生物标志物单独或联合检测对HIE的诊断效能。结果4组研究对象血清生物标志物及神经发育指标比较,差异均有统计学意义(均P<0.05),其中轻、中、重度HIE组患儿血清NSE、S-100B、Tau蛋白、激活素A水平均明显高于对照组(均P<0.05),且轻度HIE组<中度HIE组<重度HIE组(均P<0.05);生后7 d NBNA评分、生后35周DQ、生后52周DQ均明显低于对照组(均P<0.05),且轻度HIE组>中度HIE组>重度HIE组(均P<0.05)。血清NSE、S-100B、Tau蛋白、激活素A水平与生后7 d NBNA评分均呈负相关(r=-0.779、-0.788、-0.714、-0.800,均P<0.05);与生后35周DQ均呈负相关(r=-0.662、-0.700、-0.626、-0.674,均P<0.05),与生后52周DQ均呈负相关(r=-0.636、-0.684、-0.619、-0.632,均P<0.05)。血清NSE、S-100B、Tau蛋白、激活素A单独检测和4项联合检测诊断HIE的AUC分别为0.923、0.946、0.942、0.939和0.995,4项联合检测的诊断效能最佳。结论血清NSE、S-100B、Tau蛋白、激活素A与HIE新生儿神经发育有关,4项联合检测诊断HIE的效能较高。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 inhibitor U0126.Conclusions：RIP140 overexpression promotes N2a cell neuronal differentiation by activating the ERK1/2 pathway.展开更多
This study aims to demonstrate that blocking the receptor-interacting protein2(Rip2)expression can decrease inflammatory cytokine production by macrophage and protect mice from endotoxin lethality.Murine Rip2 small in...This study aims to demonstrate that blocking the receptor-interacting protein2(Rip2)expression can decrease inflammatory cytokine production by macrophage and protect mice from endotoxin lethality.Murine Rip2 small interfering RNA(siRNA)plasmids were constructed and transfected into macrophage and Rip2 expression was detected with reverse transcription-polymerase chain reaction(RT-PCR)and western blot.Cell proliferation was assayed withMTT.TNF-a concentration was assayed with ELISA and high-mobility group box 1 protein(HMGB1)level with semi-quantitative western blot after lipopolysaccharide(LPS)stimulation.LPS challenge was given after the plasmids were injected into mice and the survival rate was calculated.Rip2 siRNA plasmid could block the mRNA and protein expression of Rip2 and promote cell proliferation.Blocking Rip2 could attenuate LPS-induced TNF-a and HMGB1 production.The HMGB1 expression in the liver decreased to(40.21±11.03)pg/g,and serum TNF-a level decreased to(300.43±59.26)ng/L(P<0.05).The survival rate of mice from endotoxemia was also improved(P<0.05).The results demonstrate that Rip2 siRNA plasmid can block the expression of Rip2,decrease the production of TNF-a and HMGB1 and protect mice from fatal endotoxemia.展开更多
基金Supported by the National Natural Science Foundation of China, No. 30170478 and 30571688Science Projects of Jilin Province of China, No. 20060928-01
文摘AIM： To investigate the regulation of activin receptor-interacting protein 2 （ARIP2） expression and its possible relationships with collagen type Ⅳ （collagen Ⅳ） in mouse hepatoma cell line Hepal-6 cells. METHODS： The ARIP2 mRNA expression kinetics in Hepal-6 cells was detected by RT-PCR, and its regulation factors were analyzed by treatment with signal transduction activators such as phorbol 12-myristate 13-acetate （PMA）, forskolin and A23187. After pcDNA3- ARIP2 was transfected into Hepal-6 cells, the effects of ARIP2 overexpression on activin type Ⅱ receptor （ActRⅡ） and collagen Ⅳ expression were evaluated. RESULTS： The expression levels of ARIP2 mRNA in Hapel-6 cells were elevated in time-dependent manner 12 h after treatment with activin A and endotoxin LPS, but not changed evidently in the early stage of stimulation （2 or 4 h）. The ARIP2 mRNA expression was increased after stimulated with signal transduction activators such as PMA and forskolin in Hepal-6 cells, whereas decreased after treatment with A23187 （25.3% ± 5.7% vs 48.1% ± 3.6%, P 〈 0.01）. ARIP2 overexpression could remarkably suppress the expression of ActRⅡA mRNA in dose-dependent manner, but has no effect on ActRⅡB in Hepal-6 cells induced by activin A. Furthermore, we have found that overexpression of ARIP2 could inhibit collagen Ⅳ mRNA and protein expressions induced by activin A in Hapel-6 cells. CONCLUSION： These findings suggest that ARIP2 expression can be influenced by various factors. ARIP2 may participate in the negative feedback regulation of signal transduction in the late stage by affecting the expression of ActRIIA and play an important role in regulation of development of liver fibrosis induced by activin.
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.
文摘Regulation of the number of aetivin receptors that are present in the cell membrane plays a key role in the modulation of cellular responses to activin. In order to find the regulators, a novel protein ARIPzip, interacting with activin type II receptors, was searched and identified by using yeast two-hybrid screening. ARIPzip is a splicing variant of ARIP2. This has been discussed previously. ARIPzip can specifically interact with ActR Ⅱ A, and is widely distributed in mouse tissues. Overexpression of ARIPzip can cause the activin-induced transcriptional activities to increase in a dose-dependent manner while the overexpression of ARIV2 can decrease these activities. These data suggest that the C-terminal rezions of ARIP2 and ARIPzip are involved in the regulation of activin signaling.
文摘Objective： To investigate the proliferation effect and its pathway of activin A on Ovarian epithelial cancer cells line OVCAR-3. Methods： OVCAR-3 cells were cultured in vitro and the membrane receptor ActR II was detected by immunohistochemical method. The OVCAR-3 cells were cultured with 10 ng/ml activin A for 7 d to observe the effects. Activin A at 5, 10, 15 and 20 ng/ml was used separately to treat the OVCAR-3 cancer cells for 24, 48 and 72 h in order to draw the growth proliferation rate curve measured by MTT method. The expression of protein bcl-2 was detected by western-blot. When OVCAR-3 cells were treated with 10 ng/ml activin A and 5 lag/ml DDP for 24, 48 and 72 h, cell apoptosis could be detected by electron microscopy and flow cytometry （FCM）. Results： Positive expression of ActR II was detected. We also found that the proliferation of OVCAR-3 reached to the climax on the 5th day of culture. The experiments showed that the cells treated with activin A increased quickly and grew faster than those in control group. Moreover, OVCAR-3 cells treated with activin A for 48 h proliferated significantly greater than those treated for 24 h or 72 h （P〈0.01）. bcl-2 protein expression increased expression in activin A treated group than in control group （P〈0.05）. Conclusion： Activin A could increase the proliferation of OVCAR-3 cells which may be through bcl-2 anti-apoptosis pathway.
基金supported by the National Natural Science Foundation of China,No.81801175(to CLT)the Fundamental Research Funds for the Central Universities of China,No.WK9110000044(to CLT)+2 种基金China Scholarship Council,No.201706270155(to CLT)the China Postdoctoral Science Foundation,No.2019M662179(to CLT)the Anhui Province Postdoctoral Science Foundation of China,No.2019B324(to CLT)
文摘Activin receptor-like kinase 1(ALK1)is a transmembrane serine/threonine receptor kinase of the transforming growth factor beta(TGFβ)receptor superfamily.ALK1 is specifically expressed in vascular endothelial cells,and its dynamic changes are closely related to the proliferation of endothelial cells,the recruitment of pericytes to blood vessels,and functional differentiation during embryonic vascular development.The pathophysiology of many cerebrovascular diseases is today understood as a disorder of endothelial cell function and an imbalance in the proportion of vascular cells.Indeed,mutations in ALK1 and its co-receptor endoglin are major genetic risk factors for vascular arteriovenous malformation.Many studies have shown that ALK1 is closely related to the development of cerebral aneurysms,arteriovenous malformations,and cerebral atherosclerosis.In this review,we describe the various roles of ALK1 in the regulation of angiogenesis and in the maintenance of cerebral vascular homeostasis,and we discuss its relationship to functional dysregulation in cerebrovascular diseases.This review should provide new perspectives for basic research on cerebrovascular diseases and offer more effective targets and strategies for clinical diagnosis,treatment,and prevention.
基金supported by the National Natural Science Foundation of China,Nos.81772134,81971891,82172196,81571939(ail to KX)the Key Laboratory of Emergency and Trauma(Hainan Medical University)of Ministry of Education,No.KLET-202108(to KX)+1 种基金the Fundamental Research Funds for the Central Universities of Central South University of China,No.2020zzts218(to WTY)Hunan Provincial Innovation Foundation for Postgraduate of China,No.CX20200116(to WTY)。
文摘PANoptosis is a newly identified type of regulated cell death that consists of pyroptosis,apoptosis,and nec roptosis,which simultaneously occur during the pathophysiological process of infectious and inflammatory diseases.Although our previous lite rature mining study suggested that PANoptosis might occur in neuronal ischemia/repe rfusion injury,little experimental research has been reported on the existence of PANoptosis.In this study,we used in vivo and in vitro retinal neuronal models of ischemia/repe rfusion injury to investigate whether PAN optosis-like cell death(simultaneous occurrence of pyroptosis,apo ptosis,and necroptosis)exists in retinal neuronal ischemia/repe rfusion injury.Our results showed that ischemia/repe rfusion injury induced changes in morphological features and protein levels that indicate PANoptosis-like cell death in retinal neurons both in vitro and in vivo.Ischemia/repe rfusion inju ry also significantly upregulated caspase-1,caspase-8,and NLRP3 expression,which are important components of the PANoptosome.These results indicate the existence of PANoptosis-like cell death in ischemia/reperfusion injury of retinal neurons and provide preliminary experimental evidence for future study of this new type of regulated cell death.
文摘目的探讨缺氧缺血性脑病(HIE)新生儿血清生物标志物神经元特异性烯醇酶(NSE)、S-100B、Tau蛋白、激活素A等指标变化及诊断效能。方法选取2020年1月至2022年2月在温州医科大学附属台州医院出生或就诊的75例HIE足月新生儿为研究对象,按照HIE严重程度分为轻度HIE组30例,中度HIE组25例,重度HIE组20例。选取同期本院出生的75名健康足月新生儿为对照组。检测并比较4组研究对象血清生物标志物及神经发育指标[包括新生儿神经行为(NBNA)、发育商(DQ)],采用Pearson相关分析血清生物标志物与神经发育指标的相关性,ROC曲线分析各项血清生物标志物单独或联合检测对HIE的诊断效能。结果4组研究对象血清生物标志物及神经发育指标比较,差异均有统计学意义(均P<0.05),其中轻、中、重度HIE组患儿血清NSE、S-100B、Tau蛋白、激活素A水平均明显高于对照组(均P<0.05),且轻度HIE组<中度HIE组<重度HIE组(均P<0.05);生后7 d NBNA评分、生后35周DQ、生后52周DQ均明显低于对照组(均P<0.05),且轻度HIE组>中度HIE组>重度HIE组(均P<0.05)。血清NSE、S-100B、Tau蛋白、激活素A水平与生后7 d NBNA评分均呈负相关(r=-0.779、-0.788、-0.714、-0.800,均P<0.05);与生后35周DQ均呈负相关(r=-0.662、-0.700、-0.626、-0.674,均P<0.05),与生后52周DQ均呈负相关(r=-0.636、-0.684、-0.619、-0.632,均P<0.05)。血清NSE、S-100B、Tau蛋白、激活素A单独检测和4项联合检测诊断HIE的AUC分别为0.923、0.946、0.942、0.939和0.995,4项联合检测的诊断效能最佳。结论血清NSE、S-100B、Tau蛋白、激活素A与HIE新生儿神经发育有关,4项联合检测诊断HIE的效能较高。
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 inhibitor U0126.Conclusions：RIP140 overexpression promotes N2a cell neuronal differentiation by activating the ERK1/2 pathway.
文摘This study aims to demonstrate that blocking the receptor-interacting protein2(Rip2)expression can decrease inflammatory cytokine production by macrophage and protect mice from endotoxin lethality.Murine Rip2 small interfering RNA(siRNA)plasmids were constructed and transfected into macrophage and Rip2 expression was detected with reverse transcription-polymerase chain reaction(RT-PCR)and western blot.Cell proliferation was assayed withMTT.TNF-a concentration was assayed with ELISA and high-mobility group box 1 protein(HMGB1)level with semi-quantitative western blot after lipopolysaccharide(LPS)stimulation.LPS challenge was given after the plasmids were injected into mice and the survival rate was calculated.Rip2 siRNA plasmid could block the mRNA and protein expression of Rip2 and promote cell proliferation.Blocking Rip2 could attenuate LPS-induced TNF-a and HMGB1 production.The HMGB1 expression in the liver decreased to(40.21±11.03)pg/g,and serum TNF-a level decreased to(300.43±59.26)ng/L(P<0.05).The survival rate of mice from endotoxemia was also improved(P<0.05).The results demonstrate that Rip2 siRNA plasmid can block the expression of Rip2,decrease the production of TNF-a and HMGB1 and protect mice from fatal endotoxemia.
