内质网是真核细胞中重要的细胞器之一,与维持细胞稳态关系密切。当缺乏葡萄糖、缺氧、体内钙平衡紊乱或者发生氧化应激时,会引起细胞内未折叠蛋白或错误折叠蛋白的积累,导致内质网应激。帕金森病是一种慢性进行性脑变性疾病,典型的病理...内质网是真核细胞中重要的细胞器之一,与维持细胞稳态关系密切。当缺乏葡萄糖、缺氧、体内钙平衡紊乱或者发生氧化应激时,会引起细胞内未折叠蛋白或错误折叠蛋白的积累,导致内质网应激。帕金森病是一种慢性进行性脑变性疾病,典型的病理变化是黑质纹状体多巴胺能神经细胞变性丢失导致的多巴胺神经递质缺乏。目前对帕金森病的治疗多为缓解症状,但不能阻止疾病的进展。通过对内质网应激中的信号通路的研究发现:在帕金森病的发病过程中,多巴胺能神经元的选择性死亡与内质网应激有关。内质网应激过程中的中心调节因子:葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)及其下游ATF4-CHOP-Puma信号通路与帕金森病的发病过程有密切的联系,本文对GRP78及其下游ATF4-CHOP-Puma信号通路近些年来的研究进展进行综述,以期为帕金森病的治疗提供新的靶点和思路。展开更多
Objective:To examine whether exposure of mouse neuronal cells to radiofrequency fields used in mobile communication devices can induce stress in endoplasmic reticulum(ER)and activate unfolded protein response(UPR).Met...Objective:To examine whether exposure of mouse neuronal cells to radiofrequency fields used in mobile communication devices can induce stress in endoplasmic reticulum(ER)and activate unfolded protein response(UPR).Methods:HT22 mouse hippocampus neuronal cells were exposed to continuous wave 900 MHz radiofrequency fields(RF)at 120μW/cm2 power intensity for 4 h/d for 5 consecutive days.The positive control cells were irradiated with 4 Gy of 60Coγ-rays at a dose rate of 0.5 Gy/min(GR).Twenty-four hours after the last exposure,cells were collected,and the expressions of sensor transmembrane proteins were detected using Western blot analysis.Results:The expression levels of Ire1,PERK,p-IRE1 and p-PERK,GRP78 and CHOP proteins were detected.There were no statistically significant differences in the expression levels of IRE1 and PERK proteins in control(CT),sham(SH)-,RF-and GR-exposed cells(P<0.05).The phosphorylated protein levels of p-IRE1 and p-PERK were significantly increased in cells exposed to RF and GR(P<0.05).The expression levels of GRP78 and CHOP were significantly increased in RF-and GR-exposed cells compared to CT and SH-exposed cells(P<0.05).Cells treated with 1μg/ml TM for 24 h showed significantly increased expression levels of GRP78 and CHOP proteins compared to controls(P<0.05).In the presence of 2 mmol/L PBA,TM-induced increased levels of GRP78 and CHOP proteins were reduced(P<0.05).Conclusions:The exposure of non-ionizing 900 MHz RF was able to cause stress in HT22 mouse neuronal cells and activated UPR in ER.Since UPR plays an important role in both cell survival(when UPR is mitigated)and apoptosis/death(under unresolvable stress conditions),further studies are required to determine the fate of the cells exposed to RF.展开更多
本研究探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对自然衰老大鼠内质网应激相关蛋白的调节作用以及对神经细胞凋亡的干预作用。雄性SD大鼠随机分为6月龄组、24月龄组、SPJ低剂量组(10mg/kg)和高剂量组(30mg/kg)。Westernblo...本研究探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对自然衰老大鼠内质网应激相关蛋白的调节作用以及对神经细胞凋亡的干预作用。雄性SD大鼠随机分为6月龄组、24月龄组、SPJ低剂量组(10mg/kg)和高剂量组(30mg/kg)。Westernblot法检测大鼠皮层和海马内质网应激相关蛋白(GRP78、p-PERK、p-EIF2α、ATF4、ATF6、p-IRE1、XBP1)以及凋亡相关蛋白(p-JNK、CHOP、Bcl-2、Bax)表达变化。结果显示,与6月龄组相比,24月龄组皮层/海马组织中GRP78、p-PERK、p-EIF2α、ATF4、ATF6、p-IRE1、XBP1和Bcl-2/Bax蛋白表达分别下降了80%/59%、82%/78%、79%/77%、94%/74%、72%/66%、78%/81%、82%/71%和90%/92%,p-JNK/JNK、CHOP蛋白表达分别上升了1/1.72、3.33/1.33倍。给予不同剂量竹节参总皂苷干预后,均能明显上调内质网应激相关蛋白和Bcl-2/Bax的表达水平,下调促凋亡蛋白表达水平。研究结果表明,竹节参总皂苷能通过上调内质网应激相关蛋白以及抗凋亡蛋白的表达水平,下调促凋亡蛋白的表达,从而维持衰老大鼠脑组织的内质网稳态,减少神经细胞凋亡。展开更多
目的探讨CAAT增强子结合蛋白同源蛋白(CHOP)、海马葡萄糖调节蛋白78(GRP78)和caspase-12在蛛网膜下腔出血(SAH)后的表达及其在早期脑损伤(EBI)中的作用。方法选取健康雄性SD大鼠78只,采用随机数字表法将其分为空白对照组(6只)、假手术组...目的探讨CAAT增强子结合蛋白同源蛋白(CHOP)、海马葡萄糖调节蛋白78(GRP78)和caspase-12在蛛网膜下腔出血(SAH)后的表达及其在早期脑损伤(EBI)中的作用。方法选取健康雄性SD大鼠78只,采用随机数字表法将其分为空白对照组(6只)、假手术组(36只)与SAH组(36只)。将假手术组和SAH组分别于1、6、12、24、48、72h,采用简单随机抽样法选取6只大鼠,神经功能缺陷评分后处死。比较各组不同时间点光镜及电镜下大鼠海马神经元形态改变、CHOP、GRP78、caspase-12相对表达水平、海马神经元凋亡细胞水平、脑水肿严重程度及神经功能缺陷评分情况。结果通过Western blot法检测CHOP、GRP78和caspase-12的表达,SAH组与空白对照组、假手术组在12、24、48、72h比较,差异均有统计学意义(P<0.05),且在SAH后24h达到峰值。TUNEL法检测大鼠海马区神经元凋亡显示,SAH组与对照组、假手术组在6、12、24、48、72h比较,差异均有统计学意义(P<0.05),在电镜下,SAH后24h节点可见明显的神经元凋亡。SAH组12、24、48、72h的神经功能缺陷评分及脑水肿程度明显重于空白对照组及假手术组,在24h达高峰,差异均有统计学意义(P<0.05)。SAH组大鼠的CHOP、GRP78和caspase-12 Western blot法检测结果与脑水肿严重程度及神经元凋亡结果之间呈极显著正相关,与神经功能缺陷评分呈极显著负相关(P<0.05)。结论 SAH后72h内确实造成了EBI,同时发生了内质网应激,内质网应激与EBI严重程度之间均呈正相关,揭示内质网应激在SAH后EBI的病理过程中发挥了重要作用。展开更多
【目的】探讨四逆汤对阿霉素诱导的慢性心力衰竭(CHF)大鼠内质网应激的影响。【方法】从90只大鼠中随机选取15只作为正常组,其余大鼠腹腔注射阿霉素构建CHF模型,同时,正常组给予腹腔注射生理盐水。造模结束后,正常组大鼠全部存活,造模...【目的】探讨四逆汤对阿霉素诱导的慢性心力衰竭(CHF)大鼠内质网应激的影响。【方法】从90只大鼠中随机选取15只作为正常组,其余大鼠腹腔注射阿霉素构建CHF模型,同时,正常组给予腹腔注射生理盐水。造模结束后,正常组大鼠全部存活,造模大鼠存活60只。再将60只CHF大鼠随机分为5组,即模型组,四逆汤低、中、高剂量组,曲美他嗪组,每组12只。四逆汤低、中、高剂量组分别给予1.4、4.2、12.6(生药)g·kg^(-1)·d^(-1)灌胃,曲美他嗪组给予曲美他嗪10 mg·kg^(-1)·d^(-1)灌胃,正常组和模型组给予相同体积的生理盐水灌胃。连续给药4周。采用小动物超声测量左心室舒张末期直径(LVEDD)、左心室射血分数(LVEF)和左心室缩短分数(LVFS)变化;采用酶联免疫吸附分析(ELISA)检测血浆脑钠肽(BNP)和血管紧张素Ⅱ(AngⅡ)水平;采用实时荧光定量聚合酶链反应(PCR)技术检测心肌组织葡萄糖调节蛋白78(GRP78)、RNA依赖的蛋白激酶样内质网激酶(PERK)和活化转录因子4(ATF4)mRNA表达。【结果】与正常组比较,模型组大鼠LVEDD显著增大,LVEF、LVFS均显著降低,血浆BNP和AngⅡ含量升高,心肌组织GRP78、PERK及ATF4 m RNA表达量升高(P <0.01);与模型组比较,四逆汤低、中、高剂量组及曲美他嗪组LVEDD降低,LVEF、LVFS均显著增加,血浆BNP和AngⅡ含量减少,心肌组织GRP78、PERK及ATF4 m RNA表达量降低,且呈剂量依赖性,其中,四逆汤高剂量组及曲美他嗪组差异均有统计学意义(P <0.05或P <0.01)。【结论】四逆汤可改善慢性心力衰竭大鼠模型心脏功能,其机制可能与抑制PERK/ATF4信号通路从而减轻内质网应激有关。展开更多
Class A scavenger receptor （SR-A） plays an important role in macrophage adhesion. However, the underlying mechanism remains unclear. We previously found that 78 kDa glucose-regulated protein （GRP78） inhibited SR- ...Class A scavenger receptor （SR-A） plays an important role in macrophage adhesion. However, the underlying mechanism remains unclear. We previously found that 78 kDa glucose-regulated protein （GRP78） inhibited SR- A-mediated ligand internalization into macrophage by binding to SR-A. The aim of the study was to investigate whether GRP78 could regulate SR-A-mediated cell adhesion. We demonstrated that GRP78 bound directly to SR-A by fluorescence resonance energy transfer （FRET） assay. Overexpression of GRP78 inhibited macrophage adhesion via SR-A. These results suggest that GRP78 may act as an inhibitor of macrophage adhesion via SR-A.展开更多
文摘内质网是真核细胞中重要的细胞器之一,与维持细胞稳态关系密切。当缺乏葡萄糖、缺氧、体内钙平衡紊乱或者发生氧化应激时,会引起细胞内未折叠蛋白或错误折叠蛋白的积累,导致内质网应激。帕金森病是一种慢性进行性脑变性疾病,典型的病理变化是黑质纹状体多巴胺能神经细胞变性丢失导致的多巴胺神经递质缺乏。目前对帕金森病的治疗多为缓解症状,但不能阻止疾病的进展。通过对内质网应激中的信号通路的研究发现:在帕金森病的发病过程中,多巴胺能神经元的选择性死亡与内质网应激有关。内质网应激过程中的中心调节因子:葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)及其下游ATF4-CHOP-Puma信号通路与帕金森病的发病过程有密切的联系,本文对GRP78及其下游ATF4-CHOP-Puma信号通路近些年来的研究进展进行综述,以期为帕金森病的治疗提供新的靶点和思路。
基金This research is supported by funding from the National Natural Science Foundation of China(Grant No.81373025).
文摘Objective:To examine whether exposure of mouse neuronal cells to radiofrequency fields used in mobile communication devices can induce stress in endoplasmic reticulum(ER)and activate unfolded protein response(UPR).Methods:HT22 mouse hippocampus neuronal cells were exposed to continuous wave 900 MHz radiofrequency fields(RF)at 120μW/cm2 power intensity for 4 h/d for 5 consecutive days.The positive control cells were irradiated with 4 Gy of 60Coγ-rays at a dose rate of 0.5 Gy/min(GR).Twenty-four hours after the last exposure,cells were collected,and the expressions of sensor transmembrane proteins were detected using Western blot analysis.Results:The expression levels of Ire1,PERK,p-IRE1 and p-PERK,GRP78 and CHOP proteins were detected.There were no statistically significant differences in the expression levels of IRE1 and PERK proteins in control(CT),sham(SH)-,RF-and GR-exposed cells(P<0.05).The phosphorylated protein levels of p-IRE1 and p-PERK were significantly increased in cells exposed to RF and GR(P<0.05).The expression levels of GRP78 and CHOP were significantly increased in RF-and GR-exposed cells compared to CT and SH-exposed cells(P<0.05).Cells treated with 1μg/ml TM for 24 h showed significantly increased expression levels of GRP78 and CHOP proteins compared to controls(P<0.05).In the presence of 2 mmol/L PBA,TM-induced increased levels of GRP78 and CHOP proteins were reduced(P<0.05).Conclusions:The exposure of non-ionizing 900 MHz RF was able to cause stress in HT22 mouse neuronal cells and activated UPR in ER.Since UPR plays an important role in both cell survival(when UPR is mitigated)and apoptosis/death(under unresolvable stress conditions),further studies are required to determine the fate of the cells exposed to RF.
文摘本研究探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对自然衰老大鼠内质网应激相关蛋白的调节作用以及对神经细胞凋亡的干预作用。雄性SD大鼠随机分为6月龄组、24月龄组、SPJ低剂量组(10mg/kg)和高剂量组(30mg/kg)。Westernblot法检测大鼠皮层和海马内质网应激相关蛋白(GRP78、p-PERK、p-EIF2α、ATF4、ATF6、p-IRE1、XBP1)以及凋亡相关蛋白(p-JNK、CHOP、Bcl-2、Bax)表达变化。结果显示,与6月龄组相比,24月龄组皮层/海马组织中GRP78、p-PERK、p-EIF2α、ATF4、ATF6、p-IRE1、XBP1和Bcl-2/Bax蛋白表达分别下降了80%/59%、82%/78%、79%/77%、94%/74%、72%/66%、78%/81%、82%/71%和90%/92%,p-JNK/JNK、CHOP蛋白表达分别上升了1/1.72、3.33/1.33倍。给予不同剂量竹节参总皂苷干预后,均能明显上调内质网应激相关蛋白和Bcl-2/Bax的表达水平,下调促凋亡蛋白表达水平。研究结果表明,竹节参总皂苷能通过上调内质网应激相关蛋白以及抗凋亡蛋白的表达水平,下调促凋亡蛋白的表达,从而维持衰老大鼠脑组织的内质网稳态,减少神经细胞凋亡。
文摘目的探讨CAAT增强子结合蛋白同源蛋白(CHOP)、海马葡萄糖调节蛋白78(GRP78)和caspase-12在蛛网膜下腔出血(SAH)后的表达及其在早期脑损伤(EBI)中的作用。方法选取健康雄性SD大鼠78只,采用随机数字表法将其分为空白对照组(6只)、假手术组(36只)与SAH组(36只)。将假手术组和SAH组分别于1、6、12、24、48、72h,采用简单随机抽样法选取6只大鼠,神经功能缺陷评分后处死。比较各组不同时间点光镜及电镜下大鼠海马神经元形态改变、CHOP、GRP78、caspase-12相对表达水平、海马神经元凋亡细胞水平、脑水肿严重程度及神经功能缺陷评分情况。结果通过Western blot法检测CHOP、GRP78和caspase-12的表达,SAH组与空白对照组、假手术组在12、24、48、72h比较,差异均有统计学意义(P<0.05),且在SAH后24h达到峰值。TUNEL法检测大鼠海马区神经元凋亡显示,SAH组与对照组、假手术组在6、12、24、48、72h比较,差异均有统计学意义(P<0.05),在电镜下,SAH后24h节点可见明显的神经元凋亡。SAH组12、24、48、72h的神经功能缺陷评分及脑水肿程度明显重于空白对照组及假手术组,在24h达高峰,差异均有统计学意义(P<0.05)。SAH组大鼠的CHOP、GRP78和caspase-12 Western blot法检测结果与脑水肿严重程度及神经元凋亡结果之间呈极显著正相关,与神经功能缺陷评分呈极显著负相关(P<0.05)。结论 SAH后72h内确实造成了EBI,同时发生了内质网应激,内质网应激与EBI严重程度之间均呈正相关,揭示内质网应激在SAH后EBI的病理过程中发挥了重要作用。
文摘【目的】探讨四逆汤对阿霉素诱导的慢性心力衰竭(CHF)大鼠内质网应激的影响。【方法】从90只大鼠中随机选取15只作为正常组,其余大鼠腹腔注射阿霉素构建CHF模型,同时,正常组给予腹腔注射生理盐水。造模结束后,正常组大鼠全部存活,造模大鼠存活60只。再将60只CHF大鼠随机分为5组,即模型组,四逆汤低、中、高剂量组,曲美他嗪组,每组12只。四逆汤低、中、高剂量组分别给予1.4、4.2、12.6(生药)g·kg^(-1)·d^(-1)灌胃,曲美他嗪组给予曲美他嗪10 mg·kg^(-1)·d^(-1)灌胃,正常组和模型组给予相同体积的生理盐水灌胃。连续给药4周。采用小动物超声测量左心室舒张末期直径(LVEDD)、左心室射血分数(LVEF)和左心室缩短分数(LVFS)变化;采用酶联免疫吸附分析(ELISA)检测血浆脑钠肽(BNP)和血管紧张素Ⅱ(AngⅡ)水平;采用实时荧光定量聚合酶链反应(PCR)技术检测心肌组织葡萄糖调节蛋白78(GRP78)、RNA依赖的蛋白激酶样内质网激酶(PERK)和活化转录因子4(ATF4)mRNA表达。【结果】与正常组比较,模型组大鼠LVEDD显著增大,LVEF、LVFS均显著降低,血浆BNP和AngⅡ含量升高,心肌组织GRP78、PERK及ATF4 m RNA表达量升高(P <0.01);与模型组比较,四逆汤低、中、高剂量组及曲美他嗪组LVEDD降低,LVEF、LVFS均显著增加,血浆BNP和AngⅡ含量减少,心肌组织GRP78、PERK及ATF4 m RNA表达量降低,且呈剂量依赖性,其中,四逆汤高剂量组及曲美他嗪组差异均有统计学意义(P <0.05或P <0.01)。【结论】四逆汤可改善慢性心力衰竭大鼠模型心脏功能,其机制可能与抑制PERK/ATF4信号通路从而减轻内质网应激有关。
基金supported by grants from the National Basic ResearchProgram(973)(No.2012CB517503,No.2011CB503903,and No.2012CB945003)National Natural Science Foundation of China(No.81070120)to Qi Chen+1 种基金National Natural Science Foundation ofChina(No.81000118)University Natural Science Foundation ofJiangsu(No.10KJB310005)to Jingjing Ben
文摘Class A scavenger receptor （SR-A） plays an important role in macrophage adhesion. However, the underlying mechanism remains unclear. We previously found that 78 kDa glucose-regulated protein （GRP78） inhibited SR- A-mediated ligand internalization into macrophage by binding to SR-A. The aim of the study was to investigate whether GRP78 could regulate SR-A-mediated cell adhesion. We demonstrated that GRP78 bound directly to SR-A by fluorescence resonance energy transfer （FRET） assay. Overexpression of GRP78 inhibited macrophage adhesion via SR-A. These results suggest that GRP78 may act as an inhibitor of macrophage adhesion via SR-A.
