The endoplasmic reticulum is the central organelle within a eukaryotic cell where newly synthesized proteins are processed and properly folded. An excess of unfolded or mis-folded proteins induces ER stress signalling...The endoplasmic reticulum is the central organelle within a eukaryotic cell where newly synthesized proteins are processed and properly folded. An excess of unfolded or mis-folded proteins induces ER stress signalling pathways. Usually this means a pro-survival strategy for the cell, whereas under extended stress conditions the ER stress signalling pathways have a pro-apoptotic function. CK2 plays a key role in the regulation of the pro-survival as well as the proapoptotic ER stress signalling by directly modulating the activities of members of the ER stress signalling pathways by phosphorylation, regulating the expression of the key factors of the signalling pathways or binding to regulator proteins. The present review will summarize the state of the art in this new emerging field.展开更多
目的:探讨蛋白激酶RNA样ER激酶(protein kinase RNA-like ER kinase,PERK)信号通路介导的线粒体未折叠蛋白反应(mitochondrial unfolded protein response,mtUPR)在缺氧缺血性脑损伤(hypoxic-ischemic brain injury,HIBI)中的作用。方法...目的:探讨蛋白激酶RNA样ER激酶(protein kinase RNA-like ER kinase,PERK)信号通路介导的线粒体未折叠蛋白反应(mitochondrial unfolded protein response,mtUPR)在缺氧缺血性脑损伤(hypoxic-ischemic brain injury,HIBI)中的作用。方法:将大鼠随机分为假手术(Sham)组和5个HIBI亚组(HIBI后3、6、12、24、48 h)。用于蛋白质印迹检测PERK、转录激活因子4(activating transcription factor 4,ATF4)、热休克蛋白60(heat shock protein 60,HSP60)蛋白的时程表达。将大鼠随机分为Sham组、HIBI组、HIBI+PERK组和HIBI+载体(Vector)组,每组15只。HIBI+PERK组和HIBI+Vector组大鼠在HIBI手术前1 h,将基于腺病毒相关病毒(adeno-associated virus,AAV)的PERK过表达质粒或AAV载体注射到脑室内,用于特异性表达PERK。在HIBI后24 h进行FJC染色分析神经元变性和DHE染色、酶联免疫吸附试验分析氧化应激。将大鼠随机分为Sham组、HIBI组、HIBI+PERK激动剂(CCT020312)组,每组12只。在HIBI手术前1 h,向HIBI+CCT020312组大鼠脑室内注射CCT020312。在HIBI后3周进行开阔场地测试和莫里斯水迷宫测试。结果:与Sham组相比,PERK、ATF4、HSP60在HIBI后3 h开始明显升高,在12 h达到高峰,然后逐渐下降,直到48 h(F=60.23、56.72、74.31,均P<0.001)。与HIBI组相比,HIBI+PERK组神经元变性的数量(100.2±3.1 vs. 582.4±15.7,P<0.001)、活性氧(reactive oxygen species,ROS)(42.4±2.9 vs. 17.7±2.1,P<0.01)、丙二醛(Malondialdehyde,MDA)(0.81±0.06 vs. 0.54±0.04,P<0.001)水平显著降低,和谷胱甘肽过氧化物酶(glutathione peroxidase,GSHPx)(112.4±3.6 vs. 177.5±6.6,P<0.05)、超氧化物歧化酶(superoxide Dismutase,SOD)活性(46.3±1.9 vs. 64.2±2.3,P<0.05)活性明显增加。与Sham组相比,HIBI组大鼠海马组织中PERK(1.00±0.03 vs. 1.66±0.08,P<0.01)、ATF4(1.00±0.04 vs.1.53±0.06,P<0.05)、动力蛋白相关蛋白1(dynamin-related protein 1,Drp1)(1.00±0.02 vs. 1.98±0.07,P<0.01)、HSP60(1.00±0.03 vs. 1.37±0.04,P<0.05)蛋白表达均明显增加(P<0.05)。与HIBI组相比,HIBI+PERK组大鼠海马组织中PERK(1.66±0.08vs. 2.95±0.17,P<0.01)、ATF4(1.53±0.06 vs. 3.42±0.22,P<0.01)、HSP60(1.37±0.04 vs. 2.03±0.09,P<0.05)蛋白表达均明显增加(F=46.72、30.63、20.64,P<0.001),和Drp1(1.98±0.07 vs. 1.04±0.05,P<0.05)蛋白表达明显降低(F=35.72,P<0.001)。HIBI+CCT020312组的平均逃避潜伏期和平台穿越次数均较HIBI组明显增加(F=246.84、113.62,P<0.001)。结论:PERK减轻HIBI模型诱导的氧化应激和神经元凋亡,其机制可能涉及PERK/ATF4信号通路对mtUPR的调节。通过CCT020312给药具有神经保护作用。展开更多
Heat shock proteins (HSPs) serve to correct proteins’ conformation, send the damaged proteins for degradation (quality control function). Heat shock factors (HSFs) are their transcription factors. The protein complex...Heat shock proteins (HSPs) serve to correct proteins’ conformation, send the damaged proteins for degradation (quality control function). Heat shock factors (HSFs) are their transcription factors. The protein complexes mTOR1 and 2 (with the same core mTOR), the phosphoinositide-dependent protein kinase-1 (PDK1), the seine/threonine-specific protein kinase (Akt), HSF1, plus their associated proteins form a network participating in protein synthesis, bio-energy generation, signaling for apoptosis with the help of HSPs. A cancer cell synthesizes proteins at fast rate and needs more HSPs to work on quality control. Shutting down this network would lead to cell death. Thus inhibitors of mTOR (mTORI) and inhibitors of HSPs (HSPI) could drive cancer cell to apoptosis—a “passive approach”. On the other hand, HSPs form complexes with polypeptides characteristic of the cancer cells;on excretion from the cell, they becomes antigens for the immunity cells, eventually leading to maturation of the cytotoxic T cells, forming the basic principle of preparing cancer-specific, person-specific vaccine. Recent finding shows that HSP70 can penetrate cancer cell and expel its analog to extracellular region, giving the hope to prepare a non-person-specific vaccine covering a variety of cancers. Activation of anti-cancer immunity is the “active approach”. On the other hand, mild hyperthermia, with increase of intracellular HSPs, has been found to activate the immunity response, and demonstrate anti-cancer effects. There are certain “mysteries” behind the mechanisms of the active and passive approaches. We analyze the mechanisms involved and provide explanations to some mysteries. We also suggest future research to improve our understanding of these two approaches, in which HSPs play many roles.展开更多
[Objectives] To study the therapeutic effect and mechanism of New Maixian Powder on ulcerative colitis( UC) rats through observing its regulatory effect on the protein kinase R-like endoplasmic reticulum kinase( PERK)...[Objectives] To study the therapeutic effect and mechanism of New Maixian Powder on ulcerative colitis( UC) rats through observing its regulatory effect on the protein kinase R-like endoplasmic reticulum kinase( PERK)/eukaryotic translation initiation factor-2α( e IF-2α)/nuclear transcription factor-kappa B( NF-κB) signaling pathway. [Methods]First,60 SD rats were randomly divided into normal group,model group,mesalazine group,and New Maixian Powder low,medium and high dose groups,10 rats each group. Then,dextran sulfate sodium( DSS) was used to induce UC rats. The mesalazine group was given 0. 42 g/( kg·d) of mesalazine sustained-release granule suspension,New Maixian Powder low,medium and high dose groups were given 1. 5,3,and 6 g/( kg·d) of New Maixian Powder suspension,respectively,and other groups were given an equal volume of physiological saline,continuous intragastric administration for 14 d. Next,the disease activity index( DAI) of UC rats was evaluated; the expression of NF-κB in serum was measured by enzyme-linked immunosorbent assay( ELISA); the expression of PERK and e IF-2α protein and m RNA in colon tissue was detected by Western blot and real-time quantitative polymerase chain reaction( RT q-PCR). [Results] Compared with the normal group,the DAI score and serum NF-κB level in the model group were significantly higher( P < 0. 05),and PERK and e IF-2α protein and m RNA levels in the colon tissue were increased( P < 0. 05); compared with the model group,the DAI score decreased and serum NF-κB level declined in the New Maixian Powder group,and the expression of PERK and e IF-2α protein and m RNA in New Maixian Powder medium dose and high dose groups declined( P < 0. 05). [Conclusions]New Maixian Powder has good therapeutic effect on UC rats,and its mechanism may be connected with the inhibition of the activation of PERK/e IF-2α/NF-κB signaling pathway.展开更多
Both environmental and genetic factors contribute to the development of diabetes mellitus and although monogenic disorders are rare,they offer unique insights into the fundamental biology underlying the disease.Mutati...Both environmental and genetic factors contribute to the development of diabetes mellitus and although monogenic disorders are rare,they offer unique insights into the fundamental biology underlying the disease.Mutations of the insulin gene or genes involved in the response to protein misfolding cause early onset diabetes.These have revealed an important role for endoplasmic reticulum stress in β-cell survival.This form of cellular stress occurs when secretory proteins fail to fold efficiently.Of all the professional secretory cells we possess,β-cells are the most sensitive to endoplasmic reticulum stress because of the large fluctuations in protein synthesis they face daily.Studies of endoplasmic reticulum stress signaling therefore offer the potential to identify new drug targets to treat diabetes.展开更多
文摘The endoplasmic reticulum is the central organelle within a eukaryotic cell where newly synthesized proteins are processed and properly folded. An excess of unfolded or mis-folded proteins induces ER stress signalling pathways. Usually this means a pro-survival strategy for the cell, whereas under extended stress conditions the ER stress signalling pathways have a pro-apoptotic function. CK2 plays a key role in the regulation of the pro-survival as well as the proapoptotic ER stress signalling by directly modulating the activities of members of the ER stress signalling pathways by phosphorylation, regulating the expression of the key factors of the signalling pathways or binding to regulator proteins. The present review will summarize the state of the art in this new emerging field.
文摘目的:探讨蛋白激酶RNA样ER激酶(protein kinase RNA-like ER kinase,PERK)信号通路介导的线粒体未折叠蛋白反应(mitochondrial unfolded protein response,mtUPR)在缺氧缺血性脑损伤(hypoxic-ischemic brain injury,HIBI)中的作用。方法:将大鼠随机分为假手术(Sham)组和5个HIBI亚组(HIBI后3、6、12、24、48 h)。用于蛋白质印迹检测PERK、转录激活因子4(activating transcription factor 4,ATF4)、热休克蛋白60(heat shock protein 60,HSP60)蛋白的时程表达。将大鼠随机分为Sham组、HIBI组、HIBI+PERK组和HIBI+载体(Vector)组,每组15只。HIBI+PERK组和HIBI+Vector组大鼠在HIBI手术前1 h,将基于腺病毒相关病毒(adeno-associated virus,AAV)的PERK过表达质粒或AAV载体注射到脑室内,用于特异性表达PERK。在HIBI后24 h进行FJC染色分析神经元变性和DHE染色、酶联免疫吸附试验分析氧化应激。将大鼠随机分为Sham组、HIBI组、HIBI+PERK激动剂(CCT020312)组,每组12只。在HIBI手术前1 h,向HIBI+CCT020312组大鼠脑室内注射CCT020312。在HIBI后3周进行开阔场地测试和莫里斯水迷宫测试。结果:与Sham组相比,PERK、ATF4、HSP60在HIBI后3 h开始明显升高,在12 h达到高峰,然后逐渐下降,直到48 h(F=60.23、56.72、74.31,均P<0.001)。与HIBI组相比,HIBI+PERK组神经元变性的数量(100.2±3.1 vs. 582.4±15.7,P<0.001)、活性氧(reactive oxygen species,ROS)(42.4±2.9 vs. 17.7±2.1,P<0.01)、丙二醛(Malondialdehyde,MDA)(0.81±0.06 vs. 0.54±0.04,P<0.001)水平显著降低,和谷胱甘肽过氧化物酶(glutathione peroxidase,GSHPx)(112.4±3.6 vs. 177.5±6.6,P<0.05)、超氧化物歧化酶(superoxide Dismutase,SOD)活性(46.3±1.9 vs. 64.2±2.3,P<0.05)活性明显增加。与Sham组相比,HIBI组大鼠海马组织中PERK(1.00±0.03 vs. 1.66±0.08,P<0.01)、ATF4(1.00±0.04 vs.1.53±0.06,P<0.05)、动力蛋白相关蛋白1(dynamin-related protein 1,Drp1)(1.00±0.02 vs. 1.98±0.07,P<0.01)、HSP60(1.00±0.03 vs. 1.37±0.04,P<0.05)蛋白表达均明显增加(P<0.05)。与HIBI组相比,HIBI+PERK组大鼠海马组织中PERK(1.66±0.08vs. 2.95±0.17,P<0.01)、ATF4(1.53±0.06 vs. 3.42±0.22,P<0.01)、HSP60(1.37±0.04 vs. 2.03±0.09,P<0.05)蛋白表达均明显增加(F=46.72、30.63、20.64,P<0.001),和Drp1(1.98±0.07 vs. 1.04±0.05,P<0.05)蛋白表达明显降低(F=35.72,P<0.001)。HIBI+CCT020312组的平均逃避潜伏期和平台穿越次数均较HIBI组明显增加(F=246.84、113.62,P<0.001)。结论:PERK减轻HIBI模型诱导的氧化应激和神经元凋亡,其机制可能涉及PERK/ATF4信号通路对mtUPR的调节。通过CCT020312给药具有神经保护作用。
文摘Heat shock proteins (HSPs) serve to correct proteins’ conformation, send the damaged proteins for degradation (quality control function). Heat shock factors (HSFs) are their transcription factors. The protein complexes mTOR1 and 2 (with the same core mTOR), the phosphoinositide-dependent protein kinase-1 (PDK1), the seine/threonine-specific protein kinase (Akt), HSF1, plus their associated proteins form a network participating in protein synthesis, bio-energy generation, signaling for apoptosis with the help of HSPs. A cancer cell synthesizes proteins at fast rate and needs more HSPs to work on quality control. Shutting down this network would lead to cell death. Thus inhibitors of mTOR (mTORI) and inhibitors of HSPs (HSPI) could drive cancer cell to apoptosis—a “passive approach”. On the other hand, HSPs form complexes with polypeptides characteristic of the cancer cells;on excretion from the cell, they becomes antigens for the immunity cells, eventually leading to maturation of the cytotoxic T cells, forming the basic principle of preparing cancer-specific, person-specific vaccine. Recent finding shows that HSP70 can penetrate cancer cell and expel its analog to extracellular region, giving the hope to prepare a non-person-specific vaccine covering a variety of cancers. Activation of anti-cancer immunity is the “active approach”. On the other hand, mild hyperthermia, with increase of intracellular HSPs, has been found to activate the immunity response, and demonstrate anti-cancer effects. There are certain “mysteries” behind the mechanisms of the active and passive approaches. We analyze the mechanisms involved and provide explanations to some mysteries. We also suggest future research to improve our understanding of these two approaches, in which HSPs play many roles.
基金Supported by Science and Technology Program of Zhejiang Province(2016C33085)
文摘[Objectives] To study the therapeutic effect and mechanism of New Maixian Powder on ulcerative colitis( UC) rats through observing its regulatory effect on the protein kinase R-like endoplasmic reticulum kinase( PERK)/eukaryotic translation initiation factor-2α( e IF-2α)/nuclear transcription factor-kappa B( NF-κB) signaling pathway. [Methods]First,60 SD rats were randomly divided into normal group,model group,mesalazine group,and New Maixian Powder low,medium and high dose groups,10 rats each group. Then,dextran sulfate sodium( DSS) was used to induce UC rats. The mesalazine group was given 0. 42 g/( kg·d) of mesalazine sustained-release granule suspension,New Maixian Powder low,medium and high dose groups were given 1. 5,3,and 6 g/( kg·d) of New Maixian Powder suspension,respectively,and other groups were given an equal volume of physiological saline,continuous intragastric administration for 14 d. Next,the disease activity index( DAI) of UC rats was evaluated; the expression of NF-κB in serum was measured by enzyme-linked immunosorbent assay( ELISA); the expression of PERK and e IF-2α protein and m RNA in colon tissue was detected by Western blot and real-time quantitative polymerase chain reaction( RT q-PCR). [Results] Compared with the normal group,the DAI score and serum NF-κB level in the model group were significantly higher( P < 0. 05),and PERK and e IF-2α protein and m RNA levels in the colon tissue were increased( P < 0. 05); compared with the model group,the DAI score decreased and serum NF-κB level declined in the New Maixian Powder group,and the expression of PERK and e IF-2α protein and m RNA in New Maixian Powder medium dose and high dose groups declined( P < 0. 05). [Conclusions]New Maixian Powder has good therapeutic effect on UC rats,and its mechanism may be connected with the inhibition of the activation of PERK/e IF-2α/NF-κB signaling pathway.
基金Supported by a PhD studentship form Diabetes UK (for Thomas SE)
文摘Both environmental and genetic factors contribute to the development of diabetes mellitus and although monogenic disorders are rare,they offer unique insights into the fundamental biology underlying the disease.Mutations of the insulin gene or genes involved in the response to protein misfolding cause early onset diabetes.These have revealed an important role for endoplasmic reticulum stress in β-cell survival.This form of cellular stress occurs when secretory proteins fail to fold efficiently.Of all the professional secretory cells we possess,β-cells are the most sensitive to endoplasmic reticulum stress because of the large fluctuations in protein synthesis they face daily.Studies of endoplasmic reticulum stress signaling therefore offer the potential to identify new drug targets to treat diabetes.
