目的探讨Salubrinal对大鼠脑缺血再灌注模型LC3-ⅡmRNA及LC3-Ⅱ/LC3-ⅠmRNA表达的影响。方法用线栓法建立大鼠大脑中动脉脑缺血(MCAO)缺血再灌注模型。大鼠随机分为假手术组、模型组、Salubrinal组(Sal组),各组又分为再灌6、12、24、72 ...目的探讨Salubrinal对大鼠脑缺血再灌注模型LC3-ⅡmRNA及LC3-Ⅱ/LC3-ⅠmRNA表达的影响。方法用线栓法建立大鼠大脑中动脉脑缺血(MCAO)缺血再灌注模型。大鼠随机分为假手术组、模型组、Salubrinal组(Sal组),各组又分为再灌6、12、24、72 h 4个亚组。各组于再灌相应时间点行神经功能缺损评分,并断头取脑行HE染色及Real time-PCR检测。结果神经功能缺损:与假手术组比较,模型组、Sal组各时间点均有神经功能缺损(P<0.01);与模型组比较,再灌24、72 h Sal组神经功能缺损评分降低(P<0.05)。HE染色:模型组神经元减少、缺失,细胞肿胀,部分破裂,细胞核固缩、偏移、深染,胶质细胞增生,经Salubrinal干预后,再灌各时间点神经元增多,胞膜较完整,核固缩现象较轻,水肿轻微。Real time-PCR检测:与假手术组比较,模型组、Sal组各指标于再灌各时间点均增高(P<0.01)。与模型组比,Sal组LC3-ⅡmRNA表达于再灌12、24、72 h下降明显(P<0.05);Sal组LC3-Ⅱ/LC3-ⅠmRNA于再灌后各时间点下降均明显(P<0.01)。结论 Salubrinal通过下调LC3-ⅡmRNA及LC3-Ⅱ/LC3-ⅠmRNA比值,对大鼠脑缺血再灌注损伤起保护作用。展开更多
Objectives This study examined the protective effect of salubrinal and the mechanism underlying this protection against tunicamycin （TM）- and hypoxia-induced apoptosis in rat cardiomyocytes. Methods Neonatal rat car...Objectives This study examined the protective effect of salubrinal and the mechanism underlying this protection against tunicamycin （TM）- and hypoxia-induced apoptosis in rat cardiomyocytes. Methods Neonatal rat cardiomyocytes were cultured from the ventricles of l-day-old Wistar rats. Cells were exposed to different concentrations of salubrinal （10, 20, and 40 gmol/L） for 30 min followed by TM treatment or hypoxia for 36 h. Apoptosis was measured by a multiparameter HCS （high content screening） apoptosis assay, TUNEL assay and flow cytometry. The phosphorylation of eukaryotic translation initiation factor 2 subunit alpha （eIF2c0 and the expression of cleaved caspase-12 were determined by Western blotting. C/EBP homologous protein （CHOP） was detected by immunocytochemistry. Results HCS, TUNEL assays and flow cytometry showed that salubrinal protected cardiomyocytes against apoptosis induced by TM or hypoxia. Western blotting showed that salubrinal protected cardiomyocytes against apoptosis by inducing eIF2ct phosphorylation and down-regulating the expression of the endoplasmic reticulum stress-mediated apoptotic proteins, CHOP and cleaved caspase-12. Conclusions Our study suggests that salubrinal protects rat cardiomyocytes against TM- or hypoxia-associated apoptosis via a mechanism involving the inhibition of ER stress-mediated apoptosis.展开更多
Most common cerebrovascular accidents （CVAs） result from the occlusion of a blood vessel and are called ischemic stroke. The interruption of the blood flow reduces the oxygen and glucose levels in the neural pareuch...Most common cerebrovascular accidents （CVAs） result from the occlusion of a blood vessel and are called ischemic stroke. The interruption of the blood flow reduces the oxygen and glucose levels in the neural pareuchyma, which in turn decreases the energy of the cells and compromises their homeostasis. This condition has to be reverted for cell survival. One of the homeostatic processes impaired by cerebral ischemia is protein folding in the endoplasmic reticulum （ER） lumen where unfolded proteins are accumulated （Degracia and Monti, 2004）. This situation, known as ER stress, elicits the so called unfolded protein response （UPR）,展开更多
Spinal cord injury(SCI)remains an intractable clinical challenge of neurosurgery,it can be divided into two stages:uncontrollable primary injury induced by mechanical damage and controllable secondary injury regulated...Spinal cord injury(SCI)remains an intractable clinical challenge of neurosurgery,it can be divided into two stages:uncontrollable primary injury induced by mechanical damage and controllable secondary injury regulated by continuous cell death.The apoptosis was the one of most important events in secondary injury,previous studies revealed that excessive endoplasmic reticulum(ER)stress breaks down the homeostasis and triggers apoptosis in the spinal cord.To deter or alleviate the secondary jury,we screen one of fat-soluble compounds,salubrinal,which was an inhibitor of eIF2αdephosphorylation can repair SCI by inhibiting ER stress in mice after SCI.Administration of salubrinal effectively represses apoptosis,protects neuronal cell,and promotes the restoration of locomotor function in mice SCI models.Furthermore,the level of phosphorylated eIF2αwas raised in the presence of salubrinal,but the protein expression of ATF4 and CHOP was downregulated.Unexpectedly,transcriptional expression of CHOPregulated pro-apoptotic genes was decreased.These data suggest salubrinal suppress ER stress by targeting eIF2α/ATF4 pathways and reduces cell death after SCI.It is suggested that the mitigation of secondary lesion by inhibiting ER stress induced apoptosis in the early phase of SCI is promising treatment strategy.展开更多
Background： Previous studies have indicated that endoplasmic reticulum stress participates in and mediates liver injury and apoptosis in brain-dead （BD） rats. In this study, we observed the effect ofsalubrinal （Sa...Background： Previous studies have indicated that endoplasmic reticulum stress participates in and mediates liver injury and apoptosis in brain-dead （BD） rats. In this study, we observed the effect ofsalubrinal （Sal, Sigma, USA） on liver cells in BD rats and explored its relevant mechanisms. Methods： Thirty Sprague-Dawley rats were equally randomized into three groups： BD group, Sal group, and DMSO group. The BD models were established by increasing intracranial pressure in a modified, slow, and intermittent way. In the drug groups, Sal was administered I h before the induction of BD. After modeling was completed, the blood and liver samples were harvested. CHOP and Caspase-12 mRNA expression was detected using quantitative polymerase chain reaction. PKR-like ER kinase （PERK）, P-eukaryotic translation initiation factor 2a （elF2a）, elF2a, CHOP and caspase-12 expression was detected using western blotting （WB）. CHOP and caspase-12 distribution and expression in liver tissues were determined using immunohistochemistry （IHC）. Alanine aminotransferase and aspartate aminotransferase level were detected using an automatic biochemical analyzer. Hepatic cell apoptosis was detected using TUNEL. The results were analyzed using Quantity-one v4.62 software （Bio-Rad, USA）. Results： CHOP and caspase-12 expression and PERK, elF2a, and P-elF2a protein expression showed no significant difference between BD group and DMSO group. Compared with BD group, Sal group had a significantly higher P-elF2C level and a lower P-PERK level 2 h and 6 h alter BD （P 〈 0.05）. However, eIF2a expression showed no significant difference （P 〉 0.05）. After the Sal treatment, CHOP and caspase- 12 mRNA expression significantly decreased 4 hatter BD （P 〈 0.05）. WB and IHC indicated that CHOP and caspase- 12 expression also significantly decreased after Sal treatment. Sal was associated with improved liver function and decreased hepatic cell apoptosis. Conclusions： Sal can significantly reduce apoptosis in hepatic cells of BD rats. This protective effect may be achieved via the PERK-elF2a signaling pathway.展开更多
文摘目的探讨Salubrinal对大鼠脑缺血再灌注模型LC3-ⅡmRNA及LC3-Ⅱ/LC3-ⅠmRNA表达的影响。方法用线栓法建立大鼠大脑中动脉脑缺血(MCAO)缺血再灌注模型。大鼠随机分为假手术组、模型组、Salubrinal组(Sal组),各组又分为再灌6、12、24、72 h 4个亚组。各组于再灌相应时间点行神经功能缺损评分,并断头取脑行HE染色及Real time-PCR检测。结果神经功能缺损:与假手术组比较,模型组、Sal组各时间点均有神经功能缺损(P<0.01);与模型组比较,再灌24、72 h Sal组神经功能缺损评分降低(P<0.05)。HE染色:模型组神经元减少、缺失,细胞肿胀,部分破裂,细胞核固缩、偏移、深染,胶质细胞增生,经Salubrinal干预后,再灌各时间点神经元增多,胞膜较完整,核固缩现象较轻,水肿轻微。Real time-PCR检测:与假手术组比较,模型组、Sal组各指标于再灌各时间点均增高(P<0.01)。与模型组比,Sal组LC3-ⅡmRNA表达于再灌12、24、72 h下降明显(P<0.05);Sal组LC3-Ⅱ/LC3-ⅠmRNA于再灌后各时间点下降均明显(P<0.01)。结论 Salubrinal通过下调LC3-ⅡmRNA及LC3-Ⅱ/LC3-ⅠmRNA比值,对大鼠脑缺血再灌注损伤起保护作用。
基金This study was supported by the Ministry Science Foundation of the Chinese People's Liberation Army during the 12th Five-Year Plan Period
文摘Objectives This study examined the protective effect of salubrinal and the mechanism underlying this protection against tunicamycin （TM）- and hypoxia-induced apoptosis in rat cardiomyocytes. Methods Neonatal rat cardiomyocytes were cultured from the ventricles of l-day-old Wistar rats. Cells were exposed to different concentrations of salubrinal （10, 20, and 40 gmol/L） for 30 min followed by TM treatment or hypoxia for 36 h. Apoptosis was measured by a multiparameter HCS （high content screening） apoptosis assay, TUNEL assay and flow cytometry. The phosphorylation of eukaryotic translation initiation factor 2 subunit alpha （eIF2c0 and the expression of cleaved caspase-12 were determined by Western blotting. C/EBP homologous protein （CHOP） was detected by immunocytochemistry. Results HCS, TUNEL assays and flow cytometry showed that salubrinal protected cardiomyocytes against apoptosis induced by TM or hypoxia. Western blotting showed that salubrinal protected cardiomyocytes against apoptosis by inducing eIF2ct phosphorylation and down-regulating the expression of the endoplasmic reticulum stress-mediated apoptotic proteins, CHOP and cleaved caspase-12. Conclusions Our study suggests that salubrinal protects rat cardiomyocytes against TM- or hypoxia-associated apoptosis via a mechanism involving the inhibition of ER stress-mediated apoptosis.
文摘Most common cerebrovascular accidents （CVAs） result from the occlusion of a blood vessel and are called ischemic stroke. The interruption of the blood flow reduces the oxygen and glucose levels in the neural pareuchyma, which in turn decreases the energy of the cells and compromises their homeostasis. This condition has to be reverted for cell survival. One of the homeostatic processes impaired by cerebral ischemia is protein folding in the endoplasmic reticulum （ER） lumen where unfolded proteins are accumulated （Degracia and Monti, 2004）. This situation, known as ER stress, elicits the so called unfolded protein response （UPR）,
基金This study was supported by Anhui Natural Science Foundation(1908085MC83)the National Natural Science Foundation of China(31970413)Wuhu Science and Technology Bureau Project(2019cg19).
文摘Spinal cord injury(SCI)remains an intractable clinical challenge of neurosurgery,it can be divided into two stages:uncontrollable primary injury induced by mechanical damage and controllable secondary injury regulated by continuous cell death.The apoptosis was the one of most important events in secondary injury,previous studies revealed that excessive endoplasmic reticulum(ER)stress breaks down the homeostasis and triggers apoptosis in the spinal cord.To deter or alleviate the secondary jury,we screen one of fat-soluble compounds,salubrinal,which was an inhibitor of eIF2αdephosphorylation can repair SCI by inhibiting ER stress in mice after SCI.Administration of salubrinal effectively represses apoptosis,protects neuronal cell,and promotes the restoration of locomotor function in mice SCI models.Furthermore,the level of phosphorylated eIF2αwas raised in the presence of salubrinal,but the protein expression of ATF4 and CHOP was downregulated.Unexpectedly,transcriptional expression of CHOPregulated pro-apoptotic genes was decreased.These data suggest salubrinal suppress ER stress by targeting eIF2α/ATF4 pathways and reduces cell death after SCI.It is suggested that the mitigation of secondary lesion by inhibiting ER stress induced apoptosis in the early phase of SCI is promising treatment strategy.
基金This study was supported by a grant of National Natural Science Foundation of China （No. 81171849）.
文摘Background： Previous studies have indicated that endoplasmic reticulum stress participates in and mediates liver injury and apoptosis in brain-dead （BD） rats. In this study, we observed the effect ofsalubrinal （Sal, Sigma, USA） on liver cells in BD rats and explored its relevant mechanisms. Methods： Thirty Sprague-Dawley rats were equally randomized into three groups： BD group, Sal group, and DMSO group. The BD models were established by increasing intracranial pressure in a modified, slow, and intermittent way. In the drug groups, Sal was administered I h before the induction of BD. After modeling was completed, the blood and liver samples were harvested. CHOP and Caspase-12 mRNA expression was detected using quantitative polymerase chain reaction. PKR-like ER kinase （PERK）, P-eukaryotic translation initiation factor 2a （elF2a）, elF2a, CHOP and caspase-12 expression was detected using western blotting （WB）. CHOP and caspase-12 distribution and expression in liver tissues were determined using immunohistochemistry （IHC）. Alanine aminotransferase and aspartate aminotransferase level were detected using an automatic biochemical analyzer. Hepatic cell apoptosis was detected using TUNEL. The results were analyzed using Quantity-one v4.62 software （Bio-Rad, USA）. Results： CHOP and caspase-12 expression and PERK, elF2a, and P-elF2a protein expression showed no significant difference between BD group and DMSO group. Compared with BD group, Sal group had a significantly higher P-elF2C level and a lower P-PERK level 2 h and 6 h alter BD （P 〈 0.05）. However, eIF2a expression showed no significant difference （P 〉 0.05）. After the Sal treatment, CHOP and caspase- 12 mRNA expression significantly decreased 4 hatter BD （P 〈 0.05）. WB and IHC indicated that CHOP and caspase- 12 expression also significantly decreased after Sal treatment. Sal was associated with improved liver function and decreased hepatic cell apoptosis. Conclusions： Sal can significantly reduce apoptosis in hepatic cells of BD rats. This protective effect may be achieved via the PERK-elF2a signaling pathway.
