Objective:Myocardial ischemia-reperfusion injury(MIRI)is one of the leading causes of death from cardiovascular disease in humans,especially in individuals exposed to cold environments.Long non-coding RNAs(lncRNAs)reg...Objective:Myocardial ischemia-reperfusion injury(MIRI)is one of the leading causes of death from cardiovascular disease in humans,especially in individuals exposed to cold environments.Long non-coding RNAs(lncRNAs)regulate MIRI through multiple mechanisms.This study explored the regulatory effect of lncRNA-AK138945 on myocardial ischemia-reperfusion injury and its mechanism.Methods:In vivo,8-to 12-weeks-old C57BL/6 male mice underwent ligation of the left anterior descending coronary artery for 50 minutes followed by reperfusion for 48 hours.In vitro,the primary cultured neonatal mouse ventricular cardiomyocytes(NMVCs)were treated with 100μmol/L hydrogen peroxide(H_(2)O_(2)).The knockdown of lncRNA-AK138945 was evaluated to detect cardiomyocyte apoptosis,and a glucose-regulated,endoplasmic reticulum stress-related protein 94(GRP94)inhibitor was used to detect myocardial injury.Results:We found that the expression level of lncRNA-AK138945 was reduced in MIRI mouse heart tissue and H2O2-treated cardiomyocytes.Moreover,the proportion of apoptosis in cardiomyocytes increased after lncRNA-AK138945 was silenced.The expression level of Bcl2 protein was decreased,and the expression level of Bad,Caspase 9 and Caspase 3 protein was increased.Our further study found that miR-1a-3p is a direct target of lncRNA-AK138945,after lncRNA-AK138945 was silenced in cardiomyocytes,the expression level of miR-1a-3p was increased while the expression level of its downstream protein GRP94 was decreased.Interestingly,treatment with a GRP94 inhibitor(PU-WS13)intensified H2O2-induced cardiomyocyte apoptosis.After overexpression of FOXO3,the expression levels of lncRNA-AK138945 and GRP94 were increased,while the expression levels of miR-1a-3p were decreased.Conclusion:LncRNA-AK138945 inhibits GRP94 expression by regulating miR-1a-3p,leading to cardiomyocyte apoptosis.The transcription factor Forkhead Box Protein O3(FOXO3)participates in cardiomyocyte apoptosis induced by endoplasmic reticulum stress through up-regulation of lncRNA-AK138945.展开更多
目的探讨葡萄糖调节蛋白94(glucose-regulated protein 94,GRP94)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)水平对房颤患者消融术后远期复发事件的预测价值。方法选择因房颤行射频消融术的住院患者共202例,统一归于...目的探讨葡萄糖调节蛋白94(glucose-regulated protein 94,GRP94)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)水平对房颤患者消融术后远期复发事件的预测价值。方法选择因房颤行射频消融术的住院患者共202例,统一归于房颤组,再依据房颤发生及持续的时间进一步分为阵发性房颤组(n=132)和持续性房颤组(n=70);另选择窦性心律且既往无房颤心律的健康体检者作为对照组(n=55)。通过酶联免疫吸附试验测定患者的血清GRP94及TGF-β1水平,并收集基线资料。出院后以门诊或电话形式对房颤患者进行随访,利用受试者工作特征曲线和Kaplan-Meier生存分析评估血清GRP94、TGF-β1水平对房颤患者消融术后远期复发事件的预测价值。结果在对照组、阵发性房颤组、持续性房颤组中,血清GRP94及TGF-β1水平依次升高。二元Logistic回归分析提示,血清TGF-β1、GRP94和体重指数是房颤发生的独立危险因素。Spearman相关性分析显示,在房颤患者血清中GRP94和TGF-β1水平呈正相关。复发组的TGF-β1高于非复发组,GRP94低于非复发组。二元Logistic回归分析表明,TGF-β1是房颤术后复发的独立危险因素,GRP94是房颤复发的独立保护因素。Kaplan-Meier曲线分析显示,GRP94高水平、TGF-β1低水平的房颤患者消融术后复发的风险低,且未复发时间更长。结论GRP94及TGF-β1水平与房颤密切相关,可能为房颤患者消融术后复发的早期识别、诊断及预防提供依据。展开更多
基金This work was supported in part by the National Natural Science Foundation of China(82370417,81970320,82270273)the Certificate of China Postdoctoral Science Foundation Grant(2021M693826)+1 种基金the postdoctoral funding from Heilongjiang Province(21042230046)the Hai Yan Youth Fund from Harbin Medical University Cancer Hospital(JJQN2021-09).
文摘Objective:Myocardial ischemia-reperfusion injury(MIRI)is one of the leading causes of death from cardiovascular disease in humans,especially in individuals exposed to cold environments.Long non-coding RNAs(lncRNAs)regulate MIRI through multiple mechanisms.This study explored the regulatory effect of lncRNA-AK138945 on myocardial ischemia-reperfusion injury and its mechanism.Methods:In vivo,8-to 12-weeks-old C57BL/6 male mice underwent ligation of the left anterior descending coronary artery for 50 minutes followed by reperfusion for 48 hours.In vitro,the primary cultured neonatal mouse ventricular cardiomyocytes(NMVCs)were treated with 100μmol/L hydrogen peroxide(H_(2)O_(2)).The knockdown of lncRNA-AK138945 was evaluated to detect cardiomyocyte apoptosis,and a glucose-regulated,endoplasmic reticulum stress-related protein 94(GRP94)inhibitor was used to detect myocardial injury.Results:We found that the expression level of lncRNA-AK138945 was reduced in MIRI mouse heart tissue and H2O2-treated cardiomyocytes.Moreover,the proportion of apoptosis in cardiomyocytes increased after lncRNA-AK138945 was silenced.The expression level of Bcl2 protein was decreased,and the expression level of Bad,Caspase 9 and Caspase 3 protein was increased.Our further study found that miR-1a-3p is a direct target of lncRNA-AK138945,after lncRNA-AK138945 was silenced in cardiomyocytes,the expression level of miR-1a-3p was increased while the expression level of its downstream protein GRP94 was decreased.Interestingly,treatment with a GRP94 inhibitor(PU-WS13)intensified H2O2-induced cardiomyocyte apoptosis.After overexpression of FOXO3,the expression levels of lncRNA-AK138945 and GRP94 were increased,while the expression levels of miR-1a-3p were decreased.Conclusion:LncRNA-AK138945 inhibits GRP94 expression by regulating miR-1a-3p,leading to cardiomyocyte apoptosis.The transcription factor Forkhead Box Protein O3(FOXO3)participates in cardiomyocyte apoptosis induced by endoplasmic reticulum stress through up-regulation of lncRNA-AK138945.
文摘目的探讨葡萄糖调节蛋白94(glucose-regulated protein 94,GRP94)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)水平对房颤患者消融术后远期复发事件的预测价值。方法选择因房颤行射频消融术的住院患者共202例,统一归于房颤组,再依据房颤发生及持续的时间进一步分为阵发性房颤组(n=132)和持续性房颤组(n=70);另选择窦性心律且既往无房颤心律的健康体检者作为对照组(n=55)。通过酶联免疫吸附试验测定患者的血清GRP94及TGF-β1水平,并收集基线资料。出院后以门诊或电话形式对房颤患者进行随访,利用受试者工作特征曲线和Kaplan-Meier生存分析评估血清GRP94、TGF-β1水平对房颤患者消融术后远期复发事件的预测价值。结果在对照组、阵发性房颤组、持续性房颤组中,血清GRP94及TGF-β1水平依次升高。二元Logistic回归分析提示,血清TGF-β1、GRP94和体重指数是房颤发生的独立危险因素。Spearman相关性分析显示,在房颤患者血清中GRP94和TGF-β1水平呈正相关。复发组的TGF-β1高于非复发组,GRP94低于非复发组。二元Logistic回归分析表明,TGF-β1是房颤术后复发的独立危险因素,GRP94是房颤复发的独立保护因素。Kaplan-Meier曲线分析显示,GRP94高水平、TGF-β1低水平的房颤患者消融术后复发的风险低,且未复发时间更长。结论GRP94及TGF-β1水平与房颤密切相关,可能为房颤患者消融术后复发的早期识别、诊断及预防提供依据。
文摘【背景】随着规模化、集约化生产程度的不断提高,养殖过程中饲养空间受限、冷热环境不适等因素常使猪处于应激状态。内质网应激(endoplasmic reticulum stress,ERS)可能是最早期的应激反应,与细胞凋亡、代谢等方面有密切联系。肝脏是机体的主要代谢器官,猪养殖过程中由于人工操作(如断奶)、饲料霉变、温度变化和吸入有害气体等因素都会造成猪肝脏的ERS,不仅会造成肝脏损伤,还会引发肝脏的脂肪代谢紊乱和广泛的炎症反应,影响生产性能和繁殖性能。因此,深入探讨缓解ERS的有效措施,有助于减少猪养殖过程中的隐性损失。【目的】利用免疫沉淀联合质谱技术,从猪肝星状细胞中筛选在ERS条件下与葡萄糖调节蛋白94(GRP94)相互作用的细胞蛋白,为进一步探讨GRP94对猪肝星状细胞生物学功能的保护作用机理奠定基础。【方法】首先将GRP94抗体固定在谷胱甘肽亲和磁珠上,用亲和磁珠与ERS条件下或正常条件下猪肝星状细胞总蛋白进行孵育,与GRP94诱饵蛋白结合的蛋白复合物洗脱收集后,进行SDS-PAGE凝胶电泳验证。将验证成功的样品洗脱液进行液相色谱串联质谱(LC-MS/MS)检测,鉴定出正常条件和ERS条件下GRP94的互作蛋白。运用生物信息学在线软件对筛选的互作细胞蛋白进行GO富集、KEGG信号通路注释和蛋白互作网络分析,并对其中的互作蛋白之一波形蛋白(vimentin)进行免疫共沉淀验证。【结果】筛选到正常条件下与GRP94存在互作关系的蛋白146个,ERS条件下与GRP94存在互作关系的蛋白76个,在两种情况下都存在互作关系的蛋白44个。ERS条件下有互作关系的76个蛋白质主要参与凋亡过程负调控、肽段交联、泛素依赖型ERAD(endoplasmic reticulum associated degradation)过程和过氧化氢分解代谢等过程。其中参与凋亡过程负调控的GRP94互作蛋白有albumin、catalase、filament A、heat shock protein family A member 5、keratin 18和prohibin 2,说明GRP94可能与这些蛋白共同发挥抗凋亡作用。除此之外组成中间丝纤维的vimentin蛋白参与多个GO富集的通路,可能与GRP94有重要的互作关系。进一步的免疫共沉淀试验也证实,ERS条件下vimentin和GRP94之间确实存在互作关系。此外,某些ERS条件下特异性表达的GRP94互作蛋白(如peroxiredoxin、death inducer obliterator 1、catalase、glandular kallikrein、pyruvate kinase等)与抗凋亡有密切联系。【结论】ERS条件下,猪肝脏GRP94互作蛋白主要参与抗凋亡、对未折叠蛋白进行折叠和维护细胞内稳态相关的信号通路。该结论为下一步开展GRP94参与肝脏ERS调控机制的研究打下基础。
