OBJECTIVE To determine the functional role of hydrogen sulfide(H_2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca^(2+)/calmodulin-dependent protein kinaseⅡ(Ca MKⅡ) us...OBJECTIVE To determine the functional role of hydrogen sulfide(H_2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca^(2+)/calmodulin-dependent protein kinaseⅡ(Ca MKⅡ) using wild type and CSE knockout mouse models.METHODS Continuous subcutaneous injection isoprenaline(7.5 mg·kg^(-1) per day),once a day for 4 weeks to induce heart failure in male C57BL/6(6-8 weeks old) mice and CSE-/-mice.150 μmol·L^(-1) H_2O_2 was used to induce oxidative stress in H9c2 cells.Echocardiograph was used to detect cardiac parameters.H&E stain and Masson stain was to observation histopathological changes.Western blot was used to detect protein expression and activity.The si RNA was used to silence protein expression.HPLC was used to detect H_2S level.Biotin assay was used to detect the level of S-sulfhydration protein.RESULTS Treatment with S-propyl-L-cysteine(SPRC) or sodium hydrosulfide(Na HS),modulators of blood H_2S levels,attenuated the development of heart failure in animals,reduced lipid peroxidation,and preserved mitochondrial function.The inhibition Ca MKⅡ phosphorylation by SPRC and Na HS as demonstrated using both in vivo and in vitro models corresponded with the cardioprotective effects of these compounds.Interestingly,Ca MKⅡ activity was found to be elevated in CSE-/-mice as compared to wild type animals and the phosphorylation status of Ca MK Ⅱ appeared to relate to the severity of heart failure.Importantly,in wild type mice SPRC was found to promote S-sulfhydration of Ca MKⅡ leading to reduced activity of this protein however,in CSE-/-mice S-sulfhydration was abolished following SPRC treatment.CONCLUSION A novel mechanism depicting a role of S-sulfhydration in the regulation of Ca MKⅡ is presented.SPRC mediated S-sulfhydration of Ca MKⅡ was found to inhibit Ca MKⅡ activity and to preserve cardiovascular homeostasis.展开更多
为探究钙调控因子在硬骨鱼类钙代谢过程中发挥的作用,本研究选取细胞内Ca^(2+)信号传导的关键调控因子-钙调素(Calmodulin,CaM)为研究对象,通过对虹鳟(Oncorhynchus mykiss)幼鱼腹腔注射氯化卡咪唑铵(Calmidazolium chloride,CMZ)抑制Ca...为探究钙调控因子在硬骨鱼类钙代谢过程中发挥的作用,本研究选取细胞内Ca^(2+)信号传导的关键调控因子-钙调素(Calmodulin,CaM)为研究对象,通过对虹鳟(Oncorhynchus mykiss)幼鱼腹腔注射氯化卡咪唑铵(Calmidazolium chloride,CMZ)抑制CaM活性,并在注射后6 h采集实验组及对照组(腹腔注射同体积生理盐水)虹鳟鳞片组织,利用转录组测序技术分析实验组mRNA的表达变化。结果显示:转录组测序共获得376.31 M Raw reads,Q30碱基百分比均在93.26%以上;实验组鳞片组织中共筛选出3730个差异表达基因(DEGs),其中1227个基因表达上调,2503个基因表达下调。GO注释结果显示,上述DEGs主要富集到细胞过程、生物调控、细胞器、结合和生物过程调控等功能;KEGG通路富集分析结果显示,DEGs显著富集的前3条通路依次为核糖体通路、Rap1信号通路和轴突导向通路,此外,DEGs还显著富集于破骨细胞分化、TGF-beta等与骨骼发育、钙代谢等密切相关的信号通路。本研究所获得的差异表达基因信息及其功能注释和通路富集结果可为阐明鱼类钙代谢的调控机理提供新素材。展开更多
文摘OBJECTIVE To determine the functional role of hydrogen sulfide(H_2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca^(2+)/calmodulin-dependent protein kinaseⅡ(Ca MKⅡ) using wild type and CSE knockout mouse models.METHODS Continuous subcutaneous injection isoprenaline(7.5 mg·kg^(-1) per day),once a day for 4 weeks to induce heart failure in male C57BL/6(6-8 weeks old) mice and CSE-/-mice.150 μmol·L^(-1) H_2O_2 was used to induce oxidative stress in H9c2 cells.Echocardiograph was used to detect cardiac parameters.H&E stain and Masson stain was to observation histopathological changes.Western blot was used to detect protein expression and activity.The si RNA was used to silence protein expression.HPLC was used to detect H_2S level.Biotin assay was used to detect the level of S-sulfhydration protein.RESULTS Treatment with S-propyl-L-cysteine(SPRC) or sodium hydrosulfide(Na HS),modulators of blood H_2S levels,attenuated the development of heart failure in animals,reduced lipid peroxidation,and preserved mitochondrial function.The inhibition Ca MKⅡ phosphorylation by SPRC and Na HS as demonstrated using both in vivo and in vitro models corresponded with the cardioprotective effects of these compounds.Interestingly,Ca MKⅡ activity was found to be elevated in CSE-/-mice as compared to wild type animals and the phosphorylation status of Ca MK Ⅱ appeared to relate to the severity of heart failure.Importantly,in wild type mice SPRC was found to promote S-sulfhydration of Ca MKⅡ leading to reduced activity of this protein however,in CSE-/-mice S-sulfhydration was abolished following SPRC treatment.CONCLUSION A novel mechanism depicting a role of S-sulfhydration in the regulation of Ca MKⅡ is presented.SPRC mediated S-sulfhydration of Ca MKⅡ was found to inhibit Ca MKⅡ activity and to preserve cardiovascular homeostasis.
文摘为探究钙调控因子在硬骨鱼类钙代谢过程中发挥的作用,本研究选取细胞内Ca^(2+)信号传导的关键调控因子-钙调素(Calmodulin,CaM)为研究对象,通过对虹鳟(Oncorhynchus mykiss)幼鱼腹腔注射氯化卡咪唑铵(Calmidazolium chloride,CMZ)抑制CaM活性,并在注射后6 h采集实验组及对照组(腹腔注射同体积生理盐水)虹鳟鳞片组织,利用转录组测序技术分析实验组mRNA的表达变化。结果显示:转录组测序共获得376.31 M Raw reads,Q30碱基百分比均在93.26%以上;实验组鳞片组织中共筛选出3730个差异表达基因(DEGs),其中1227个基因表达上调,2503个基因表达下调。GO注释结果显示,上述DEGs主要富集到细胞过程、生物调控、细胞器、结合和生物过程调控等功能;KEGG通路富集分析结果显示,DEGs显著富集的前3条通路依次为核糖体通路、Rap1信号通路和轴突导向通路,此外,DEGs还显著富集于破骨细胞分化、TGF-beta等与骨骼发育、钙代谢等密切相关的信号通路。本研究所获得的差异表达基因信息及其功能注释和通路富集结果可为阐明鱼类钙代谢的调控机理提供新素材。
