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
钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)是一种在学习和记忆形成机制中具有重要作用的蛋白激酶,它存在于大多数组织中,但以神经元中表达量最高。大多数具有激酶活性的蛋白分子在组织中的表达量...钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)是一种在学习和记忆形成机制中具有重要作用的蛋白激酶,它存在于大多数组织中,但以神经元中表达量最高。大多数具有激酶活性的蛋白分子在组织中的表达量都相对较低,所以CaMKⅡ在神经系统中高度表达具有其特殊意义。该文将较为全面的对CaMKⅡ的分子结构与自身磷酸化特征、亚细胞空间定位及其在突触可塑性中的作用进行综述。展开更多
CaMKⅡ是钙/钙调蛋白激酶(Ca2+/calmodulin-dependent protein kinase,CaMK)成员之一。心脏中的CaMK包括Ⅰ,Ⅱ和Ⅳ三种类型,CaMKⅡ含量最多。CaMKⅡ单体由氨基端的催化域、中间部分的调节域和羧基端的结合域组成。钙调蛋白(calmodulin,C...CaMKⅡ是钙/钙调蛋白激酶(Ca2+/calmodulin-dependent protein kinase,CaMK)成员之一。心脏中的CaMK包括Ⅰ,Ⅱ和Ⅳ三种类型,CaMKⅡ含量最多。CaMKⅡ单体由氨基端的催化域、中间部分的调节域和羧基端的结合域组成。钙调蛋白(calmodulin,CaM)与Ca2+结合后被激活,结合于CaMKⅡ调节域中的CaM结合区激活CaMKⅡ。CaMKⅡ富集于T管并靠近L-Ca2+通道,也存在于肌浆网和细胞核中。激活的CaMKⅡ通过多种途径调节细胞内Ca2+平衡,广泛参与心血管系统生理活动及病理变化的信号转导过程,与多种心血管系统疾病密切相关。目前研究认为,CaMKⅡ信号系统在心律失常、心肌肥厚、心力衰竭、缺血性心脏病和扩张性心肌病的发生和发展中起着重要作用。展开更多
OBJECTIVE Dorsal raphe nucleus(DRN) is the largest single collection of neurons containing5-HT in the entire brain and particularly attractive in a wide variety of complex physiological and behavioral processes,such a...OBJECTIVE Dorsal raphe nucleus(DRN) is the largest single collection of neurons containing5-HT in the entire brain and particularly attractive in a wide variety of complex physiological and behavioral processes,such as sleep-wake regulation. Calmodulin dependent kinaseⅡ(CaMKⅡ) and protein kinase C(PKC)are important signal-transducing molecules activated by Ca^(2+). Since the Ca^(2+)modulation in DRN plays an important role in sleep-wake regulation,it should be presumed that the intracellular CaMKⅡ/PKC signaling in DRN may be involved in the regulation of sleep-wake. METHODS The polysomnogram consisting of EEG and EMG was recorded for analyzing sleep architecture. Immunohistochemisrty and Western-blotting methods were used in this study to investigate the roles of Ca^(2+),CaMKⅡ and PKC in sleep-wake regulation in rat DRN. RESULTS Ca^(2+)in the DRN exert arousal effects by reducing the NREMs,SWS and REMs via up-regulating serotonergic functions and activating CaMK Ⅱ-PKC.However,inhibition of PKC leads to significant promotion of total sleep time especial y the NREM sleep,but there were no changes in sleep parameters after the inhibition of CaMKⅡ by its inhibitor KN-93 in DRN.CONCLUSION The molecular,pharmacological,and behavioral findings of this study demonstrate a novel wake promoting and sleep-suppressing role for the Ca^(2+)/CaMK Ⅱ/PKC signaling pathway in DRN. Abnormalities in CaMK Ⅱ are found in patients with several neurological disorders that are associated with disturbed sleep,such as schizophrenia,depression,and Alzheimer′s disease. Several psychotropic drugs modulate CaMK Ⅱ activity. In addition,PKC is a cellular target of most current mood stabilizing and anti-manic agents and involved in bipolar disorder. The data of the present study raise the question whether PKC or CaMKⅡ modulations may also be effective on the sleep disorders or the mood disorders associated with sleep disorders.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.
文摘钙调素依赖性蛋白激酶Ⅱ(calcium/calmodulin-dependent protein kinaseⅡ,CaMKⅡ)是一种在学习和记忆形成机制中具有重要作用的蛋白激酶,它存在于大多数组织中,但以神经元中表达量最高。大多数具有激酶活性的蛋白分子在组织中的表达量都相对较低,所以CaMKⅡ在神经系统中高度表达具有其特殊意义。该文将较为全面的对CaMKⅡ的分子结构与自身磷酸化特征、亚细胞空间定位及其在突触可塑性中的作用进行综述。
文摘CaMKⅡ是钙/钙调蛋白激酶(Ca2+/calmodulin-dependent protein kinase,CaMK)成员之一。心脏中的CaMK包括Ⅰ,Ⅱ和Ⅳ三种类型,CaMKⅡ含量最多。CaMKⅡ单体由氨基端的催化域、中间部分的调节域和羧基端的结合域组成。钙调蛋白(calmodulin,CaM)与Ca2+结合后被激活,结合于CaMKⅡ调节域中的CaM结合区激活CaMKⅡ。CaMKⅡ富集于T管并靠近L-Ca2+通道,也存在于肌浆网和细胞核中。激活的CaMKⅡ通过多种途径调节细胞内Ca2+平衡,广泛参与心血管系统生理活动及病理变化的信号转导过程,与多种心血管系统疾病密切相关。目前研究认为,CaMKⅡ信号系统在心律失常、心肌肥厚、心力衰竭、缺血性心脏病和扩张性心肌病的发生和发展中起着重要作用。
基金The project supported by National Natural Science Foundation of China(81573407,81302746,81202511,81173031)National Mega-project of Scicence Research of China for New Drug Development(2009ZX09103-124)Research Fund for the Doctoral Program of Higher Eductaion of China(20100001110048)
文摘OBJECTIVE Dorsal raphe nucleus(DRN) is the largest single collection of neurons containing5-HT in the entire brain and particularly attractive in a wide variety of complex physiological and behavioral processes,such as sleep-wake regulation. Calmodulin dependent kinaseⅡ(CaMKⅡ) and protein kinase C(PKC)are important signal-transducing molecules activated by Ca^(2+). Since the Ca^(2+)modulation in DRN plays an important role in sleep-wake regulation,it should be presumed that the intracellular CaMKⅡ/PKC signaling in DRN may be involved in the regulation of sleep-wake. METHODS The polysomnogram consisting of EEG and EMG was recorded for analyzing sleep architecture. Immunohistochemisrty and Western-blotting methods were used in this study to investigate the roles of Ca^(2+),CaMKⅡ and PKC in sleep-wake regulation in rat DRN. RESULTS Ca^(2+)in the DRN exert arousal effects by reducing the NREMs,SWS and REMs via up-regulating serotonergic functions and activating CaMK Ⅱ-PKC.However,inhibition of PKC leads to significant promotion of total sleep time especial y the NREM sleep,but there were no changes in sleep parameters after the inhibition of CaMKⅡ by its inhibitor KN-93 in DRN.CONCLUSION The molecular,pharmacological,and behavioral findings of this study demonstrate a novel wake promoting and sleep-suppressing role for the Ca^(2+)/CaMK Ⅱ/PKC signaling pathway in DRN. Abnormalities in CaMK Ⅱ are found in patients with several neurological disorders that are associated with disturbed sleep,such as schizophrenia,depression,and Alzheimer′s disease. Several psychotropic drugs modulate CaMK Ⅱ activity. In addition,PKC is a cellular target of most current mood stabilizing and anti-manic agents and involved in bipolar disorder. The data of the present study raise the question whether PKC or CaMKⅡ modulations may also be effective on the sleep disorders or the mood disorders associated with sleep disorders.
