Aims More and more advances show that hydrogen sulfide （H2S） sulfhydrates functional proteins and regulates their actions. However, much less is known about the endogenous sulfhydration of key proteins under physiol...Aims More and more advances show that hydrogen sulfide （H2S） sulfhydrates functional proteins and regulates their actions. However, much less is known about the endogenous sulfhydration of key proteins under physiological conditions. Methods Biotin-switch assay, electrophysiological recording, RNA interference, full ESI-MS scan et al. Results Herein, we found that the protein sulfhydration was dynamically regulated by neuro- nal activity. H2S sulfhydrated and activated serine racemase （SR） in the hippocampus, resuhantly increased D- serine level. Notably, sulfhydration of SR was markedly promoted by neuronal stimulation and underlay activity-de- pendent changes of D-serine availability. Both genetic knockdown and pharmacological inhibition of cystathionine β-synthase （CBS）, the key H2S-producing enzyme in brain, attenuated hippoeampal long-term potentiation （LTP） , which can be reversed by exogenous supplement of H2S or D-serine. We also observed that the increase in LTP induced by H2S is dependent on D-serine and polysulfides. In aged rats, sulfhydration of SR was significantly decreased. Furthermore, exogenous supplement of H2S restored the level of sulfhydration and reversed the age-re- lated deficits in hippocampal LTP. Conclusion Our data provide direct evidence for the biological significance of endogenous sulfhydration in physiological conditions and demonstrate a pivotal role of activity-dependent sulfhydra- tion in synaptic plasticity.展开更多
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
文摘Aims More and more advances show that hydrogen sulfide （H2S） sulfhydrates functional proteins and regulates their actions. However, much less is known about the endogenous sulfhydration of key proteins under physiological conditions. Methods Biotin-switch assay, electrophysiological recording, RNA interference, full ESI-MS scan et al. Results Herein, we found that the protein sulfhydration was dynamically regulated by neuro- nal activity. H2S sulfhydrated and activated serine racemase （SR） in the hippocampus, resuhantly increased D- serine level. Notably, sulfhydration of SR was markedly promoted by neuronal stimulation and underlay activity-de- pendent changes of D-serine availability. Both genetic knockdown and pharmacological inhibition of cystathionine β-synthase （CBS）, the key H2S-producing enzyme in brain, attenuated hippoeampal long-term potentiation （LTP） , which can be reversed by exogenous supplement of H2S or D-serine. We also observed that the increase in LTP induced by H2S is dependent on D-serine and polysulfides. In aged rats, sulfhydration of SR was significantly decreased. Furthermore, exogenous supplement of H2S restored the level of sulfhydration and reversed the age-re- lated deficits in hippocampal LTP. Conclusion Our data provide direct evidence for the biological significance of endogenous sulfhydration in physiological conditions and demonstrate a pivotal role of activity-dependent sulfhydra- tion in synaptic plasticity.
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.
