Limits...
Exogenous H2S contributes to recovery of ischemic post-conditioning-induced cardioprotection by decrease of ROS level via down-regulation of NF-κB and JAK2-STAT3 pathways in the aging cardiomyocytes.

Li L, Li M, Li Y, Sun W, Wang Y, Bai S, Li H, Wu B, Yang G, Wang R, Wu L, Li H, Xu C - Cell Biosci (2016)

Bottom Line: However, PC protection is ineffective in the aging cardiomyocytes.PC alone did not provide cardioprotection in H/R-treated aging cardiomyocytes, which was significantly restored by the addition of NaHS.Our results suggest that exogenous H2S contributes to recovery of PC-induced cardioprotection by decrease of ROS level via down-regulation of NF-κB and JAK2/STAT3 pathways in the aging cardiomyocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology, Harbin Medical University, Baojian Road, Harbin, 150081 China ; The Key Laboratory of Cardiovascular Medicine Research, Harbin Medical University, Ministry of Education, Harbin, China.

ABSTRACT

Background: Hydrogen sulfide (H2S), a third member of gasotransmitter family along with nitric oxide and carbon monoxide, generated from mainly catalyzed by cystathionine-lyase, possesses important functions in the cardiovascular system. Ischemic post-conditioning (PC) strongly protects against the hypoxia/reoxygenation (H/R)-induced injury and apoptosis of cardiomyocytes. However, PC protection is ineffective in the aging cardiomyocytes. Whether H2S restores PC-induced cardioprotection by decrease of reactive oxygen species (ROS) level in the aging cardiomyocytes is unknown.

Methods: The aging cardiomyocytes were induced by treatment of primary cultures of neonatal cardiomyocytes using d-galactose and were exposed to H/R and PC protocols. Cell viability was observed by CCK-8 kit. Apoptosis was detected by Hoechst 33342 staining and flow cytometry. ROS level was analyzed using spectrofluorimeter. Related protein expressions were detected through Western blot.

Results: Treatment of NaHS (a H2S donor) protected against H/R-induced apoptosis, cell damage, the expression of cleaved caspase-3 and cleaved caspase-9, the release of cytochrome c (Cyt c). The supplementation of NaHS also decreased the activity of LDH and CK, MDA contents, ROS levels and the phosphorylation of IκBα, NF-κB, JNK2 and STAT3, and increased cell viability, the expression of Bcl-2, the activity of SOD, CAT and GSH-PX. PC alone did not provide cardioprotection in H/R-treated aging cardiomyocytes, which was significantly restored by the addition of NaHS. The beneficial role of NaHS was similar to the supply of N-acetyl-cysteine (NAC, an inhibitor of ROS), Ammonium pyrrolidinedithiocarbamate (PDTC, an inhibitor of NF-κB) and AG 490 (an inhibitor of JNK2), respectively, during PC.

Conclusion: Our results suggest that exogenous H2S contributes to recovery of PC-induced cardioprotection by decrease of ROS level via down-regulation of NF-κB and JAK2/STAT3 pathways in the aging cardiomyocytes.

No MeSH data available.


Related in: MedlinePlus

The effect of exogenous H2S on the NF-κB pathway. The phosphorylation of IκBα and NF-κB was detected using Western bloting. The graphs represent the optical density of the bands of phosphorylated IκBα and NF-κB normalized with the expression of total IκBα and NF-κB, respectively. All data were from three independent experiments. *p < 0.05 vs. control group; #p < 0.05 vs. H/R group; &p < 0.05 vs. PC group; $p < 0.05 vs. H/R + NaHS group
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4836181&req=5

Fig7: The effect of exogenous H2S on the NF-κB pathway. The phosphorylation of IκBα and NF-κB was detected using Western bloting. The graphs represent the optical density of the bands of phosphorylated IκBα and NF-κB normalized with the expression of total IκBα and NF-κB, respectively. All data were from three independent experiments. *p < 0.05 vs. control group; #p < 0.05 vs. H/R group; &p < 0.05 vs. PC group; $p < 0.05 vs. H/R + NaHS group

Mentions: The activity of phosphorylated IκBα and NF-κB in the H/R group was significantly higher than that in the control group (p < 0.05). The activity of phosphorylated IκBα and NF-κB was markedly lower in the H/R + NaHS group than in the H/R group (p < 0.05). The change of phosphorylated IκBα and NF-κB activities in the PC group were similar to those in the H/R group. In the PC + NaHS group, the activity of phosphorylated IκBα and NF-κB was decreased to a larger extent than that in the H/R + NaHS (p < 0.05). The effect of PC + NaHS on NF-κB pathway was similar to PC + NAC or PC + PDTC, respectively. The total amount of IκBα and NF-κB protein remained unchanged with the different stimulations (Fig. 7).Fig. 7


Exogenous H2S contributes to recovery of ischemic post-conditioning-induced cardioprotection by decrease of ROS level via down-regulation of NF-κB and JAK2-STAT3 pathways in the aging cardiomyocytes.

Li L, Li M, Li Y, Sun W, Wang Y, Bai S, Li H, Wu B, Yang G, Wang R, Wu L, Li H, Xu C - Cell Biosci (2016)

The effect of exogenous H2S on the NF-κB pathway. The phosphorylation of IκBα and NF-κB was detected using Western bloting. The graphs represent the optical density of the bands of phosphorylated IκBα and NF-κB normalized with the expression of total IκBα and NF-κB, respectively. All data were from three independent experiments. *p < 0.05 vs. control group; #p < 0.05 vs. H/R group; &p < 0.05 vs. PC group; $p < 0.05 vs. H/R + NaHS group
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4836181&req=5

Fig7: The effect of exogenous H2S on the NF-κB pathway. The phosphorylation of IκBα and NF-κB was detected using Western bloting. The graphs represent the optical density of the bands of phosphorylated IκBα and NF-κB normalized with the expression of total IκBα and NF-κB, respectively. All data were from three independent experiments. *p < 0.05 vs. control group; #p < 0.05 vs. H/R group; &p < 0.05 vs. PC group; $p < 0.05 vs. H/R + NaHS group
Mentions: The activity of phosphorylated IκBα and NF-κB in the H/R group was significantly higher than that in the control group (p < 0.05). The activity of phosphorylated IκBα and NF-κB was markedly lower in the H/R + NaHS group than in the H/R group (p < 0.05). The change of phosphorylated IκBα and NF-κB activities in the PC group were similar to those in the H/R group. In the PC + NaHS group, the activity of phosphorylated IκBα and NF-κB was decreased to a larger extent than that in the H/R + NaHS (p < 0.05). The effect of PC + NaHS on NF-κB pathway was similar to PC + NAC or PC + PDTC, respectively. The total amount of IκBα and NF-κB protein remained unchanged with the different stimulations (Fig. 7).Fig. 7

Bottom Line: However, PC protection is ineffective in the aging cardiomyocytes.PC alone did not provide cardioprotection in H/R-treated aging cardiomyocytes, which was significantly restored by the addition of NaHS.Our results suggest that exogenous H2S contributes to recovery of PC-induced cardioprotection by decrease of ROS level via down-regulation of NF-κB and JAK2/STAT3 pathways in the aging cardiomyocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology, Harbin Medical University, Baojian Road, Harbin, 150081 China ; The Key Laboratory of Cardiovascular Medicine Research, Harbin Medical University, Ministry of Education, Harbin, China.

ABSTRACT

Background: Hydrogen sulfide (H2S), a third member of gasotransmitter family along with nitric oxide and carbon monoxide, generated from mainly catalyzed by cystathionine-lyase, possesses important functions in the cardiovascular system. Ischemic post-conditioning (PC) strongly protects against the hypoxia/reoxygenation (H/R)-induced injury and apoptosis of cardiomyocytes. However, PC protection is ineffective in the aging cardiomyocytes. Whether H2S restores PC-induced cardioprotection by decrease of reactive oxygen species (ROS) level in the aging cardiomyocytes is unknown.

Methods: The aging cardiomyocytes were induced by treatment of primary cultures of neonatal cardiomyocytes using d-galactose and were exposed to H/R and PC protocols. Cell viability was observed by CCK-8 kit. Apoptosis was detected by Hoechst 33342 staining and flow cytometry. ROS level was analyzed using spectrofluorimeter. Related protein expressions were detected through Western blot.

Results: Treatment of NaHS (a H2S donor) protected against H/R-induced apoptosis, cell damage, the expression of cleaved caspase-3 and cleaved caspase-9, the release of cytochrome c (Cyt c). The supplementation of NaHS also decreased the activity of LDH and CK, MDA contents, ROS levels and the phosphorylation of IκBα, NF-κB, JNK2 and STAT3, and increased cell viability, the expression of Bcl-2, the activity of SOD, CAT and GSH-PX. PC alone did not provide cardioprotection in H/R-treated aging cardiomyocytes, which was significantly restored by the addition of NaHS. The beneficial role of NaHS was similar to the supply of N-acetyl-cysteine (NAC, an inhibitor of ROS), Ammonium pyrrolidinedithiocarbamate (PDTC, an inhibitor of NF-κB) and AG 490 (an inhibitor of JNK2), respectively, during PC.

Conclusion: Our results suggest that exogenous H2S contributes to recovery of PC-induced cardioprotection by decrease of ROS level via down-regulation of NF-κB and JAK2/STAT3 pathways in the aging cardiomyocytes.

No MeSH data available.


Related in: MedlinePlus