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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 ROS levels. DCFH-DA staining followed by photofluorography was carried out to observe intracellular ROS levels. Quantitative analysis of the mean fluorescence intensity (MFI) of DCFH-DA with Image-Pro Plus software. Mean ± SEM, n = 4. *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
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Fig6: The effect of exogenous H2S on ROS levels. DCFH-DA staining followed by photofluorography was carried out to observe intracellular ROS levels. Quantitative analysis of the mean fluorescence intensity (MFI) of DCFH-DA with Image-Pro Plus software. Mean ± SEM, n = 4. *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: MDA contents and ROS levels were increased and the activity of SOD, CAT and GSH-PX was decreased in the H/R group (p < 0.05 versus the control group). MDA contents and ROS levels were decreased and the activity of SOD, CAT and GSH-PX was increased in the H/R + NaHS group (p < 0.05 versus the H/R group). The change of these indexes in the PC group was similar to that in the H/R group. Compared with the H/R + NaHS, PC + NaHS futher decreased the content of MDA and the level of ROS, and increased SOD, CAT and GSH-PX activities (p < 0.05). The effect of PC + NaHS on oxidative stress related factors was similar to PC + NAC (or PDTC, or AG 490), respectively (Figs. 5, 6).Fig. 5


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 ROS levels. DCFH-DA staining followed by photofluorography was carried out to observe intracellular ROS levels. Quantitative analysis of the mean fluorescence intensity (MFI) of DCFH-DA with Image-Pro Plus software. Mean ± SEM, n = 4. *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

Fig6: The effect of exogenous H2S on ROS levels. DCFH-DA staining followed by photofluorography was carried out to observe intracellular ROS levels. Quantitative analysis of the mean fluorescence intensity (MFI) of DCFH-DA with Image-Pro Plus software. Mean ± SEM, n = 4. *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: MDA contents and ROS levels were increased and the activity of SOD, CAT and GSH-PX was decreased in the H/R group (p < 0.05 versus the control group). MDA contents and ROS levels were decreased and the activity of SOD, CAT and GSH-PX was increased in the H/R + NaHS group (p < 0.05 versus the H/R group). The change of these indexes in the PC group was similar to that in the H/R group. Compared with the H/R + NaHS, PC + NaHS futher decreased the content of MDA and the level of ROS, and increased SOD, CAT and GSH-PX activities (p < 0.05). The effect of PC + NaHS on oxidative stress related factors was similar to PC + NAC (or PDTC, or AG 490), respectively (Figs. 5, 6).Fig. 5

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