Exogenous Hydrogen Sulfide Protects against Doxorubicin-Induced Inflammation and Cytotoxicity by Inhibiting p38MAPK/NFκB Pathway in H9c2 Cardiac Cells

Abstract

Background/Aim:We have demonstrated that exogenous hydrogen sulfide (H₂S) protects H9c2 cardiac cells against the doxorubicin (DOX)-induced injuries by inhibiting p38 mitogen-activated protein kinase (MAPK) pathway and that the p38 MAPK/nuclear factor-κB (NF-κB) pathway is involved in the DOX-induced inflammatory response and cytotoxicity. The present study attempts to test the hypothesis that exogenous H₂S might protect cardiomyocytes against the DOX-induced inflammation and cytotoxicity through inhibiting p38 MAPK/NF-κB pathway. Methods: H9c2 cardiac cells were exposed to 5µM DOX for 24 h to establish a model of DOX cardiotoxicity. The cells were pretreated with NaHS( a donor of H₂S) or other drugs before exposure to DOX. Cell viability was analyzed by cell counter kit 8 ( CCK-8), The expression of NF-κB p65 and inducible nitric oxide synthase (iNOS) was detected by Western blot assay. The levels of interleukin-1ß (IL-1ß), IL-6 and tumor necrosis factor-a (TNF-a) were tested by enzyme-linked immunosorbent assay (ELISA). Results: Our findings demonstrated that pretreatment of H9c2 cardiac cells with NaHS for 30 min before exposure to DOX markedly ameliorated the DOX-induced phosphorylation and nuclear translocation of NF-κB p65 subunit. Importantly, the pretreatment with NaHS significantly attenuated the p38 MAPK/NF-κB pathway-mediated inflammatory responses induced by DOX, as evidenced by decreases in the levels of IL-1ß, IL-6 and TNF-a. In addition, application of NaHS or IL-1ß receptor antagonist (IL-1Ra) or PDTC (an inhibitor of NF-κB) attenuated the DOX-induced expression of iNOS and production of nitric oxide (NO), respectively. Furthermore, IL-1Ra also dramatically reduced the DOX-induced cytotoxicity and phosphorylation of NF-κB p65. The pretreatment of H9c2 cells with N-acetyl-L-cysteine (NAC), a scavenger of reactive oxygen species (ROS) prior to exposure to DOX depressed the phosphorylation of NF-κB p65 induced by DOX. Conclusion: The present study has demonstrated the new mechanistic evidence that exogenous H₂S attenuates the DOX-induced inflammation and cytotoxicity by inhibiting p38 MAPK/NF-κB pathway in H9c2 cardiac cells. We also provide novel data that the interaction between NF-κB pathway and IL-1ß is important in the induction of DOX-induced inflammation and cytotoxicity in H9c2 cardiac cells.

Resveratrol inhibits doxorubicin-induced cardiotoxicity via sirtuin 1 activation in H9c2 cardiomyocytes

“…Numerous studies have demonstrated that the primary molecular mechanism involved in DOX-induced cardiotoxicity is free radical-induced oxidative stress, and cardiomyocyte death by apoptosis and necrosis (5,22). In accordance with previous studies (23,24), in the present study it was observed that the exposure of H9c2 cells to DOX significantly induced cellular injuries, including decreases in cell viability and the expression of SIRT1, as well as increases in cell apoptosis and in the expression levels of FoxO1, P53 and Bim.…”

Cardioprotection by H2S Donors: Nitric Oxide-Dependent and -Independent Mechanisms

“…H 2 S donors, including diallyl disulfide and S-propargylcysteine, have been shown to rescue cultured cardiomyocytes from high glycose injury, doxorubicin-induced toxicity or ROStriggered injury (Szabó et al, 2011;Guo et al, 2013;Wu et al, 2015;Yang et al, 2015). The mechanisms through which H 2 S donors have been proposed to exerts their protective effects in these cultured cell systems include ATP-sensitive potassium channel and Akt activation, inhibition of mitogen-activated protein kinases pathways (p38, c-Jun N-terminal protein kinases), and inhibition of endoplasmic reticulum stress, as well as antioxidant mechanisms (Szabó et al, 2011;Wang, 2012;Guo et al, 2013;Salloum, 2015;Wu et al, 2015;Yang Fig.…”

“…The mechanisms through which H 2 S donors have been proposed to exerts their protective effects in these cultured cell systems include ATP-sensitive potassium channel and Akt activation, inhibition of mitogen-activated protein kinases pathways (p38, c-Jun N-terminal protein kinases), and inhibition of endoplasmic reticulum stress, as well as antioxidant mechanisms (Szabó et al, 2011;Wang, 2012;Guo et al, 2013;Salloum, 2015;Wu et al, 2015;Yang Fig. 6.…”

Emerging role of hydrogen sulfide-microRNA crosstalk in cardiovascular diseases

“…Members of the MAPK family, p38-MAPK and ERK1/2, are upregulated in response to cellular stress, leading to cardiomyocyte apoptosis (50,156). p38-MAPK is induced in several disease conditions including hyperglycemia, I/R injury, and hypoxia (16,49,50,134,156).…”

“…p38-MAPK is induced in several disease conditions including hyperglycemia, I/R injury, and hypoxia (16,49,50,134,156). NaHS (400 M) inhibits the activation of p38-MAPK in response to the antitumor drug doxorubicin, resulting in decreased apoptosis (50).…”