Sulforaphane prevents doxorubicin-induced oxidative stress and cell death in rat H9c2 cells

Abstract

Sulforaphane, a natural isothiocyanate compound found in cruciferous vegetables, has been shown to exert cardioprotective effects during ischemic heart injury. However, the effects of sulforaphane on cardiotoxicity induced by doxorubicin are unknown. Thus, in the present study, H9c2 rat myoblasts were pre-treated with sulforaphane and its effects on cardiotoxicity were then examined. The results revealed that the pre-treatment of H9c2 rat myoblasts with sulforaphane decreased the apoptotic cell number (as shown by trypan blue exclusion assay) and the expression of pro-apoptotic proteins (Bax, caspase-3 and cytochrome c; as shown by western blot analysis and immunostaining), as well as the doxorubicin-induced increase in mitochondrial membrane potential (measured by JC-1 assay). Furthermore, sulforaphane increased the mRNA and protein expression of heme oxygenase-1 (HO-1, measured by RT-qPCR), which consequently reduced the levels of reactive oxygen species (ROS, measured using MitoSOX Red reagent) in the mitochondria which were induced by doxorubicin. The cardioprotective effects of sulforaphane were found to be mediated by the activation of the Kelch-like ECH-associated protein 1 (Keap1)/NF-E2-related factor-2 (Nrf2)/antioxidant-responsive element (ARE) pathway, which in turn mediates the induction of HO-1. Taken together, the findings of this study demonstrate that sulforaphane prevents doxorubicin-induced oxidative stress and cell death in H9c2 cells through the induction of HO-1 expression.

The natural compound 2,3,5,4′‐tetrahydroxystilbene‐2‐O‐β‐d glucoside protects against adriamycin‐induced nephropathy through activating the Nrf2‐Keap1 antioxidant pathway

“…Evidence indicated that inhibition of Nrf2 by either genetic manipulation or an inhibitor can increase the sensitivity of ovarian and hepatocellular carcinoma cell lines to AD . Although it remains unclear whether Nrf2 also participates in the action of AD‐mediated topoisomerase II activity dysregulation, other and our data all suggest that activation of Nrf2 can suppress AD‐induced cytotoxicity.…”

Gp130-mediated STAT3 activation by S-propargyl-cysteine, an endogenous hydrogen sulfide initiator, prevents doxorubicin-induced cardiotoxicity

“…31 As shown in Figure 5a, Dox caused an approximately 2.7-fold increase in intracellular ROS generation as monitored by DCF fluorescence, which was significantly decreased by SPRC ( P <0.01). Moreover, consistent with recent studies, 31, 32 Dox-treated cells revealed the release of proapoptotic mitochondrial protein cytochrome c into cytosol. In contrast, SPRC prevented Dox-induced cytochrome c release ( P <0.05; Figure 5b).…”

“…45 The release of cytochrome c from mitochondria to cytosol, and dissipation of ΔΨm , is linked to Dox-mediated apoptotic signaling and mitochondrial dysfunction. 31 On the other hand, oxidative stress through cumulative Ca 2+ and mitochondrial dysfunction causes depletion in ATP necessary in the process of myocardial fiber contractions. 46, 47 It can also be a basis for pathological changes in Ca 2+ regulation through SERCA2.…”

“…JC-1 stains the mitochondria in cells with a high ΔΨm by forming red fluorescence J-aggregates, whereas in cells with depolarized mitochondria, JC-1 is present as green fluorescent monomers. 31 In this way, mitochondrial depolarization was determined by a decreased ratio of red-to-green fluorescence intensity by a fluorescence microscope (Zeiss Inc.).…”

Protective effects of mito-TEMPO against doxorubicin cardiotoxicity in mice

“…Although DOX-induced cardiac toxicity appears to be multifactorial, the most thoroughly investigated hypothesis has been the formation of reactive oxygen species (ROS) and there is evidence pointing to cardiac mitochondria as primary targets of the toxicity of DOX [3]. The quinone moiety of DOX may form semiquinone radicals by oneelectron reduction.…”