Thymoquinone‑induced autophagy mitigates doxorubicin‑induced H9c2 cell apoptosis

Abstract

Doxorubicin (Dox)-induced myocardiopathy is a massive obstacle in administering chemotherapeutic drugs in cancer patients. In the present study, the effects of thymoquinone (TQ) on Dox-induced cardiotoxicity were investigated in vitro. H9c2 cardiomyocytes were pre-treated with TQ followed by Dox, and cell viability and cell death were subsequently measured. Flow cytometry was conducted to evaluate the apoptosis of H9c2 cells. Autophagy was assessed by immunofluorescence LC3 staining. Western blot analysis was performed to determine autophagy-, apoptosisand LKB1/AMPK-related proteins. Dox-induced H9c2 cell death and apoptosis were decreased after pretreatment with TQ in vitro. The number of autophagosomes and autophagic markers was further increased by TQ in the Dox-treated H9c2 cells, which mediated LKB1/AMPK activation and the deactivation of mTOR. Suppression of autophagosomes abolished the TQ-induced antiapoptotic effect. In conclusion, TQ reduces Dox-induced apoptosis of cardiomyocytes by upregulating autophagy through activation of the LKB1/AMPK pathways. These novel results highlight the therapeutic potential of TQ to prevent Dox cardiotoxicity.

Protective effect of thymoquinone nanoemulsion in reducing the cardiotoxic effect of 5‐fluorouracil in rats

“…Also, other studies have shown the protective effects of TMQ in reducing cardiotoxicity (Karabulut et al, 2021; Öztürk et al, 2020). In the study by Liu et al, treatment with TMQ significantly reduced cardiotoxicity caused by doxorubicin (Liu & Zhao, 2022).…”

Effects of extracellular vesicles from adipose‐derived stem cells on human keloid fibroblasts via the SOCS1/JAK2/STAT3 pathway

“…As light chain 3 (LC3)‐II serves as a widely accepted marker for autophagosomes and relatively specifically related to autophagosomes and autolysosomes, the quantification of LC3‐positive puncta is deemed gold‐standard for determining autophagosome numbers in cells 37 . Immunofluorescence assay was performed as reported 38 . HKFs were fixed for 15 min with 4% paraformaldehyde, rinsed with NaCl/Pi, permeabilized with 0.2% Triton X‐100, and incubated at 4°C with LC3B primary rabbit antibody (1:200, #3868, CST, Danvers, MA, USA) overnight.…”

“…37 Immunofluorescence assay was performed as reported. 38 HKFs were fixed for 15 min with 4% paraformaldehyde, rinsed with NaCl/Pi, permeabilized with 0.2% Triton X-100, and incubated at 4°C with LC3B primary rabbit antibody (1:200, #3868, CST, Danvers, MA, USA) overnight. Next, cells were probed with Alexa 594-conjugated goat anti-rabbit secondary antibody (1:200, #8889, CST) at 37°C for 1 h. The images were taken captured using a fluorescence microscope (Carl Zeiss Meditec AG).…”