3,3'‑Diindolylmethane attenuates cardiomyocyte hypoxia by modulating autophagy in H9c2 cells.

Abstract

Autophagy is activated in the ischemic heart and is a process that is essential for survival, differentiation, development and homeostasis. 3,3'‑Diindolylmethane (DIM) is a natural product of the acid‑catalyzed condensation of indole‑3‑carbinol in cruciferous vegetables. Numerous studies have suggested that DIM has various pharmacological effects, including antioxidant, antitumor, anti‑angiogenic and anti‑apoptotic properties. However, the function of DIM on hypoxia‑induced cardiac injury remains to be elucidated. In the present study, H9c2 cells were pretreated with DIM (1, 5 and 10µM) for 12h and exposed to hypoxia for 12h. It was demonstrated that DIM markedly attenuated the increased transcription of interleukin (IL)‑1β, IL‑6 and tumor necrosis factor‑α induced by hypoxia. In addition, the transcription of autophagy associated genes increased in the DIM pretreated group, compared with the hypoxia group. DIM additionally attenuated the increased apoptosis, as determined by terminal deoxynucleotidyl transferase dUTP nick end labeling staining, and regulated the relative protein expression level of B cell lymphoma (Bcl) 2 associated X protein, Bcl‑xL and cleaved caspase 3. Furthermore, the phosphorylation of the 5'AMP‑activated protein kinase a (AMPKa) was activated and the phosphorylation of c‑Jun N‑terminal kinase (JNK) was inhibited. The effect of DIM on hypoxia‑induced apoptosis was abolished following pretreatment with JNK activator (anisomycin, 40ng/ml). The effect of DIM on autophagy was reversed following pretreatment with AMPKa inhibitor (CpC, 20µM) following stimulation with hypoxia. The results demonstrated that DIM prevented hypoxia‑induced inflammation and apoptosis and activated cardiomyocyte autophagy, which may be mediated by activation of AMPKa and inhibition of JNK pathways.