Eleutheroside E inhibits doxorubicin-induced inflammation and apoptosis in rat cardiomyocytes by modulating activation of NF-κB pathway

Zheng Wu,Yuhua Sun,Wenzheng Li,Shujuan Cheng,Kun Fu

doi:10.4314/tjpr.v16i3.4

Abstract

Purpose : To identify the effects of eleutheroside E (EE) on apoptosis and inflammation induced by doxorubicin (DOX) in H9c2 cells and to investigate the underlying mechanisms. Methods : The effect of EE on H9c2 cell viability was determined using Cell Counting Kit-8 (CCK8). EE effect on DOX-induced apoptosis and inflammation in H9c2 cells was studied by comparison between cells treated with DOX alone and DOX+EE; the relationship between EE effects and NF-κB signaling pathway was evaluated by the addition of NF-κB inhibitor PDTC. Cell apoptosis was examined by flow cytometry while IL-1β, IL-6, and TNF-α levels were determined by ELISA. The phosphorylation level of NF-κB p65 was measured by Western blot. Results : Compared with control group, cell viability was notably elevated after treatment with 50-100 μM EE for 48 or 72 h. DOX induced higher rates of cell apoptosis in H9c2 cells (29.5 ± 3.56 %) compared with control group (6.39 ± 0.67 %); however, with EE pretreatment (50 and 80 μM), apoptosis rate decreased to 16.8 ± 2.16 and 13.54 ± 2.08 %, respectively, which are significantly lower than that of DOX group; furthermore, the levels of IL-1β, IL-6, and TNF-α also reduced. In addition, DOX-induced phosphorylation of NF-κB p65 was suppressed by EE pretreatment (10, 50 and 80 μM) to 11.51 ± 1.25, 40.2 ± 5.17 and 52.97 ± 6.74 %, respectively Conclusion : The results suggest that EE treatment reduced DOX-induced apoptosis and inflammation by interacting with NF-κB signaling pathway. This finding sheds some light on probable new strategies on the application of DOX for cancer treatment. Keywords : Eleutheroside E, Doxorubicin, Inflammation, Apoptosis, Cardiomyocytes, NF-κB

Highlights

Doxorubicin (DOX), one of the most commonly used anti-cancer drugs, possesses therapeutic effects for a variety of cancers, including leukemia, lymphoma, soft tissue sarcomas and solid tumors [1,2,3,4]
Eleutheroside E (EE) inhibits H9c2 cell apoptosis induced by DOX
We looked into whether or not EE influenced H9c2 cell apoptosis induced by DOX

Summary

INTRODUCTION

Doxorubicin (DOX), one of the most commonly used anti-cancer drugs, possesses therapeutic effects for a variety of cancers, including leukemia, lymphoma, soft tissue sarcomas and solid tumors [1,2,3,4]. 3hydroxy-3-methylglutaryl coenzyme A (reductase inhibitor, which has anti-inflammatory and antioxidative effects) has been used to lessen the cardiotoxicity of DOX in mice [11] Both suppressed cardiac cytokine activation and lipid peroxidation have been shown to play critical roles in improving left ventricular (LV). To examine the effects of EE on DOX (SigmaAldrich, St. Louis, MO, USA) induced injury, H9c2 cells were divided into five groups: control, DOX(1μM), EE (10 μM), EE (50 μM) and EE (80 μM) groups. Cell Counting Kit-8 (CCK8) (Sigma-Aldrich, St. Louis, MO, USA) solution (10 μL) was added to each well at 1/10 dilution, followed by incubation of 2 hours. Proteins in cytoplasm and nuclei from H9c2 cells were isolated by the NE-PER Nuclear and Cytoplasmic Extraction Kit according to the manufacturer’s protocol (Thermo Fisher Scientific, Waltham, MA, USA). A value of p < 0.05 was considered statistically significant

RESULTS

DISCUSSION

CONCLUSION

Conflict of Interest