Ethanol extracts of Rhaponticum uniflorum (L.) DC flowers attenuate doxorubicin-induced cardiotoxicity via alleviating apoptosis and regulating mitochondrial dynamics in H9c2 cells

Boqin Hu; Dong Zhen; Meirong Bai; Tianqi Xuan; Yu Wang; Mingjie Liu; Lijun Yu; Dongsong Bai; Danni Fu; Chengxi Wei

doi:10.1016/j.jep.2021.114936

Ethanol extracts of Rhaponticum uniflorum (L.) DC flowers attenuate doxorubicin-induced cardiotoxicity via alleviating apoptosis and regulating mitochondrial dynamics in H9c2 cells

J Ethnopharmacol. 2022 Apr 24:288:114936. doi: 10.1016/j.jep.2021.114936. Epub 2022 Jan 7.

Authors

Boqin Hu¹, Dong Zhen², Meirong Bai³, Tianqi Xuan⁴, Yu Wang⁵, Mingjie Liu⁶, Lijun Yu⁷, Dongsong Bai⁸, Danni Fu⁹, Chengxi Wei¹⁰

Affiliations

¹ Institute of Pharmaceutical Chemistry and Pharmacology, Inner Mongolia Minzu University, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China; Inner Mongolia Key Laboratory of Mongolian Medicine Pharmacology for Cardio-Cerebral Vascular System, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: 839061084@qq.com.
² Institute of Pharmaceutical Chemistry and Pharmacology, Inner Mongolia Minzu University, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China; Inner Mongolia Key Laboratory of Mongolian Medicine Pharmacology for Cardio-Cerebral Vascular System, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: dongzhen@imun.edu.cn.
³ Key Laboratory of Mongolian Medicine Research and Development Engineering, Ministry of Education, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: baimeirong@126.com.
⁴ Institute of Pharmaceutical Chemistry and Pharmacology, Inner Mongolia Minzu University, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China; Inner Mongolia Key Laboratory of Mongolian Medicine Pharmacology for Cardio-Cerebral Vascular System, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: 1543505206@qq.com.
⁵ Institute of Pharmaceutical Chemistry and Pharmacology, Inner Mongolia Minzu University, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China; Inner Mongolia Key Laboratory of Mongolian Medicine Pharmacology for Cardio-Cerebral Vascular System, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: shen348@126.com.
⁶ Institute of Pharmaceutical Chemistry and Pharmacology, Inner Mongolia Minzu University, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China; Inner Mongolia Key Laboratory of Mongolian Medicine Pharmacology for Cardio-Cerebral Vascular System, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: lmjlwz@163.com.
⁷ Institute of Pharmaceutical Chemistry and Pharmacology, Inner Mongolia Minzu University, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China; Inner Mongolia Key Laboratory of Mongolian Medicine Pharmacology for Cardio-Cerebral Vascular System, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: tl_ylj@163.com.
⁸ Institute of Pharmaceutical Chemistry and Pharmacology, Inner Mongolia Minzu University, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China; Inner Mongolia Key Laboratory of Mongolian Medicine Pharmacology for Cardio-Cerebral Vascular System, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: 1605551902@qq.com.
⁹ Institute of Pharmaceutical Chemistry and Pharmacology, Inner Mongolia Minzu University, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China; Inner Mongolia Key Laboratory of Mongolian Medicine Pharmacology for Cardio-Cerebral Vascular System, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: biops2016@163.com.
¹⁰ Institute of Pharmaceutical Chemistry and Pharmacology, Inner Mongolia Minzu University, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China; Inner Mongolia Key Laboratory of Mongolian Medicine Pharmacology for Cardio-Cerebral Vascular System, Tongliao, 028000, Inner Mongolia Autonomous Region, PR China. Electronic address: weichengxi1224@163.com.

PMID: 35007682
DOI: 10.1016/j.jep.2021.114936

Abstract

Ethnopharmacological relevance: Loulu flowers (LLF) is the inflorescence of Rhaponticum uniflorum (L.) DC. (R. uniflorum), a member of the Compositae family. This plant possesses heat-clearing properties, detoxification effects, and is therefore frequently used for the treatment of cardiovascular diseases.

Aim of this study: This study aimed to investigate the cardioprotective effects of ethanol extracts of LLF against doxorubicin (DOX)-induced cardiotoxicity and explore the associated mechanisms.

Material and methods: Ethanol extracts of LLF were prepared and analyzed by LC-ESI-MS/MS. DOX-treated H9c2 cells and DOX-treated zebrafish models were used to explore the cardioprotective effect of ethanol extracts on myocardial function. The effects of LLF on DOX-induced cytotoxicity in H9c2 cells were investigated by MTT assay. Reactive Oxygen Species (ROS) levels, mitochondrial membrane potential (MMP), and nuclear translocation of NF-κB p65 were examined using fluorescent probes. The expression level of Bax, Bcl-2, PARP, caspase-3, cleaved-caspase3, caspase9, IκBα, p-IκBα, IKK, p-IKK, p65, p-p65, OPA1, Mfn1, MFF and Fis 1 and GAPDH was determined by western blotting.

Results: Twenty-five compounds were detected in ethanol extracts of LLF, include Nicotinamide, Coumarin, Parthenolide, and Ligustilide. Pre-treatment with LLF attenuated the DOX-induced decrease in viability and ROS production in H9c2 cells. Moreover, LLF treatment maintained the mitochondrial membrane integrity and suppressed apoptosis by upregulating expression level of Bcl-2 and downregulating the expression level of Bax, cleaved-caspase-3, cleaved-caspase-9 and cleaved-PARP. In addition, LLF significantly inhibited the DOX-induced activation of NF-κB signaling. Cells treated with DOX showed aberrant expression of mitochondrial dynamics related proteins, and these effects were alleviated by LLF pre-treatment. In conclusion, these results show that LLF can alleviate DOX-induced cardiotoxicity by blocking NF-κB signaling and re-balancing mitochondrial dynamics.

Conclusion: Ethanol extracts of LLF is a potential treatment option to against DOX-induced cardiotoxicity.

Keywords: Doxorubicin; Loulu flowers; Mitochondrial dynamics; Myocardial protective effect.

MeSH terms

Animals
Antibiotics, Antineoplastic / toxicity
Apoptosis / drug effects
Cardiotonic Agents / isolation & purification
Cardiotonic Agents / pharmacology
Cardiotoxicity / etiology
Cardiotoxicity / prevention & control*
Cell Line
Doxorubicin / toxicity*
Ethanol / chemistry
Female
Leuzea / chemistry*
Male
Mitochondrial Dynamics / drug effects
Myocytes, Cardiac / drug effects
Myocytes, Cardiac / pathology
Plant Extracts / pharmacology*
Rats
Tandem Mass Spectrometry
Zebrafish

Substances

Antibiotics, Antineoplastic
Cardiotonic Agents
Plant Extracts
Ethanol
Doxorubicin