# Hong-Bai-Lan-Shen Extract Alleviates the CoCl2-Induced Apoptosis in H9C2 Cells by Regulating the AMPK Pathway

## TL;DR

This study shows that Hong-bai-lan-shen extract protects heart cells from hypoxia damage by activating the AMPK pathway, suggesting it could be a new treatment for heart disease.

## Contribution

The study identifies the AMPK pathway as a novel mechanism through which Hong-bai-lan-shen extract protects myocardial cells from hypoxia-induced injury.

## Key findings

- HBLS extract increased cell viability and reduced ROS production in hypoxia-damaged H9C2 cells.
- HBLS extract upregulated AMPK, PI3K, and AKT, and reduced apoptosis markers like caspase-3 and Bax.
- HBLS extract stabilized mitochondrial membrane potential and increased Bcl-2 expression.

## Abstract

In recent years, there has been an increasing number of animal heart diseases caused by various reasons. Heart diseases caused by hypoxia have seriously affected the quality of life of animals. Traditional Chinese medicines such as Rhodiola rosea have shown good efficacy in the treatment of ischemic heart diseases. In this study, based on the principles of traditional Chinese medicine, a scientific formula was developed. Guided by the results of metabolomics, the protective mechanism of Hong-bai-lan-shen extract on H9C2 myocardial cells was investigated in depth. This comprehensive research method is expected to provide a clearer and more reliable theoretical basis for the research and development of new drugs. At the same time, it also helps to gain a deeper understanding of the mechanism of action of traditional Chinese medicine in animal heart health.

This study aims to explore the protective effects of Hong-bai-lan-shen (HBLS) extract, a traditional Chinese medicine compound, on myocardial injury based on metabolomics. H9C2 cells were cultured with HBLS extract for 12 h, and then the cells were cultured in a CoCl2-containing medium, a model simulating the ischemic-hypoxic damage in myocardial cells, for an additional 12 h. The cell viability, cytotoxicity, intracellular metabolite and reactive oxygen species (ROS), mitochondrial membrane potential, apoptosis, and adenosine monophosphate-activated protein kinase (AMPK) signal pathway were determined. The results showed that HBLS extract significantly increased cell viability, stabilized cell morphology, reduced lactate dehydrogenase (LDH) release and ROS production, blocked cysteine-aspartic acid protease 3 (caspase-3) and bcl-2-associated X protein (Bax) expression and decreased apoptotic cell numbers. Meanwhile, HBLS increased membrane potential and the expression of B-cell lymphoma-2 (Bcl-2). Additionally, HBLS extract upregulated the expression of AMPK, PI3K, and protein kinase B (AKT) (p < 0.05, p < 0.01). These findings suggest that HBLS extract has a protective effect on myocardial cells by regulating the AMPK signal pathway and may be a promising therapeutic candidate for ischemic heart disease.