Mechanistic Insights into Salidroside's Mitochondrial Protection via AMPK/Sirt1/HIF-1&#945; Pathway in Hypoxic HT22 Cells

Xiaobo Wang; Yating Zhang; Ya Hou; Hong Jiang; Tingting Kuang; Rui Li; Xianli Meng

doi:10.3791/66923

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Research Article

Mechanistic Insights into Salidroside's Mitochondrial Protection via AMPK/Sirt1/HIF-1α Pathway in Hypoxic HT22 Cells

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DOI:

10.3791/66923

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April 25th, 2025

Xiaobo Wang¹^,²^,³ , Yating Zhang¹ , Ya Hou¹ , Hong Jiang¹ , Tingting Kuang¹ , Rui Li³ , Xianli Meng¹^,²^,³

¹School of Pharmacy/School of Modern Chinese Medicine Industry, Chengdu University of Traditional Chinese Medicine, ²Innovative Institute of Chinese Medicine and Pharmacy/Academy for Interdiscipline, Chengdu University of Traditional Chinese Medicine, ³Research Service Office, Traditional Chinese Medicine Hospital of Meishan

Summary

The present study identified a unique mechanism by which salidroside exerts mitochondrial protective effects on hypoxic HT22 cells, partly through the AMPK/Sirt1/HIF-1α pathway.

Abstract

Salidroside (Sal), an active ingredient of Rhodiola crenulata (Hook. f. et Thoms.) H. Ohba has been found to exert mitochondrial protective effects by improving metabolism and enhancing the energy supply of brain cells under hypoxic conditions. However, its mechanism of action has not been fully clarified. In the present study, high-performance liquid chromatography was first employed to analyze the effects of Sal on nucleotide (ATP, ADP, and AMP) levels. The cellular thermal shift assay (CETSA), a widely used molecular interaction method for validating and quantifying drug target engagement in cells and tissues across different species, was then chosen to confirm the affinity of Sal for AMPK/Sirt1/HIF-1α pathway-related proteins. The results revealed that Sal increased ATP and ADP levels in hypoxic HT22 cells while reducing AMP levels. Moreover, Sal exhibited stable binding to AMPKα, p-AMPKα, Sirt1, and HIF-1α proteins. In conclusion, Sal may exert mitochondrial protective effects by modulating the AMPK/Sirt1/HIF-1α pathway to regulate nucleotide content. This study provides a methodological reference for nucleotide content analysis in cell samples and contributes to the identification and discovery of targets for compounds derived from traditional Chinese medicine.

Introduction

The brain is highly sensitive to oxygen due to its high metabolic demands, limited glycolytic capacity, and dependence on oxidative phosphorylation. As a result, exposure to a low-oxygen environment at high altitudes can readily lead to hypobaric hypoxic brain injury (HHBI)¹^,². Epidemiological studies indicate that when individuals unacclimated to high altitudes ascend rapidly to high-altitude regions, the incidence of acute mountain sickness can reach up to 75%, with a fatality rate of approximately 1% for severe cases. Furthermore, in the absence of medical care, mortality rates for high-altitude cerebral or....