Tongyang Huoxue Decoction (TYHX) Ameliorating Hypoxia/Reoxygenation-Induced Disequilibrium of Calcium Homeostasis and Redox Imbalance via Regulating Mitochondrial Quality Control in Sinoatrial Node Cells.

Xing Chang,Shunyu Yao,Yanli Wang,Ruxiu Liu,Jinfeng Liu,Qiaomin Wu,Jin Wang

doi:10.1155/2021/3154501

Abstract

Sick sinus syndrome (SSS) is a disease with bradycardia or arrhythmia. The pathological mechanism of SSS is mainly due to the abnormal conduction function of the sinoatrial node (SAN) caused by interstitial lesions or fibrosis of the SAN or surrounding tissues, SAN pacing dysfunction, and SAN impulse conduction accompanied by SAN fibrosis. Tongyang Huoxue Decoction (TYHX) is widely used in SSS treatment and amelioration of SAN fibrosis. It has a variety of active ingredients to regulate the redox balance and mitochondrial quality control. This study mainly discusses the mechanism of TYHX in ameliorating calcium homeostasis disorder and redox imbalance of sinoatrial node cells (SANCs) and clarifies the protective mechanism of TYHX on the activity of SANCs. The activity of SANCs was determined by CCK-8 and the TUNEL method. The levels of apoptosis, ROS, and calcium release were analyzed by flow cytometry and immunofluorescence. The mRNA and protein levels of calcium channel regulatory molecules and mitochondrial quality control-related molecules were detected by real-time quantitative PCR and Western Blot. The level of calcium release was detected by laser confocal. It was found that after H/R treatment, the viability of SANCs decreased significantly, the levels of apoptosis and ROS increased, and the cells showed calcium overload, redox imbalance, and mitochondrial dysfunction. After treatment with TYHX, the cell survival level was improved, calcium overload and oxidative stress were inhibited, and mitochondrial energy metabolism and mitochondrial function were restored. However, after the SANCs were treated with siRNA (si-β-tubulin), the regulation of TYHX on calcium homeostasis and redox balance was counteracted. These results suggest that β-tubulin interacts with the regulation of mitochondrial function and calcium release. TYHX may regulate mitochondrial quality control, maintain calcium homeostasis and redox balance, and protect SANCs through β-tubulin. The regulation mechanism of TYHX on mitochondrial quality control may also become a new target for SSS treatment.

Highlights

Sick sinus syndrome (SSS), known as sinoatrial node dysfunction, is a syndrome of arrhythmias caused by the pathological changes of sinoatrial node (SAN) and its adjacent tissues
We first verified the effect of Tongyang Huoxue Decoction (TYHX) on activity of sinoatrial node cells (SANCs), and it was found that TYHX can further improve cell viability through CCK-8 detection (Figure 1(a)), while effectively inhibiting cell apoptosis (Figures 1(e) and 1(f))
In order to further verify the role of skeleton protein (β-tubulin) in SANC damage induced by hypoxia/reoxygenation (H/R), we detected the levels of βtubulin by PCR/Western Blot and immunofluorescence

Summary

Introduction

Sick sinus syndrome (SSS), known as sinoatrial node dysfunction, is a syndrome of arrhythmias caused by the pathological changes of sinoatrial node (SAN) and its adjacent tissues. It is one of the most refractory major cardiovascular diseases in clinic [1]. Studies have found that calcium homeostasis and oxidative stress can affect SAN function directly [4]. Mitochondrial quality control (MQC) is closely related to calcium homeostasis and redox balance [5, 6], which directly affect the survival level of sinoatrial node cells (SANCs). Its normal structure and function can meet the energy requirements of heart beating and ejection function, maintaining the homeostasis of intracellular environment, and regulating cell growth [8, 9]

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