Tert-Butylhydroquinone Prevents Oxidative Stress-Mediated Apoptosis and Extracellular Matrix Degradation in Rat Chondrocytes

Wei Lu,Li Cui,Haisheng Huang,Valeria Sülsen,Qisong He,Bo Yang,Lu Zheng

doi:10.1155/2021/1905995

Wei Lu, Li Cui + Show 5 more

Open Access

https://doi.org/10.1155/2021/1905995

Copy DOI

Abstract

Oxidative stress-induced chondrocyte apoptosis and degradation of the extracellular matrix (ECM) play an important role in the progression of osteoarthritis (OA). In addition, tert-butylhydroquinone (TBHQ) is an activator of the nuclear factor erythroid derived-2-related factor 2 (Nrf2). The present study aimed to determine the effectiveness of TBHQ in preventing the apoptosis of chondrocytes and degradation of the extracellular matrix, induced by oxidative stress, in vitro. Therefore, rat chondrocytes were exposed to 20 μM tert-butyl hydroperoxide (TBHP) for 24 h to establish an oxidative damage model, in vitro. Thereafter, cell viability was evaluated using the Cell Counting Kit-8 assay. Moreover, the level of ROS was determined through 2′,7′-dichlorofluorescein diacetate staining. The mitochondrial membrane potential of chondrocytes was also measured using JC-1. Furthermore, cell apoptosis was assessed through Annexin V-fluorescein isothiocyanate/propidium iodide staining. The study also performed Western blotting and qPCR to evaluate the expression of extracellular matrix components, matrix catabolic enzymes, and changes in signalling pathways. The results showed that 2.5 and 5 μM of TBHQ reduced the TBHP-induced generation of excessive ROS and improved cell viability. Additionally, 2.5 and 5 μM of TBHQ prevented mitochondrial damage and apoptosis in rat chondrocytes. Treatment with TBHQ also increased the mRNA and protein expression levels of aggrecan and collagen II. However, TBHQ reduced the mRNA and protein expression levels of matrix metalloproteinase 3 (MMP3) and matrix metalloproteinase 13 (MMP13) in rat chondrocytes. In addition, treatment with TBHQ enhanced the protein expression levels of Nrf2, NADPH quinone oxidoreductase 1 (NQO-1), and hemeoxygenase-1 (HO-1) in rat chondrocytes. The current study showed that TBHQ was not only effective in protecting against TBHP-induced oxidative stress but also inhibited the apoptosis of rat chondrocytes and degradation of the ECM by activating the Nrf2 pathway. The results therefore suggest that TBHQ holds potential for use in the treatment of OA.

Highlights

Osteoarthritis (OA) is the most common form of arthritis that severely affects the patients’ quality of life and causes a substantial clinical and socioeconomic burden
Cell apoptosis was assessed through Annexin V-fluorescein isothiocyanate/propidium iodide staining. e study performed Western blotting and qPCR to evaluate the expression of extracellular matrix components, matrix catabolic enzymes, and changes in signalling pathways. e results showed that 2.5 and 5 μM of TBHQ reduced the tert-butyl hydroperoxide (TBHP)-induced generation of excessive ROS and improved cell viability
Treatment with TBHQ enhanced the protein expression levels of nuclear factor erythroid derived-2-related factor 2 (Nrf2), NADPH quinone oxidoreductase 1 (NQO-1), and hemeoxygenase-1 (HO-1) in rat chondrocytes. e current study showed that TBHQ was effective in protecting against TBHP-induced oxidative stress and inhibited the apoptosis of rat chondrocytes and degradation of the extracellular matrix (ECM) by activating the Nrf2 pathway. e results suggest that TBHQ holds potential for use in the treatment of OA

Summary

Introduction

Osteoarthritis (OA) is the most common form of arthritis that severely affects the patients’ quality of life and causes a substantial clinical and socioeconomic burden. Erefore, inhibiting the oxidative stress-induced apoptosis of chondrocytes and ECM degradation may be an effective means of preventing cartilage degeneration and progression of OA. TBHQ was proven to have strong antioxidant effects through the activation of the Nrf pathway, in in vitro or in vivo studies of some diseases such as alcoholic cardiomyopathy, acute hepatic injury, and neurodegenerative disease [15,16,17,18]. It is still unknown whether TBHQ can inhibit the apoptosis of chondrocytes and ECM degradation, induced by oxidative stress. Erefore, the present study attempted to investigate the ability of TBHQ to inhibit the apoptosis of chondrocytes and ECM degradation, induced by oxidative stress. e possible underlying mechanisms were assessed

Materials and Methods

Results

Discussion