Section snippets
INTRODUCTION
RA is a systemic autoimmune disease primarily affecting the joints. The risk factors for RA include both modifiable lifestyle-related factors and non-modifiable traits such as heredity and gender, with a higher incidence among females (Radu and Bungau, 2021). In RA, joints exhibit synovial inflammation, destruction of articular cartilage, and erosion of bones, ultimately leading to joint deformity and disability (Sparks, 2019). Articular cartilage consists solely of chondrocytes, which play a
Cell culture
Human C28/I2 cells (Millipore, USA) were cultured in Dulbecco’s modified eagle medium (DMEM, Hyclone, United States) containing 10% fetal bovine serum (FBS, Wisent, Canada) and 1% penicillin-streptomycin (Beyotime, China) in a sterile wet atmosphere containing 5% CO2 at 37 °C. The culture medium was changed every 2-3 days. Toluidine blue and type II collagen staining were used to identify chondrocytes.
MTT assay
Cell viability was evaluated by MTT. Human C28/I2 cells were seeded in a 96-well plate
NS8593 inhibits chondrocyte ferroptosis induced by Erastin
In order to verify the effect of NS8593 on ferroptosis of chondrocytes, we established an in vitro model of ferroptosis in human chondrocytes ( C28/I2 cells) using the ferroptosis inducer Erastin. Our results showed that Erastin reduced cell viability and increased cytotoxicity, while NS8593 inhibited Erastin-induced chondrocyte death (Fig. 1b-e). Considering that NS8593 is not only an inhibitor of TRPM7, but also an inhibitor of SK channels. Therefore, we used Apamin, a specific inhibitor of
DISCUSSION
In this study, we investigated the role of NS8593, a TRPM7 channel inhibitor, on articular cartilage damage in CFA-induced AA rat and its effect on Erastin-induced ferroptosis in human chondrocytes, as well as the potential association and mechanism of action of TRPM7 and HO-1. To begin, we established a model of Erastin-induced ferroptosis in human C28/I2 cells and observed a significant reduction in chondrocyte viability upon treatment with 2 μM Erastin. However, this detrimental effect was
CONCLUSION
In conclusion, we found for the first time that NS8593 inhibits ferroptosis in articular chondrocytes and reduces cartilage damage in AA rats, and we also found that the TRPM7/HO-1 signaling pathway is a key regulatory target for NS8593 to inhibit Erastin-induced chondrocyte ferroptosis. Targeting TRPM7/HO-1 may be an effective treatment for controlling articular cartilage injury in RA, and suggest that NS8593 may be a potential new drug for the treatment of RA.
Ethics statement
The animal study was reviewed and approved by the Ethical Regulations for the Care and Use of Laboratory Animals of Anhui Medical University（LLSC20190062）.
Author contributions
Renpeng Zhou, Wei Hu, Wenjuan Hao and Rendi Zhu conceived and designed this research. Wenjuan Hao, Rendi Zhu, Yong Chen, Hailin Zhang, Shufang Li, Fuli Zhou performed the in vivo and in vitro experiments. The data analysis was done by Wenjuan Hao, Rendi Zhu and Hailin Zhang. Renpeng Zhou, Wenjuan Hao and Rendi Zhu participated in writing the manuscript. All authors have approved the final version of the manuscript.
Funding
This work was supported by grants from the National Natural Science Foundation of China (No.82272450, No.82371575), and the Outstanding Youth Project of Anhui Provincial Natural Science Foundation (2308085Y44).
CRediT authorship contribution statement
Hailin Zhang: Writing – review & editing, Software, Resources, Investigation, Data curation. Yong Chen: Writing – review & editing, Validation, Methodology. Shufang Li: Supervision, Project administration, Methodology. Fuli Zhou: Validation, Software. Wei Hu: Writing – review & editing, Supervision, Project administration, Conceptualization. Renpeng Zhou: Writing – review & editing, Supervision, Funding acquisition, Conceptualization. Wenjuan Hao: Writing – review & editing, Writing – original
Declaration of competing interest
The authors declare no competing interests.
References (41)
Targeting regulated chondrocyte death in osteoarthritis therapy
Biochem Pharmacol
(2023)
TRPM7 senses oxidative stress to release Zn(2+) from unique intracellular vesicles
Proc Natl Acad Sci U S A
(2017)
Prolactin promotes cartilage survival and attenuates inflammation in inflammatory arthritis
J Clin Invest
(2013)
Natural and synthetic modulators of SK (K(ca)2) potassium channels inhibit magnesium-dependent activity of the kinase-coupled cation channel TRPM7
Br J Pharmacol
(2012)
Targeted apoptosis of macrophages and osteoclasts in arthritic joints is effective against advanced inflammatory arthritis
Nat Commun
(2021)
Inhibition of small-conductance Ca2+-activated K+ channels terminates and protects against atrial fibrillation
Circ Arrhythm Electrophysiol
(2010)
TRPM7-mediated Ca2+ signals confer fibrogenesis in human atrial fibrillation
Circ Res
(2010)
Ferroptosis as a target for protection against cardiomyopathy
Proc Natl Acad Sci U S A
(2019)
TRPM7 Kinase Controls Calcium Responses in Arterial Thrombosis and Stroke in Mice
Arterioscler Thromb Vasc Biol
(2018)
View full text
© 2024 Elsevier Ltd. All rights are reserved, including those for text and data mining, AI training, and similar technologies.