TGFβ1 signaling protects chondrocytes against oxidative stress via FOXO1-autophagy axis

Objective: The forkhead box O1 (FOXO1) transcription factor is a key regulator of autophagy. In chondrocytes, reduced FOXO1 expression with aging causes osteoarthritis due to dysfunction of autophagy, but the mechanisms underlying regulation of FOXO1 expression and the reduction in expression with aging remain unclear. We investigated the mechanism by which transforming growth factor beta 1 (TGF beta 1) signaling regulates the FOXO1-autophagy axis. Methods: Expression of FOXO1 was measured in chondrocytes after TGF beta 1 treatment. Immunohistochemistry was performed to estimate the levels of activin receptor-like kinase 5 (ALK5) and FOXO1 in the knee joints of young, middle-aged and old mice. The effects of the ALK5 inhibitor and SMAD3 or SMAD2 knockdown on FOXO1 expression were evaluated. The role of TGF beta 1 in autophagy after hydrogen peroxide (H2O2) treatment was analyzed. The protective effect of TGF beta 1 against H2O2 treatment was assessed by cell viability assay and TUNEL assay. Results: TGF beta 1 promoted the expression of FOXO1 mRNA and protein. Both ALK5 and FOXO1 expression decreased with aging. ALK5 inhibition and SMAD3 knockdown suppressed induction of FOXO1 expression by TGF beta 1, whereas SMAD2 knockdown increased it. TGF beta 1 promoted the expression of microtubule-associated proteins 1A/1B light chain 3B (LC3)-I protein via the SMAD3-FOXO1 pathway. Furthermore, under H2O2 treatment, TGF beta 1 promoted expression of LC3-II. TGF beta 1 pretreatment suppressed cell death of chondrocytes following H2O2 treatment, but this protective effect was abolished by FOXO1 knockdown. Conclusions: TGF beta 1 protects chondrocytes against oxidative stress via the FOXO1-autophagy axis, and a reduction in ALK5 expression might cause reduced FOXO1 expression with aging. (C) 2021 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

Automated summary

FOXO1, a key regulator of autophagy, is promoted by TGF-β1 to protect chondrocytes against oxidative stress. Reduced ALK5 expression with aging may lead to decreased FOXO1 expression.