Nrf2 drives oxidative stress-induced autophagy in nucleus pulposus cells via a Keap1/Nrf2/p62 feedback loop to protect intervertebral disc from degeneration

Zehan Tang,Bo Hu,Xingda Zhang,Jianxi Wang,Huajiang Chen,Fazhi Zang

doi:10.1038/s41419-019-1701-3

Abstract

Intervertebral disc (IVD) degeneration is known to aggravate with age and oxidative stress is implicated in the pathogenesis of many age-related diseases. Nuclear factor (erythroid-derived-2)-like 2 (Nrf2) can confer adaptive protection against oxidative and proteotoxic stress in cells. In this study, we assessed whether Nrf2 can protect against oxidative stress in nucleus pulposus (NP) cells. In addition, we investigated Nrf2 expression in NP tissue samples from patients with different degrees of IVD degeneration and a mouse model of aging and IVD degeneration and the influence of H2O2-induced oxidative stress on autophagic pathways in NP cells. Autophagy was assessed by measuring levels of autophagy-related protein (ATG) family members and the autophagic markers, p62 and LC3. We found that expression of Nrf2 progressively decreased in human NP tissue samples of patients with increasing degrees of IVD degeneration. Nrf2 deficiency leads to the degeneration of IVDs during aging. Nrf2 knockout also aggravates IVD degeneration and reduces autophagic gene expression in an induced mouse model of IVD degeneration. The detrimental effects of H2O2-induced oxidative stress were increased in autophagy-deficient cells via reduced expression of Atg7 and the Keap1–Nrf2–p62 autophagy pathway. Taken together, these results suggest that excessive oxidative stress causes the upregulation of autophagy, and autophagy acts as an antioxidant feedback response activated by a Keap1-Nrf2-p62 feedback loop in IVD degeneration.

Highlights

Intervertebral discs (IVDs) are subject to degeneration as they age, owing to changes in the abundance and structure of macromolecules[1,2]
nucleus pulposus (NP) tissue samples from patients with different degrees of IVD degeneration were divided into four groups according to a previously described classification system[27]: non-degenerated, mildly degenerated, moderately degenerated, and severely degenerated (Fig. 1)
Atg[7] siRNA reduced while Keap[1] siRNA increased light chain 3 (LC3) expression (Fig. 5f) which was contrary to p62 (Fig. 5h) in NP cells under H2O2induced oxidative stress. These results indicate that H2O2-induced oxidative stress was increased in autophagy-deficient cells via an autophagic pathway modulated by Atg[7] and the Kelch-like ECH-associated protein 1 (Keap1)–Nuclear factor (erythroid-derived-2)-like 2 (Nrf2)–p62

Summary

Introduction

Intervertebral discs (IVDs) are subject to degeneration as they age, owing to changes in the abundance and structure of macromolecules[1,2]. The center of the disc contains the nucleus pulposus (NP), which is a highly hydrated gelatinous aggrecan-rich core (Urban and Roberts[3]). The NP is surrounded by collagen I-rich fibrous cartilage known as the annulus fibrosus (AF). The NP becomes more fibrous and less gelatinous and the boundary between the NP and AF. Autophagy is an evolutionarily conserved homeostatic process involved in quality control and the recycling of cell components[6]. The autophagic process is initiated by a complex involving unc-51-like kinase 1 (ULK1) and members of the autophagy-related protein (ATG) family[7]. The ULK1 complex translocates to Official journal of the Cell Death Differentiation Association

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Conclusion