1. Academic Validation
  2. Neferine suppresses autophagy-induced inflammation, oxidative stress and adipocyte differentiation in Graves' orbitopathy

Neferine suppresses autophagy-induced inflammation, oxidative stress and adipocyte differentiation in Graves' orbitopathy

  • J Cell Mol Med. 2021 Feb;25(4):1949-1957. doi: 10.1111/jcmm.15931.
Hong Li 1 Long Gao 1 Jie Min 1 Yucheng Yang 1 Ren Zhang 1
Affiliations

Affiliation

  • 1 Department of Endocrinology, Shanghai University of Traditional Chinese Medicine Longhua affiliated Hospital, Shanghai, China.
Abstract

Previous studies in Graves' orbitopathy (GO) patient-derived fibroblasts showed that inhibition of Autophagy suppresses adipogenic differentiation. Autophagy activation is associated with inflammation, production of Reactive Oxygen Species and fibrosis. Neferine is an alkaloid extracted from Nelumbo nucifera, which induces Nrf2 expression and inhibits Autophagy. Here, we have elucidated the role of neferine on interleukin (IL)-13-induced Autophagy using patient-derived orbital fibroblasts as an in vitro model of GO. GO patient-derived orbital fibroblasts were isolated and cultured to generate an in vitro model of GO. Autophagy was determined by Western blot detection of the markers such as Beclin-1, Atg-5 and LC3 and by immunofluorescence detection of autophagosome formation. Analysis of differentiation towards an adipogenic lineage was performed by Oil red O staining. The expression of inflammatory factors was detected by ELISA and semiquantitative RT-PCR. Neferine inhibited Autophagy in GO orbital fibroblasts, as indicated by the suppression of IL-13-induced autophagosome formation, overexpression of Autophagy markers, increased LC3-II/LC3-I levels and finally down-regulation of p62. Neferine suppressed IL-13-induced inflammation, ROS generation, fibrosis and adipogenic differentiation in GO patient-derived orbital fibroblasts. The anti-inflammatory, antioxidant and antiadipogenic effects of neferine were accompanied by the up-regulation of Nrf2. These results indicated that orbital tissue remodelling and inflammation in GO may be mediated by Autophagy, and neferine suppressed autophagy-related inflammation and adipogenesis through a mechanism involving Nrf2.

Keywords

Graves’ orbitopathy; Neferine; adipocyte differentiation; autophagy; inflammation; oxidative stress.

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