Anisomycin prevents OGD-induced necroptosis by regulating the E3 ligase CHIP

Mi-Bo Tang,Shao-Hua Li,Chang-He Shi,Yuan Cheng,Shuo Zhang,Jonathan C Schisler,Yu-Sheng Li,Yu-Ming Xu,Hai-Yang Luo,Cheng-Yuan Mao,Zheng-Wei Hu

doi:10.1038/s41598-018-24414-y

Abstract

Necroptosis is an essential pathophysiological process in cerebral ischemia-related diseases. Therefore, targeting necroptosis may prevent cell death and provide a much-needed therapy. Ansiomycin is an inhibitor of protein synthesis which can also activate c-Jun N-terminal kinases. The present study demonstrated that anisomycin attenuated necroptosis by upregulating CHIP (carboxyl terminus of Hsc70-interacting protein) leading to the reduced levels of receptor-interacting protein kinase 1 (RIPK1) and receptor-interacting protein kinase 3 (RIPK3) proteins in two in vitro models of cerebral ischemia. Further exploration in this research revealed that losing neither the co-chaperone nor the ubiquitin E3 ligase function of CHIP could abolish its ability to reduce necroptosis. Collectively, this study identifies a novel means of preventing necroptosis in two in vitro models of cerebral ischemia injury through activating the expression of CHIP, and it may provide a potential target for the further study of the disease.

Highlights

Necroptosis is a form of caspase-independent cell death, pathologically characterized by a gain in cell volume, swelling of organelles, plasma membrane rupture, and subsequent loss of intracellular contents[1,2,3,4]
Immunoblot analysis demonstrated that Oxygen-glucose deprivation (OGD) caused an increase in receptor-interacting protein kinase 1 (RIPK1), receptor-interacting protein kinase 3 (RIPK3), p-RIPK3 and p-mixed lineage kinase domain-like (MLKL) (Fig. 1A,B)
This increase in RIPK1, RIPK3, p-RIPK3 and p-MLKL were abrogated by pretreatment with Nec-1 (Fig. 1A,B), suggesting that OGD challenging in a neuronal cell line activates the necroptosis

Summary

Introduction

Necroptosis is a form of caspase-independent cell death, pathologically characterized by a gain in cell volume, swelling of organelles, plasma membrane rupture, and subsequent loss of intracellular contents[1,2,3,4]. Oxygen-glucose deprivation (OGD), an in vitro model of cerebral ischemia, can induce the death receptor (DR)-dependent component of necroptotic cell death in cultured neurons[13,14,15], concomitant with the increase in RIPK3/RIPK1 mRNA and protein levels[16]. Similar phenomenons have been detected in an in vivo model of transient global cerebral ischemia, in the CA1 area[13] These lines of evidence point to inhibiting necroptosis as a novel therapeutic strategy for cerebral ischemic injury, which represents the second leading cause of death as well as the leading cause of premature death and disability. Whether CHIP regulated RIPK1 and RIPK3 degradation occurred in a co-chaperone and/or ubiquitin E3 ligase-dependent manner was assessed by using domain point mutants that disrupt the specific functions of CHIP in cellular model of OGD. The results of this study suggest that CHIP is a candidate therapeutic target for the treatment of necroptosis of ischemic stroke

Methods

Results

Conclusion