RIP3 facilitates necroptosis through CaMKII and AIF after intracerebral hemorrhage in mice

Neurosci Lett. 2021 Apr 1;749:135699. doi: 10.1016/j.neulet.2021.135699.

Yang Xu ¹, Xiaodong Wu ², Wenjie Hu ³, Dijing Yu ⁴, Zhiding Shao ⁵, Weifei Li ⁶, Tingting Huang ⁷, Ji Zhang ⁸, Xiaolong Zhu ⁹, Xueqin Li ¹⁰, Hui Yang ¹¹, Zhaohu Chu ¹², Kun Lv ¹³

Affiliations

1 Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institutes, Wannan Medical College, Wuhu, 241001, Anhui, China; Non-coding RNA Research Center of Wannan Medical College, Wuhu, 241001, Anhui, China; Department of Neurology, The First Affiliated Hospital of Wannan Medical College, Wuhu, 241001, Anhui, China. Electronic address: xuyang@wnmc.edu.cn.
2 Department of Neurology, The Second Affiliated Hospital of Wannan Medical College, Wuhu, 241000, Anhui, China. Electronic address: wuxiaodong131415@163.com.
3 Department of Neurology, The First Affiliated Hospital of Wannan Medical College, Wuhu, 241001, Anhui, China. Electronic address: huwenjie190@163.com.
4 Department of Ophthalmology, Wuhu Eye Hospital, Wuhu, 241000, Anhui, China. Electronic address: ydjydj860128@126.com.
5 Department of Neurology, The Second Affiliated Hospital of Wannan Medical College, Wuhu, 241000, Anhui, China. Electronic address: shaozhiding@126.com.
6 Department of Neurology, The Second Affiliated Hospital of Wannan Medical College, Wuhu, 241000, Anhui, China. Electronic address: lbqlwf@163.com.
7 Department of Neurology, The First Affiliated Hospital of Wannan Medical College, Wuhu, 241001, Anhui, China. Electronic address: 1006551570@qq.com.
8 Department of Neurology, The First Affiliated Hospital of Wannan Medical College, Wuhu, 241001, Anhui, China. Electronic address: 1282295518@qq.com.
9 Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institutes, Wannan Medical College, Wuhu, 241001, Anhui, China; Non-coding RNA Research Center of Wannan Medical College, Wuhu, 241001, Anhui, China. Electronic address: zhuxlon3@126.com.
10 Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institutes, Wannan Medical College, Wuhu, 241001, Anhui, China; Non-coding RNA Research Center of Wannan Medical College, Wuhu, 241001, Anhui, China. Electronic address: lixueqin0727@163.com.
11 Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institutes, Wannan Medical College, Wuhu, 241001, Anhui, China; Non-coding RNA Research Center of Wannan Medical College, Wuhu, 241001, Anhui, China. Electronic address: tjykdxyh0203@163.com.
12 Department of Neurology, The First Affiliated Hospital of Wannan Medical College, Wuhu, 241001, Anhui, China. Electronic address: chuzhaohu878@163.com.
13 Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institutes, Wannan Medical College, Wuhu, 241001, Anhui, China; Non-coding RNA Research Center of Wannan Medical College, Wuhu, 241001, Anhui, China. Electronic address: lvkun315@126.com.

PMID: 33540056 DOI: 10.1016/j.neulet.2021.135699

Abstract

Background: Necroptosis-induced neuronal damage after intracerebral hemorrhage (ICH) has been documented recently. Previous studies have reported that RIP3 and its complex are recognized as central mediators of Necroptosis. In this study, the role of RIP3 in the activation of CaMKII and AIF was investigated.

Methods: We induced ICH in C57BL/6 mice by injecting collagenase IV into the basal ganglia. ICH mice were pretreated with the mPTP inhibitor CsA and the CAMKII inhibitor Kn-93, RIP3 siRNA or RIP3 rAAV. Brain edema and neurobehavior were evaluated. The expression of RIP3, p-MLKL, AIF, and CaMKII proteins was evaluated by western blotting, immunofluorescence (IF) and immunoprecipitation (IP).

Results: Significant increases in RIP3, p-MLKL, CaMKII and AIF expression were observed in ICH mice, and RIP3-AIF colocalized in the nucleus. Overexpression of RIP3 by rAAV upregulated AIF expression in both the cytoplasm and nucleus, while CaMKII expression was increased in the cytoplasm. The interaction of RIP3-AIF and RIP3-CaMKII was detected after ICH injury. These complexes were inhibited by CsA with Kn-93 or RIP3 siRNA pretreatment, which reduced brain edema and neurological deficits.

Conclusions: Our findings revealed that ICH induced necroptotic neuronal death through the RIP3-CaMKII complex and the RIP3-AIF signaling pathway. Moreover, blockade of mPTP opening could suppress the pathogenesis of Necroptosis.

Keywords

Apoptosis-inducing factor (AIF); Ca(2+)/calmodulin-dependent protein kinase II (CaMKII); Intracerebral hemorrhage (ICH); Necroptosis; Receptor-interacting protein 3 (RIP3).

Products

Cat. No.

Product Name

Description

Target

Research Area
HY-15465

KN-93

99.42%, CaMK II Inhibitor‎

CaMK; Autophagy

Cancer

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