Ligustroflavone reduces necroptosis in rat brain after ischemic stroke through targeting RIPK1/RIPK3/MLKL

被引：0

作者：

ZHANG Yiyue ^{[1
]}

LIU Weining ^{[1
]}

LI Yueqi ^{[2
]}

LUO Xiuju ^{[2
]}

PENG Jun ^{[1
,3
]}

机构：

[1] Department of Pharmacology,Xiangya School of Pharmaceutical Sciences,Central South University

[2] Department of Laboratory Medicine,The Third Xiangya Hospital of Central South University

[3] Hunan Provincial Key Laboratory of Cardiovascular Research,Xiangya School of Pharmaceutical Sciences,Central South University

来源：

中国药理学与毒理学杂志 | 2019年 / 33卷 / 09期

关键词：

ligustroflavone; necroptosis; receptor-interacting protein kinase 1; receptor-interacting protein kinase 3; mixed lineage kinase domain-like;

D O I：

暂无

中图分类号：

R285.5 [中药实验药理];

学科分类号：

1008 ;

摘要：

OBJECTIVE To explore the effect of ligustroflavone on ischemic brain injury in stroke rat and the underlying mechanisms. METHODS A rat model of ischemic stroke was established by middle cerebral artery occlusion（MCAO）. Administration of ligustroflavone(10, 30, 60 mg·kg-1, ig) 15 min before ischemia, after which neurological deficit score and infarct volume were detected by longa score and TTC stain. The cell viability and necrosis rate of hypoxia-cultured PC12 cells（O2/N2/CO2, 1:94:5, 8 h） were evaluated by MTS and LDH release rate. Flow cytometry further verified the mortality rate of PC12 cells. Necroptosis-associated proteins（RIPK1, RIPK3 and MLKL/p-MLKL） were detected by Western blotting. The interaction between RIPK3 and RIPK1 or MLKL were confirmed by immunoprecipitation. Operating Environment（MOE） program demonstrated the possible combination of ligustroflavone with RIPK1, RIPK3 and MLKL. RESULTS Ischemic injury（increase in neurological deficit score and infarct volume） and upregulation of necroptosis-associated proteins were showed in rat MCAO model. Administration of ligustroflavone(30 mg·kg-1, ig) evidently improved neurological function, reduced infarct volume, and decreased the levels of necroptosis-associated proteins except the RIPK1. Consistently,hypoxia-cultured PC12 cells caused cellular injury（LDH release and necroposis） concomitant with up-regulation of necroptosis-associated proteins, and these phenomena were blocked in the presence of ligustroflavone(25 μmol·L-1)except the elevated RIPK1 levels. Using the Molecular Operating Environment（MOE） program, we identified RIPK1,RIPK3, and MLKL as potential targets of ligustroflavone. Further studies showed that the interaction between RIPK3 and RIPK1 or MLKL was significantly enhanced, which was blocked in the presence of ligustroflavone. CONCLUSION Ligustroflavone protects rat brain from ischemic injury, and its beneficial effect is related to the prevention of necroptosis through a mechanism involving targeting RIPK1, RIPK3, and/or MLKL.

引用

页码：703 / 703

页数：1