Hydroxychloroquine reduces cerebral ischemia reperfusion
injury by promoting ERK1/2 phosphorylation
HU Xiu-lan1, SHAO Dong-hua2, MA Xiao-dong2, LUO Hong2, XIE Yun-bin2, XIA Yan2
(1. Department of Intensive Care Unit, 2. Department of Anesthesiology, Affiliated People′s Hospital of Jiangsu University, Zhenjiang Jiangsu 212002, China)
Abstract：Objective: To evaluate the attenuation of hydroxychloroquine(HCQ) for cerebral ischemia reperfusion(I/R) injury induced by transient middle cerebral artery occlusion(MCAO) in rats and potential mechanisms. Methods: A total of 72 healthy male SD rats were randomly divided into 6 groups (n=12):control group, ischemiareperfusion group (group I/R), low dose HCQ pretreatment group (group HCQ250), medium dose HCQ pretreatment group (group HCQ500),high dose HCQ pretreatment group (group HCQ1000) and extracellular regulated protein kinase 1/2(ERK1/2) inhibitor U0126 pretreatment group (group U0126). MCAO was used to establish rat ischemic stroke model. Control group was the same as group I/R, but not occluded the middle cerebral artery; group I/R was reperfused after MCAO for 2 h; group HCQ250, group HCQ500, group HCQ1000 and group U0126 were injected with 8 μL 250 μmol/L, 500 μmol/L, 1 000 μmol/L and 1 000 μmol/L HCQ by lateral ventricle (LV) at 24 h, 48 h and 72 h, respectively, before MCAO, and U0126 group was injected with 8 μL of 1 000 μmol/L U0126 after 12 h each injection of HCQ. Brain tissues were collected by decollation after reperfusion 6 h, the expression of ERK1/2, pERK1/2, antiapoptotic factor Bcl2,proapoptotic factor cleaved caspase3,pCREB and pAkt was detected by Western blotting in ischemic penumbra. Neurological deficit score was measured and followed infarct volume was detection by TTC staining 24 h after reperfusion. Results: Compared with control group, neurological deficit score was reduced, infarct volume was magnified, the expression of pERK1/2, cleaved caspase3, pCREB, pAkt was increased, Bcl2 expression was decreased in group I/R , group HCQ250, group HCQ500, group HCQ1000 and group U0126(P<0.05). Compared with group I/R, neurological deficit score was enlarged, infarct volume was reduced, the expression of pERK1/2, Bcl2, pCREB, pAkt was increased, the expression of cleaved caspase3 was decreased in group HCQ1000 (P<0.05). However, compared with group HCQ1000, neurological deficit score was reduced, infarct volume was magnified, the expression of pERK1/2, Bcl2, pCREB, pAkt was decreased, the expression of cleaved caspase3 was increased in group U0126(P<0.05). Conclusion: Hydroxychloroquine may promote the expression of pERK1/2 and protects agaist cerebral ischemia through reperfusion injury through CREB/Akt signaling way in rats.
［1］Writing Group Members, Mozaffarian D, Benjamin EJ,et al.Executive summary: heart disease and stroke statistics—2016 update:a report from the American Heart Association\[J\].Circulation,2016,133(4):447-454.［2］Li M,Feng B,Wang L,et al.Tollip is a critical mediator of cerebral ischaemiareperfusion injury\[J\]. J Pathol,2015,237(2):249-262.［3］MartinezLopez N,Athonvarangkul D,Mishall P,et al.Autophagy proteins regulate ERK phosphorylation\[J\]. Nat Commun,2013,4:2799.［4］Li L,Xiong Y,Qu Y,et al.The requirement of extracellular signalrelated protein kinase pathway in the activation of hypoxia inducible factor 1 α in the developing rat brain after hypoxiaischemia\[J\]. Acta Neuropathol,2008,115(3):297-303.［5］Liu L,Zhang R,Liu K,et al.Tissue kallikrein protects cortical neurons against in vitro ischemiaacidosis /reperfusioninduced injury through the ERK1/2 pathway\[J\].Exp Neurol,2009,219(2): 453-465.［6］Bourke L,Mccormick J,Taylor V,et al.Hydroxychloroquine protects against cardiac ischaemia/reperfusion injury in vivo via enhancement of ERK1/2 phosphorylation\[J\].PLoS One,2015,10(12):e143771.［7］Liang X, Hu Q, Li B, et al. Follistatinlike 1 attenuates apoptosis via discointeracting protein 2 homolog A/Akt pathway after middle cerebral artery occlusion in rats\[J\]. Stroke,2014,45(10):3048-3054.［8］Hommel JD, Sears RM, Georgescu D, et al. Local gene knockdown in the brain using viralmediated RNA interference\[J\]. Nat Med,2003,9(12):1539-1544.［9］Mebratu Y,Tesfaigzi Y.How ERK1/2 activation controls cell proliferation and cell death:Is subcellular localization the answer?\[J\].Cell Cycle,2009,8(8):1168-1175.［10］Wang Y,Han R,Zuo Z.Dexmedetomidine posttreatment induces neuroprotection via activation of extracellular signal regulated kinase in rats with subarachnoid haemorrhage\[J\].Br J Anaesth,2016,116 (3):384-392.［11］Lee SJ, Silverman E, Bargman JM , et al.The role of antimalarial agents in the treatment of SLE and lupus nephritis\[J\]. Nat Rev Nephrol,2011,7: 718-729.［12］Barth S, Glick D, Macleod KF,et al. Autophagy: assays and artifacts\[J\].J Pathol,2010, 221:117-124.［13］Johnson R, Charnley J. Hydroxychloroquine in prophylaxis of pulmonary embolism following hip arthroplasty\[J\].Clin Orthop Relat Res, 1979(144):174-177.［14］Dworkin S,Mantamadiotis T.Targeting CREB signaling in neurogenesis\[J\].Expert Opin Ther Targets,2010,14(8):869-879.
2019, Vol. 29 
