Rapamycin treatment starting at 24 h after cerebral ischemia/reperfusion exhibits protective effect on brain injury in rats

doi:10.3785/j.issn.1008-9292.2018.10.01

J Zhejiang Univ (Med Sci)

2018, Vol. 47

Issue (5): 443-449 DOI: 10.3785/j.issn.1008-9292.2018.10.01

Rapamycin treatment starting at 24 h after cerebral ischemia/reperfusion exhibits protective effect on brain injury in rats

LIANG Gang(

),NIU Yumiao,LI Yihan,Wei Anyi,DONG Jingyin*(

),ZENG Linghui*(

)

School of Medicine, Zhejiang University City College, Hangzhou 310015, China

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Abstract

Objective: To investigate whether rapamycin treatment starting at 24 h after cerebral ischemia/reperfusion(I/R) has protective effect on brain injury in rats. Methods: The rat I/R model was established by middle cerebral artery occlusion according to Longa's method. A total of 104 Sprague Dawley rats were randomly divided into sham group, model group, and rapamycin-treated groups (6 h or 24 h after modeling). Neurological function was assessed with neurological severity score (NSS). Triphenyl tetrazolium chloride (TTC) staining and Fluoro-Jade B (FJB) staining were used to examine the infarct volume and neuronal apoptosis, respectively. The expression of p-S6 protein in mTOR signaling pathway was detected by Western blot analysis. Results: Compared with sham group, NSS of the model group was significantly increased and TTC staining indicated obvious infarct area (all P < 0.01). Furthermore, significantly increased number of FJB-positive cells and p-S6 expression in the penumbra area were shown in the model group (all P < 0.01). Compared with the model group, both rapamycin-treated groups demonstrated decreased NSS, infarction volume and FJB positive cells as well as p-S6 expression in the penumbra area (P < 0.05 or P < 0.01). There was no significant difference between the groups of rapamycin administrated 6 h and 24 h after modeling (all P > 0.05). Conclusion: Rapamycin treatment starting at 24 h after I/R exhibits protective effect on brain injury in rats.

Key words： Brain ischemia/drug therapy Reperfusion injury/drug therapy Neurons Apoptosis Protein kinases/physiology Sirolimus/pharmacology Signal transduction

Received: 12 September 2018 Published: 23 January 2019

CLC:

R743

Corresponding Authors: DONG Jingyin,ZENG Linghui

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	LIANG Gang
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Cite this article:

LIANG Gang, NIU Yumiao, LI Yihan, Wei Anyi, DONG Jingyin, ZENG Linghui. Rapamycin treatment starting at 24 h after cerebral ischemia/reperfusion exhibits protective effect on brain injury in rats. J Zhejiang Univ (Med Sci), 2018, 47(5): 443-449.

URL:

http://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2018.10.01 OR http://www.zjujournals.com/med/Y2018/V47/I5/443

雷帕霉素在大鼠局灶性脑缺血再灌注后24 h给药对脑损伤的保护作用

目的: 探讨雷帕霉素在大鼠局灶性脑缺血再灌注后24 h给药对脑损伤的保护作用。方法: 104只Sprague Dawley大鼠随机分为手术对照组、模型对照组、建模后6 h雷帕霉素给药组（6 h给药组）和建模后24 h雷帕霉素给药组（24 h给药组）。采用Longa法建立大鼠大脑中动脉缺血再灌注模型。采用神经功能损伤程度评分（NSS）对大鼠进行神经功能评分；氯化三苯基四氮唑（TTC）染色法检测各组大鼠脑梗死体积；Fluoro-Jade B（FJB）染色法检测大鼠脑组织神经元凋亡；蛋白质印迹法检测各组大鼠mTOR信号通路磷酸化S6蛋白表达。结果: 与手术对照组比较，模型对照组大鼠NSS升高，脑梗死体积增加，FJB阳性细胞增多，半暗带磷酸化S6蛋白表达量增加（均P < 0.01）；与模型对照组比较，6 h和24 h给药组大鼠NSS均降低，脑梗死体积缩小，FJB阳性细胞减少，半暗带磷酸化S6蛋白表达量减少（均P < 0.05或P < 0.01），且24 h给药组各项指标变化与6 h给药组差异均无统计学意义（均P > 0.05）。结论: 缺血再灌注后24 h给予雷帕霉素对大鼠局灶性脑缺血再灌注所致脑损伤仍有保护作用。

关键词： 脑缺血/药物疗法, 再灌注损伤/药物疗法, 神经元, 细胞凋亡, 蛋白激酶类/生理学, 西罗莫司/药理学, 信号传导

Tab 1 Neurological severity score^[15]

Fig 1 Rapamycin administered at different stages inhibited neurological injury in I/R rats (n=10)

Fig 2 Rapamycin administered at different stages reduced cerebral infarction volume in I/R rats

Fig 3 Rapamycin administered at different stages reduced neuronal apoptosis in I/R rats

Fig 4 Phosphorylation of S6 in I/R rats (n=6)


[1]	GAO H J , LIU P F , LI P W et al. Ligustrazine monomer against cerebral ischemia/reperfusion injury[J]. Neural Regen Res, 2015, 10 (5): 832- 840 doi: 10.4103/1673-5374.156991

[2]	PAN Y, CAI X, HUO X, et al. Cost-effectiveness of mechanical thrombectomy within 6 hours of acute ischaemic stroke in China[J/OL]. BMJ Open, 2018, 8(2): e018951.

[3]	CHONG Z Z , YAO Q Q , LI H H . The rationale of targeting mammalian target of rapamycin for ischemic stroke[J]. Cell Signal, 2013, 25 (7): 1598- 1607 doi: 10.1016/j.cellsig.2013.03.017

[4]	HACKE W , DONNAN G , FIESCHI C et al. Association of outcome with early stroke treatment:pooled analysis of ATLANTIS, ECASS, and NINDS rt-PA stroke trials[J]. Lancet, 2004, 363 (9411): 768- 774 doi: 10.1016/S0140-6736(04)15692-4

[5]	CHOI J H , PILE-SPELLMAN J . Reperfusion changes after stroke and practical approaches for neuroprotection[J]. Neuroimaging Clin N Am, 2018, 28 (4): 663- 682 doi: 10.1016/j.nic.2018.06.008

[6]	NARNE P , PANDEY V , PHANITHI P B . Role of nitric oxide and hydrogen sulfide in ischemic stroke and the emergent epigenetic underpinnings[J]. Mol Neurobiol, 2018,

[7]	HOU Y , WANG K , WAN W et al. Resveratrol provides neuroprotection by regulating the JAK2/STAT3/PI3K/AKT/mTOR pathway after stroke in rats[J]. Genes Dis, 2018, 5 (3): 245- 255 doi: 10.1016/j.gendis.2018.06.001

[8]	SHI G D , OUYANG Y P , SHI J G et al. PTEN deletion prevents ischemic brain injury by activating the mTOR signaling pathway[J]. Biochem Biophys Res Commun, 2011, 404 (4): 941- 945 doi: 10.1016/j.bbrc.2010.12.085

[9]	MAIESE K . Cutting through the complexities of mTOR for the treatment of stroke[J]. Curr Neurovasc Res, 2014, 11 (2): 177- 186 doi: 10.2174/1567202611666140408104831

[10]	FLETCHER L, EVANS T M, WATTS L T, et al. Rapamycin treatment improves neuron viability in an in vitro model of stroke[J/OL]. PLoS One, 2013, 8(7): e68281.

[11]	URBANEK T , KUCZMIK W , BASTA-KAIM A et al. Rapamycin induces of protective autophagy in vascular endothelial cells exposed to oxygen-glucose deprivation[J]. Brain Res, 2014, 1553:1- 11 doi: 10.1016/j.brainres.2014.01.017

[12]	张斌斌, 吴美玲, 刘露娜 et al. 抑制哺乳动物雷帕霉素靶蛋白信号通路对慢性脑缺血小鼠认知功能的改善和机制[J]. 浙江大学学报(医学版), 2017, 46 (4): 405- 411 ZHANG Binbin , WU Meiling , LIU Luna et al. Inhibiting mammalian target of rapamycin signaling pathway improves cognitive function in mice with chronic cerebral ischemia[J]. Journal of Zhejiang University(Medical Sciences), 2017, 46 (4): 405- 411

[13]	WU M , ZHANG H , KAI J et al. Rapamycin prevents cerebral stroke by modulating apoptosis and autophagy in penumbra in rats[J]. Ann Clin Transl Neurol, 2018, 5 (2): 138- 146

[14]	LONGA E Z , WEINSTEIN P R , CARLSON S et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke, 1989, 20 (1): 84- 91 doi: 10.1161/01.STR.20.1.84

[15]	CHEN J , SANBERG P R , LI Y et al. Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats[J]. Stroke, 2001, 32 (11): 2682- 2688 doi: 10.1161/hs1101.098367

[16]	WANG T, LI Y, WANG Y, et al. Lycium barbarum polysaccharide prevents focal cerebral ischemic injury by inhibiting neuronal apoptosis in mice[J/OL]. PLoS One, 2014, 9(3): e90780.

[17]	冯春燕, 樊小农, 张春红 et al. 氯化三苯四氮唑染色后梗死灶定量分析方法[J]. 生物工程学杂志, 2009, 26 (6): 1363- 1366 FENG Chunyan , FAN Xiaonong , ZHANG Chunhong et al. On the quantitative analysis of focal ischemic cerebral infarction by TTC staining[J]. Journal of Biomedical Engineering, 2009, 26 (6): 1363- 1366

[18]	ASHWAL S , TONE B , TIAN H R et al. Core and penumbral nitric oxide synthase activity during cerebral ischemia and reperfusion[J]. Stroke, 1998, 29 (5): 1037- 1046 doi: 10.1161/01.STR.29.5.1037

[19]	孙明, 赵育梅, 徐超 . 局灶性脑缺血再灌注损伤核心区及半暗带的组织学定位[J]. 中华神经外科杂志, 2003, 19 (4): 259- 262 SUN Ming , ZHAO Yumei , XU Chao . Histological location of core and penumbra of cortex and striatum underwent focal cerebral ischemia and reperfusion[J]. Chinese Journal of Neurosurgery, 2003, 19 (4): 259- 262 doi: 10.3760/j.issn:1001-2346.2003.04.005

[20]	刘震乾, 张清秀, 魏秀娥 et al. 小鼠脑缺血再灌注模型的优化及其效果评价[J]. 东南大学学报(医学版), 2017, 36 (3): 328- 332 LIU Zhenqian , ZHANG Qingxiu , WEI Xiu'er et al. Optimization and evaluation of cerebral ischemia reperfusion model in mice[J]. Journal of Southeast University(Medical Science Edition), 2017, 36 (3): 328- 332 doi: 10.3969/j.issn.1671-6264.2017.03.004

[21]	WALKER C L, WALKER M J, LIU N K, et al. Systemic bisperoxovanadium activates Akt/mTOR, reduces autophagy, and enhances recovery following cervical spinal cord injury[J/OL]. PLoS One, 2012, 7(1): e30012.

[22]	KOH S H , LO E H . The role of the PI3K pathway in the regeneration of the damaged brain by neural stem cells after cerebral infarction[J]. J Clin Neurol, 2015, 11 (4): 297- 304 doi: 10.3988/jcn.2015.11.4.297

[23]	KOROTCHKINA L G , LEONTIEVA O V , BUKREEVA E I et al. The choice between p53-induced senescence and quiescence is determined in part by the mTOR pathway[J]. Aging(Albany NY), 2010, 2 (6): 344- 352

[24]	KOH P O , CHO J H , WON C K et al. Estradiol attenuates the focal cerebral ischemic injury through mTOR/p70S6 kinase signaling pathway[J]. Neurosci Lett, 2008, 436 (1): 62- 66 doi: 10.1016/j.neulet.2008.02.061

[25]	HARA A , NIWA M , YOSHIMI N et al. Apoptotic cell death in vulnerable subpopulation of cerebellar granule cells[J]. Acta Neuropathol, 1997, 94 (5): 517- 518 doi: 10.1007/s004010050744

[26]	YIN L , YE S , CHEN Z et al. Rapamycin preconditioning attenuates transient focal cerebral ischemia/reperfusion injury in mice[J]. Int J Neurosci, 2012, 122 (12): 748- 756 doi: 10.3109/00207454.2012.721827

[27]	CHAUHAN A , SHARMA U , JAGANNATHAN N R et al. Rapamycin ameliorates brain metabolites alterations after transient focal ischemia in rats[J]. Eur J Pharmacol, 2015, 757:28- 33 doi: 10.1016/j.ejphar.2015.03.006

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