Please wait a minute...
浙江大学学报（医学版）
浙江大学学报（医学版）  2018, Vol. 47 Issue (1): 19-26    DOI: 10.3785/j.issn.1008-9292.2018.02.03
专题报道     
半胱氨酰白三烯受体拮抗剂对全脑缺血再灌注慢性损伤的作用
王浩1(),郭红刚2,楼琦2,石巧娟2,*()
1. 浙江省立同德医院神经内科, 浙江 杭州 310012
2. 浙江省医学科学院实验动物中心, 浙江 杭州 310013
Effects of cysteinyl leukotrienes receptor antagonists on chronic brain injury after global cerebral ischemia/reperfusion
WANG Hao1(),GUO Honggang2,LOU Qi2,SHI Qiaojuan2,*()
1. Department of Neurology, Tongde Hospital of Zhejiang Province, Hangzhou 310012, China
2. Laboratory Animal Center, Zhejiang Academy of Medical Sciences, Hangzhou 310013, China
 全文: PDF(1117 KB)   HTML( 11 )
摘要：

目的: 研究半胱氨酰白三烯受体（CysLTR）拮抗剂普鲁司特和HAMI 3379对全脑缺血再灌注慢性损伤的保护作用及相关作用机制。方法: 40只体质量为45~65 g的清洁级健康雄性长爪沙鼠分为手术对照组、模型对照组、普鲁司特组和HAMI 3379组，每组10只。采用结扎双侧颈总动脉10 min再灌注法制作全脑缺血再灌注损伤模型。普鲁司特组和HAMI 3379组于术前30 min和术后30 min、4 h、12 h分别腹腔注射普鲁司特和HAMI 3379，第二天起每天分别给药一次，连续给药5 d。于全脑缺血再灌注24 h和14 d时对各组的神经症状和功能进行评分；尼氏染色法观察再灌注14 d时各组大脑皮层神经元的形态和数量；免疫组织化学染色检测再灌注14 d时各组大脑皮层小胶质细胞和星形胶质细胞激活情况。结果: 30只长爪沙鼠中，21只造模成功，其中模型对照组7只，普鲁司特组6只，HAMI 3378组8只。与模型对照组比较，普鲁司特组和HAMI 3379组术后24 h神经症状评分降低（均P < 0.01），术后14 d普鲁司特组和HAMI 3379组的神经症状评分较模型对照组亦有改善的趋势，但差异均无统计学意义（均P > 0.05）；普鲁司特组和HAMI 3379组在这两个时间点的神经功能评分均高于模型对照组（P < 0.05或P < 0.01）。普鲁司特组和HAMI 3379组大脑皮层神经元损伤较模型对照组减轻，存活神经元密度与模型对照组差异有统计学意义（均P < 0.01）。普鲁司特组和HAMI 3379组大脑皮层小胶质细胞和星形胶质细胞增生情况较模型对照组改善（均P < 0.01）。结论: 普鲁司特和HAMI 3379对长爪沙鼠全脑缺血慢性损伤模型具有较持久的神经保护作用。
关键词： 白三烯拮抗剂/药理学;  半胱氨酸;  沙丁胺醇/治疗应用;  脑缺血/病理生理学;  再灌注损伤/药物作用;  神经元/药物作用;  疾病模型, 动物;  沙鼠亚科    
Abstract:

Objective: To investigate the effects of cysteinyl leukotrienes receptor (CysLTR) antagonists on global cerebral ischemia/reperfusion (CI/R) injury in gerbils, and to explore its mechanism. Methods: Totally 40 gerbils weighting 45-65 g were randomized into sham, saline, Pranlukast and HAMI 3379 groups with 10 animals in each. The CI/R model was established in gerbils by bilateral common carotid occlusion for 10 min followed by reperfusion. After ischemia, the CysLTR antagonists Pranlukast (0.1 mg/kg) and HAMI 3379 (0.1 mg/kg) were injected intraperitoneally for 5 consecutive days in the last two groups, while the former two groups were injected with saline only (10 mL/kg). After 24 h or 14 d reperfusion, neurological deficit score was evaluated and the behavioral dysfunction was assessed, respectively. And 14 d after reperfusion, the neuron morphology of cerebral cortex was observed in brain sections stained with Cresyl violet. In addition, the Iba-1 (microgila) and GFAP (astrocyte) positive cells in cerebral cortex were observed by using immunohistochemitry method. Results: CI/R models were successfully established in 21 out of 30 gerbils with 7 in saline group, 6 in Pranlukast group, and 8 in HAMI 3379 group. Compared with saline group, Pranlukast and HAMI 3379 significantly attenuated neurological deficits, improved the behavioral function 24 h after reperfusion(all P < 0.01); Pranlukast and HAMI 3379 also significantly improved the behavioral function 14 days after reperfusion(P < 0.05 or P < 0.01). Compared with saline group, the neurological symptom scores in Pranlukast and HAMI 3379 groups presented a trend of amelioration 14 d after reperfusion, but it was not significant(P > 0.05). In addition, Pranlukast and HAMI 3379 also inhibited the neuron loss and injury, suppressed microgila and astrocyte activation 14 d after reperfusion(all P < 0.01). Conclusion: CysLTR antagonists Pranlukast and HAMI 3379 have long-term neuroprotective effect on chronic brain injury induced by global cerebral ischemia/reperfusion in gerbils.
Key words: Leukotriene antagonists/pharmacology    Cysteine    Albuterol/therapeutic use    Brain ischemia/physiopathology    Reperfusion injury/drug effects    Neurons/drug effects    Disease models, animal    Gerbillinae
收稿日期: 2017-07-07 出版日期: 2018-06-12
CLC:  R743  
基金资助: 国家自然科学基金(31301933);浙江省自然科学基金(LQ15H090005);浙江省科技计划项目(2014C37011);浙江省医药卫生科技计划(2015117570, 201477310)
通讯作者: 石巧娟     E-mail: wangh8234@163.com;shiqiaojuan@163.com
作者简介: 王浩(1982-), 男, 硕士, 副主任医师, 主要从事脑血管病发病机制研究; E-mail:wangh8234@163.com; https://orcid.org/0000-0002-0802-2226
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　
王浩
郭红刚
楼琦
石巧娟

引用本文:

王浩,郭红刚,楼琦,石巧娟. 半胱氨酰白三烯受体拮抗剂对全脑缺血再灌注慢性损伤的作用[J]. 浙江大学学报（医学版）, 2018, 47(1): 19-26.

WANG Hao,GUO Honggang,LOU Qi,SHI Qiaojuan. Effects of cysteinyl leukotrienes receptor antagonists on chronic brain injury after global cerebral ischemia/reperfusion. J Zhejiang Univ (Med Sci), 2018, 47(1): 19-26.

链接本文:

http://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2018.02.03    或    http://www.zjujournals.com/med/CN/Y2018/V47/I1/19

图 1  长爪沙鼠全脑缺血过程中脑血流监测结果
图 2  普鲁司特和HAMI 3379对全脑缺血再灌注模型神经症状和神经功能的影响
图 3  普鲁司特和HAMI 3379可改善全脑缺血再灌注模型大脑皮层神经元损伤(尼氏染色)
图 4  普鲁司特和HAMI 3379可抑制全脑缺血再灌注模型大脑皮层小胶质细胞激活(免疫组织化学染色)
图 5  普鲁司特和HAMI 3379可抑制全脑缺血再灌注模型大脑皮层星形胶质细胞激活(免疫组织化学染色)
1 IADECOLA C , ALEXANDER M . Cerebral ischemia and inflammation[J]. Curr Opin Neurol, 2001, 14 (1): 89- 94
doi: 10.1097/00019052-200102000-00014
2 SILVER J , MILLER J H . Regeneration beyond the glial scar[J]. Nat Rev Neurosci, 2004, 5 (2): 146- 156
doi: 10.1038/nrn1326
3 FAWCETT J W , ASHER R A . The glial scar and central nervous system repair[J]. Brain Res Bull, 1999, 49 (6): 377- 391
doi: 10.1016/S0361-9230(99)00072-6
4 魏尔清 . 半胱氨酰白三烯受体调节脑内炎症病变的研究展望[J]. 浙江大学学报(医学版), 2011, 40 (2): 117- 122
WEI Erqing . Prospects for research in regulation of brain inflammation by cysteinyl leukotriene receptors[J]. Journal of Zhejiang University(Medical Sciences), 2011, 40 (2): 117- 122
5 NI N C , YAN D , BALLANTYNE L L et al. A selective cysteinyl leukotriene receptor 2 antagonist blocks myocardial ischemia/reperfusion injury and vascular permeability in mice[J]. J Pharmacol Exp Ther, 2011, 339 (3): 768- 778
doi: 10.1124/jpet.111.186031
6 WUNDER F , TINEL H , KAST R et al. Pharmacological characterization of the first potent and selective antagonist at the cysteinyl leukotriene 2(CysLT(2)) receptor[J]. Br J Pharmacol, 2010, 160 (2): 399- 409
doi: 10.1111/j.1476-5381.2010.00730.x
7 张晶, 郭继东, 邢淑华 et al. 褪黑素对沙土鼠脑缺血再灌注损伤的保护作用[J]. 药学学报, 2002, 37 (5): 329- 333
ZHANG Jing , GUO Jidong , XING Shuhua et al. The protective effects of melatonin on global cerebral ischemia-reperfusion injury in gerbils[J]. Acta Pharmaceutica Sinica, 2002, 37 (5): 329- 333
8 ZHANG L H , WEI E Q . Neuroprotective effect of ONO-1078, a leukotriene receptor antagonist, on transient global cerebral ischemia in rats[J]. Acta Pharmacol Sin, 2003, 24 (12): 1241- 1247
9 ZHANG W P , WEI E Q , MEI R H et al. Neuroprotective effect of ONO-1078, a leukotriene receptor antagonist, on focal cerebral ischemia in rats[J]. Acta Pharmacol Sin, 2002, 23 (10): 871- 877
10 CHU L S , WEI E Q , YU G L et al. Pranlukast reduces neutrophil but not macrophage/microglial accumulation in brain after focal cerebral ischemia in mice[J]. Acta Pharmacol Sin, 2006, 27 (3): 282- 288
doi: 10.1111/aphs.2006.27.issue-3
11 QIAN X D , WEI E Q , ZHANG L et al. Pranlukast, a cysteinyl leukotriene receptor 1 antagonist, protects mice against brain cold injury[J]. Eur J Pharmacol, 2006, 549 (1-3): 35- 40
doi: 10.1016/j.ejphar.2006.07.056
12 SHI W Z , QI L L , FANG S H et al. Aggravated chronic brain injury after focal cerebral ischemia in aquaporin-4-deficient mice[J]. Neurosci Lett, 2012, 520 (1): 121- 125
doi: 10.1016/j.neulet.2012.05.052
13 YUZAWA I , YAMADA M , FUJⅡ K . An oral administration of cilostazol before focal ischemia reduces the infarct volume with delayed cerebral blood flow increase in rats[J]. J Stroke Cerebrovasc Dis, 2008, 17 (5): 281- 286
doi: 10.1016/j.jstrokecerebrovasdis.2008.03.007
14 CHOI J H , YOO K Y , LEE C H et al. Comparison of neurogenesis in the dentate gyrus between the adult and aged gerbil following transient global cerebral ischemia[J]. Neurochem Res, 2012, 37 (4): 802- 810
doi: 10.1007/s11064-011-0675-z
15 KIM J M , KIM S , KIM D H et al. Neuroprotective effect of forsythiaside against transient cerebral global ischemia in gerbil[J]. Eur J Pharmacol, 2011, 660 (2-3): 326- 333
doi: 10.1016/j.ejphar.2011.03.051
16 ICHIKI H , KUROIWA T , TANIGUCHI I et al. Delayed recovery of cortical auditory evoked potentials and blood flow precede cortical neuronal death after transient cerebral ischemia in gerbils[J]. J Med Dent Sci, 1998, 45 (4): 221- 232
17 HOSOMI N , TSUDA Y , ICHIHARA S I et al. Duration threshold of induced hypertension on cerebral blood flow, energy metabolism, and edema after transient forebrain ischemia in gerbils[J]. J Cereb Blood Flow Metab, 1996, 16 (6): 1224- 1229
doi: 10.1097/00004647-199611000-00017
18 PASCHEN W , MIES G , HOSSMANN K A . Threshold relationship between cerebral blood flow, glucose utilization, and energy metabolites during development of stroke in gerbils[J]. Exp Neurol, 1992, 117 (3): 325- 333
doi: 10.1016/0014-4886(92)90142-D
19 KAUSHIK A , JAYANT R D , NAIR M . Advancements in nano-enabled therapeutics for neuroHIV management[J]. Int J Nanomedicine, 2016, 11 4317- 4325
doi: 10.2147/IJN
20 R?HL C , LUCIUS R , SIEVERS J . The effect of activated microglia on astrogliosis parameters in astrocyte cultures[J]. Brain Res, 2007, 1129 (1): 43- 52
21 BARRON K D . The microglial cell. A historical review[J]. J Neurol Sci, 1995, 34 Suppl 57- 68
22 DIPROSPERO N A , MEINERS S , GELLER H M . Inflammatory cytokines interact to modulate extracellular matrix and astrocytic support of neurite outgrowth[J]. Exp Neurol, 1997, 148 (2): 628- 639
doi: 10.1006/exnr.1997.6700
23 HUANG X Q , ZHANG X Y , WANG X R et al. Transforming growth factor β1-induced astrocyte migration is mediated in part by activating 5-lipoxygenase and cysteinyl leukotriene receptor 1[J]. J Neuroinflammation, 2012, 9 145
24 ZHANG X Y , WANG X R , XU D M et al. HAMI 3379, a CysLT2 receptor antagonist, attenuates ischemia-like neuronal injury by inhibiting microglial activation[J]. J Pharmacol Exp Ther, 2013, 346 (2): 328- 341
doi: 10.1124/jpet.113.203604
25 Stroke Therapy Academic Industry Roundtable . Recommendations for standards regarding preclinical neuroprotective and restorative drug development[J]. Stroke, 1999, 30 (12): 2752- 2758
doi: 10.1161/01.STR.30.12.2752
26 RICHARD G A , ODERGREN T , ASHWOOD T . Animal models of stroke:do they have value for discovering neuroprotective agents?[J]. Trends Pharmacol Sci, 2003, 24 (8): 402- 408
doi: 10.1016/S0165-6147(03)00192-5
[1] 王晓蓉,卢韵碧,张纬萍,魏尔清,方三华. 半胱氨酰白三烯受体对小鼠小胶质细胞吞噬功能的调节作用[J]. 浙江大学学报（医学版）, 2018, 47(1): 10-18.
[2] 钟根龙 等. 血清叶酸水平与脑白质高信号严重程度及伴发脑微出血的相关性[J]. 浙江大学学报（医学版）, 2017, 46(4): 390-396.
[3] 朱惠惠, 赵西宝, 胡未伟, 陈玮琳. 泛素特异性蛋白酶在抗病毒感染免疫中的作用研究进展[J]. 浙江大学学报（医学版）, 2015, 44(5): 578-583.
[4] 缪成贵, 周国梁, 秦梅颂, 陈建中, 李成凤, 张兵. 禹州漏芦总黄酮对类风湿关节炎大鼠治疗机制研究[J]. 浙江大学学报（医学版）, 2015, 44(1): 43-48.
[5] 王浩, 耿赵铭, 胡智伟, 王舒燕, 赵冰. 丹皮酚抑制帕金森病模型细胞凋亡的作用[J]. 浙江大学学报（医学版）, 2015, 44(1): 30-36.
[6] 韩晓瑜，章璐，刘志娴，等. 白三烯D4在体外对A549肺泡上皮细胞增殖及迁移的影响[J]. 浙江大学学报（医学版）, 2014, 43(3): 287-292.
[7] 王晓蓉，张霞燕，徐栋敏，等. 组蛋白去乙酰基酶抑制剂NL101对大鼠神经元的作用[J]. 浙江大学学报（医学版）, 2014, 43(3): 265-272.
[8] 徐栋敏，张霞燕，王晓蓉，等. 孟鲁司特和HAMI 3379对体外大鼠皮层神经元[J]. 浙江大学学报（医学版）, 2014, 43(3): 257-264.
[9] 张壮, 骆江云, 黄静, 刘志娴, 方三华, 张纬萍, 魏尔清, 卢韵碧. 白三烯D4体外诱导小胶质细胞激活的作用[J]. 浙江大学学报（医学版）, 2013, 42(3): 253-260.
[10] 张壮, 魏尔清, 卢韵碧. GPR17在中枢神经系统损伤修复中的作用研究进展[J]. 浙江大学学报（医学版）, 2013, 42(3): 355-359.
[11] . 天花粉多糖诱导人乳腺癌MCF-7细胞凋亡及其Caspase-3和Caspase-8活化对凋亡的影响[J]. 浙江大学学报（医学版）, 2012, 41(5): 527-534.
[12] . 人参皂苷Rg1经线粒体通路抗Aβ25-35致原代大鼠皮层神经元凋亡[J]. 浙江大学学报（医学版）, 2012, 41(4): 393-401.
[13] . 半胱氨酰白三烯受体1拮抗剂孟鲁司特对缺血性损伤后神经元形态的作用[J]. 浙江大学学报（医学版）, 2012, 41(3): 259-266.
[14] .
半胱氨酰白三烯受体1参与鱼藤酮诱导PC12细胞损伤的调节[J]. 浙江大学学报（医学版）, 2012, 41(2): 139-145.
[15] . 幽门螺杆菌感染与脑梗死患者同型半胱氨酸的关系[J]. 浙江大学学报（医学版）, 2012, 41(1): 89-92.