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环王巴明加剧大鼠脑缺血再灌注损伤
王莉,余萍萍,唐凡人,周露玲,曾立,宋晓松,陈吉祥,杨琴
0
(重庆医科大学附属第一医院神经内科, 重庆 400016
*通信作者)
摘要:
目的 探讨环王巴明(Cyc)对大鼠脑缺血再灌注损伤的影响。方法 60只SD雄性大鼠随机分成假手术组(Sham组)、脑缺血再灌注对照组(Con组)和脑缺血再灌注Cyc干预组(Cyc组),每组20只。于脑缺血再灌注后3 h腹腔注射Cyc(10 mg/kg)或无水乙醇,连用7 d。采用Longa评分法行脑缺血后1 d和14 d神经功能缺损评分。脑缺血24 h时,干湿质量法检测脑含水量,氯化三苯基四氮唑(TTC)染色测脑梗死体积,H-E染色观察病理改变,原位末端标记(TUNEL)法检测细胞凋亡。免疫化学法检测缺血后24 h NeuN、caspase-3蛋白及14 d时胶质纤维酸性蛋白(GFAP)表达。结果 Cyc和Con组大鼠神经功能缺损评分、脑含水量、脑梗死体积、TUNEL阳性细胞计数、caspase-3和GFAP蛋白表达均高于假手术组(P<0.05),且Cyc组较Con组更高(P<0.05)。Cyc和Con组NeuN蛋白表达低于假手术组(P<0.05),且Cyc组较Con组更低(P<0.05)。组织病理见Cyc组细胞和间质水肿、神经细胞变形、核固缩、细胞坏死等重于Con组。结论 Cyc可加剧大鼠脑缺血再灌注损伤。
关键词:  环王巴明  脑缺血  再灌注损伤  caspase-3  胶质纤维酸性蛋白
DOI:10.16781/j.0258-879x.2016.06.0677
投稿时间:2015-09-09修订日期:2016-03-01
基金项目:国家自然科学基金面上项目(81071119),卫生和计划生育委员会国家临床重点专科建设项目[卫办医政函 (2012) 649号].
Cyclopamine aggravates rat cerebral ischemia-reperfusion injury
WANG Li,YU Ping-ping,TANG Fan-ren,ZHOU Lu-ling,ZENG Li,SONG Xiao-song,CHEN Ji-xiang,YANG Qin
(Department of Neurology, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
*Corresponding author)
Abstract:
Objective To explore the effect of cyclopamine (Cyc) on cerebral ischemia-reperfusion injury in rats. Methods Sixty male Sprague-Dawley (SD) rats were randomly divided into 3 groups (n=20): sham operation group, cerebral ischemia-reperfusion group (Con group) and cerebral ischemia-reperfusion+Cyc group (Cyc group). Cyc (10 mg/kg) or absolute ethyl alcohol was intraperitoneally injected in animals 3 h after cerebral ischemia for 7 d. Neurological deficit was assessed by Longa scale on day 1 and 14 after cerebral ischemia. At 24 h after cerebral ischemia, the cerebral water content, cerebral infaction area, pathological changes and cell apoptosis were evaluated by dry-wet method, 2,3,5-triphenyltetrazolium (TTC) staining, H-E staining and TUNEL method, respectively. Immunochemical method was used to examine the protein expression of NeuN and caspase-3 at 24 h after ischemia and glial fibrillary acidic protein (GFAP) expression on day 14 after ischemia. Results Neurological deficit score, cerebral water content, cerebral infaction area, TUNEL positive cell counting, protein levels of caspase-3 and GFAP in Cyc and Con groups were significantly higher than those in sham operation group (P<0.05), and the above parameters in Cyc group were also significantly higher than those in Con group (P<0.05). NeuN protein expressions in Cyc and Con groups were significantly lower than those in sham operation group (P<0.05), and NeuN protein in Cyc group was also significantly lower than that in Con group (P<0.05). Cellular and interstitial edema, neurocyte deformation, pyknosis and necrocytosis were more severe in Cyc group than in Con group. Conclusion Cyclopamine can aggravate rat cerebral ischemia-reperfusion injury.
Key words:  cyclopamine  cerebral ischemia  reperfusion injuries  caspase-3  glial fibrillary acidic protein