藏药诃子提取物对脑缺血再灌注损伤模型大鼠的保护作用及其机制研究
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篇名: 藏药诃子提取物对脑缺血再灌注损伤模型大鼠的保护作用及其机制研究
TITLE:
摘要: 目的:探讨藏药诃子提取物对脑缺血再灌注损伤模型大鼠的保护作用及其机制。方法:将40只雄性SD大鼠随机分为假手术组、模型组和诃子提取物低、中、高剂量组(20、50、100 mg/kg),每组8只。除假手术组外其余各组大鼠均采用改良Longa线栓法复制大脑中动脉阻塞再灌注模型。造模成功2 h后,各给药组大鼠均灌胃相应药物,假手术组和模型组大鼠均灌胃等体积生理盐水,每天1次,连续7 d。采用流式细胞仪测定各组大鼠外周血内皮祖细胞(EPCs)水平;采用改良Garcia法进行神经功能学评分;采用2,3,5-氯化三苯基四氮唑(TTC)染色法测定各组大鼠脑梗死体积百分比;采用苏木精-伊红染色法观察大鼠脑组织病理学变化;采用Weidner法测定大鼠脑组织梗死区域微血管密度(MVD);采用酶联免疫吸附法测定大鼠脑组织梗死区域血管内皮细胞生长因子(VEGF)、血管内皮细胞生长因子受体2(VEGFR2)、一氧化氮(NO)水平。结果:与假手术组比较,模型组大鼠外周血EPCs水平和神经功能学评分均显著下降(P<0.05或P<0.01);脑组织中可见明显的梗死区域,细胞排列紊乱,部分细胞核固缩、溶解,胞体缩小;脑梗死体积百分比显著提高(P<0.01);脑组织梗死区域MVD显著下降,VEGF、VEGFR2、NO水平均显著升高(P<0.05或P<0.01)。与模型组比较,诃子提取物中、高剂量组大鼠外周血EPCs水平和神经功能学评分均显著升高(P<0.05或P<0.01);脑组织梗死区域明显缩小,坏死细胞明显减少;脑梗死体积百分比显著下降(P<0.05);脑组织梗死区域MVD、VEGF、VEGFR2、NO水平均显著升高(P<0.05)。结论:诃子提取物对模型大鼠脑缺血再灌注损伤具有一定的保护作用,可促进其神经功能恢复和脑微血管形成,并可缩小其脑梗死体积;其机制可能与上调VEGF、VEGFR2、NO的表达有关。
ABSTRACT: OBJECTIVE: To investigate the protective effects and its mechanism of the extract from Tibetan medicine Terminalia chebula against cerebral ischemia-reperfusion injury model rats. METHODS: Totally 40 male SD rats were randomly divided into sham operation group, model group, T. chebula extract low-dose, medium-dose and high-dose groups (20, 50, 100 mg/kg), with 8 rats in each group. Except for sham operation group, middle cerebral artery occlusion reperfusion model was induced by modified Longa suture-occluded method in other groups. Two hours after modeling, administration groups were given relevant medicine intragastrically; sham operation group and model group were given constant volume of normal saline intragastrically, once a day, for consecutive 7 d. The levels of EPCs in peripheral blood of rats were detected by flow cytometry. The modified Garcia method was used for neurological function score. TTC staining was used to detect the volume percentage of cerebral infarction in rats. HE staining was used to observe the histopathological changes of brain tissue. The MVD of cerebral infarction area was detected by Weidner assay. The levels of VEGF, VEGFR2 and NO in cerebral infraction area were determined by ELISA. RESULTS: Compared with sham operation group, EPCs level in peripheral blood and neurological function score were decreased significantly in model group (P<0.05 or P<0.01). There were obvious infarction area, disordered arrangement of cells, pyknosis and lysis of some nucleus, cell body shrinkage, and significant increase in the volume percentage of cerebral infarction (P<0.01); MVD  decreased significantly in cerebral infarction area, while the levels of VEGF, VEGFR2 and NO increased significantly (P<0.05 or P<0.01). Compared with model group, EPCs levels in peripheral blood and neurological function scores were increased significantly in T. chebula extract medium-dose and high-dose groups (P<0.05 or P<0.01); infarction area of cerebral tissue was significantly reduced, necrotic cells were significantly reduced, and the volume percentage of cerebral infarction was significantly decreased (P<0.05); the levels of MVD, VEGF, VEGFR2 and NO in cerebral infarction area were significantly increased (P<0.05). CONCLUSIONS: T. chebula extract shows certain protective effect on cerebral ischemia-reperfusion injury, and can promote the recovery of neurological function and the formation of cerebral microvessels and reduce the volume of cerebral infarction. The mechanism of it may be associated with the expression up-regulation of VEGF, VEGFR2 and NO.
期刊: 2018年第29卷第24期
作者: 任丹君,丁一,丁笑刚,翟文杰,刘文星,刘天龙,李建光
AUTHORS: REN Danjun,DING Yi,DING Xiaogang,ZHAI Wenjie,LIU Wenxing,LIU Tianlong,LI Jianguang
关键字: 藏药;诃子;提取物;脑缺血再灌注损伤;大脑中动脉阻塞再灌注模型;大鼠;血管新生;保护作用;机制
KEYWORDS: Tibetan medicine; Terminalia chebula; Extract; Cerebral ischemia-reperfusion injury; Middle cerebral artery occlusion reperfusion model; Rats; Angiogenesis; Protective effect; Mechanism
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