【摘要】 目的探讨川蛭通络胶囊对小鼠局灶性脑缺血再灌注损伤的保护作用。方法采用ICR小鼠中动脉阻断(MCAO)制备小鼠局灶性脑缺血再灌注模型,将ICR小鼠随机分为假手术组(sham)、MCAO/R组、川蛭通络胶囊治疗组(CZTLC),每组15只。其中CZTLC组于手术前5 d,口服灌胃给药2次/d。缺血60 min、再灌注24 h对小鼠进行神经行为学评分,深度麻醉取小鼠脑组织进行病理学TTC、HE及甲苯胺蓝染色;超氧化物歧化酶(SOD)试剂盒、丙二醛(MDA)测试盒分别测定脑组织SOD及MDA含量;ELISA检测肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、IL-4及IL-10的表达变化;RT-PCR、Western Blot检测Bax、Bcl-2、Caspase-3基因及蛋白水平表达。结果 HE染色、尼氏染色显示,与假手术组相比,MCAO/R可明显诱导左侧皮层和海马CA1区神经元损伤(P>0.05)。模型组大鼠脑组织损伤侧主要表现神经元细胞体积缩小,胞浆空泡变性,细胞间排列疏松,海马、皮质的神经元大部分萎缩;细胞内尼氏小体数量减少,染色变浅。与模型组比较,观察到川蛭通络胶囊治疗组小鼠脑组织区域神经元细胞损伤较轻微,表现为神经细胞的紧密排列和细胞内尼氏小体数量增加。同时,与假手术组相比,药物处理组海马CA1和皮层的正常神经元数量显著减少。假手术组小鼠脑组织SOD活性明显高于模型组,MDA水平明显低于模型组(P>0.05);与模型组相比,川蛭通络胶囊组小鼠脑组织中SOD活性显著升高,MDA水平显著降低(P>0.05)。模型组小鼠脑匀浆TNF-α、IL-1β及IL-6含量较假手术组明显升高,而IL-10含量明显降低(P<0.05)。CZTLC干预组小鼠脑匀浆TNF-α、IL-1β及IL-6含量较模型组明显降低,而IL-10含量明显升高(P<0.05)。CZTLC可显著下调凋亡蛋白caspase-3及Bax在基因水平及蛋白水平的表达,同时其亦明显上调Bcl-2的表达,与CAMO/R组进行对比差异有统计学意义(P<0.05)。结论 CZTLC在小鼠脑缺血再灌注损伤中具有明确的保护作用,其机制与改善炎性反应及抑制细胞凋亡有关。更多还原

【Abstract】 Objective To investigate the protective effects Chuanzhi Tongluo capsule(CZTLC) on ischemia-reperfusion induced brain injury in mice.Methods The ischemia-reperfusion injury(ICR) mice models were established by middle-cerebral-artery-occlusion(MCAO) surgery.The mice with ICR were randomly divided into three groups: sham operation group, MCAO/R group and CZTLC treatment group, with 15 mice in each group.The mice in CZTLC group were orally administrated twice a day for 5 days before MCAO.The neurological function scores were measured at 1 h after ischemia and 24 h after reperfusion.And the brain tissues of mice were taken out for pathological TTC,HE and toluidine blue staining.The levels of SOD,MDA, IL-1β,TNF-α,IL-6,IL-10 in cerebral tissues of mice were detected by ELISA.Western blot and quantitative reverse transcription polymerase chain reaction(qRT-PCR) were used to detect the expression levels of Bax, Bcl-2 and Caspase-3.Results MCAO/R could induce the neuron injury in left cortex and hippocampus CAI area(P>0.05). And CZTLC significantly decreased the neurobehavioral scores and infarct area, and improved the oxidation state of brain tissues.In additional, CZTLC significantly increased the levels of anti-inflammatory cytokines(IL-4 and IL-10),and decreased the expression levels of pro-inflammatory cytokines(TNF-α and IL-1β).Furthermore, CZTLC increased Bcl-2 levels and decreased the expression levels of Bax and caspase-3 mRNA and protein(P<0.05).Conclusion CZTLC has definite protective effects on cerebral ischemia-reperfusion injury in mice, and its action mechanism is related to improving inflammatory response and inhibiting cell apoptosis.更多还原