目的: 探讨肉苁蓉总苷(GCs)对脑缺血再灌注损伤(CIRI)大鼠氧化应激损伤及神经细胞凋亡的影响及可能机制。方法: 模型大鼠采用线栓法构建大脑中动脉闭塞(MCAO)模型。造模成功后,药物组(GCs)大鼠给与GCs 50 mg/(kg·d)灌胃,连续14 d;假手术组(Sham)及模型组(Model)大鼠给与同等体积生理盐水灌胃。给药结束后采用Longa评分法评价其神经功能缺损情况;随后处死大鼠,进行TTC染色计算脑梗死面积百分比;ELISA法检测血清丙二醛(MDA)含量、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)的活性;蛋白免疫印记(Western blot)检测脑组织中核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)以及B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)的表达情况。结果: 与Sham组相比,Model组神经功能缺损评分升高,脑梗死面积百分比升高,MDA含量升高,SOD和GSH-Px活性降低,Nrf 2、HO-1及Bcl-2蛋白表达水平下降,Bax及Caspase-3蛋白表达水平升高;与Model组相比,GCs组大鼠神经功能缺损评分有所降低,脑梗死面积百分比降低,MDA含量降低,SOD和GSH-Px活性升高,Nrf 2、HO-1及Bcl-2蛋白表达水平升高,Bax及Caspase-3蛋白表达水平下降。结论: GCs可有效地减轻脑缺血再灌注损伤,其机制可能与激活Nrf 2/HO-1通路,进而降低氧化应激水平,抑制神经细胞的凋亡有关。
Objective: To investigate the effects of total glycosides of cistanche (GCs) on oxidative stress injury and neuronal apoptosis in rats with cerebral ischemia-reperfusion injury (CIRI) and its possible mechanism. Methods: The rat model of middle cerebral artery occlusion (MCAO) was made by thread embolism. After successful modeling, rats in the drug group (GCs) were given GCs 50 mg/(kg·d) for 14 days; rats in the blank group (Sham) and model group (Model) were gavaged with the same volume of normal saline. After the end of administration, Longa score was used to evaluate the neurological deficit. Subsequently, the rats were sacrificed, and the percentage of cerebral infarction area was calculated by TTC staining. The content of malondialdehyde (MDA), the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in serum were detected by ELISA. Western blot was used to detect the expression of nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), B-cell lymphoma/leukemia-2 (Bcl-2 ), Bcl-2-related X protein (Bax) and cysteine aspartic protease-3 (Caspase-3) in brain tissue. Results: Compared with the Sham group, the neurological deficit score, the percentage of cerebral infarction area, MDA content, Bax and Caspase-3 protein expression levels in the Model group were increased, while SOD and GSH-Px activities,Bcl-2,Nrf 2,and HO-1 protein expression levels in the Model group were decreased; compared with the Model group, the neurological deficit score, the percentage of cerebral infarction area, MDA content, Bax and Caspase-3 protein expression levels in GCs group were decreased, while SOD and GSH-Px activities, Bcl-2, Nrf 2, and HO-1 protein expression levels in GCs group were increased. Conclusion: GCs can effectively reduce cerebral ischemia-reperfusion injury, which may be related to activating Nrf 2/HO-1 pathway, reducing oxidative stress levels, and inhibiting apoptosis of nerve cells.
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