氯喹对大鼠脑缺血再灌注自噬相关蛋白的影响

发布时间：2019-05-27 10:02

【摘要】：目的探讨氯喹对大鼠脑缺血再灌注自噬相关蛋白的影响及其机制。方法取75只大鼠随机分为假手术组(Sham),模型组(Model),氯喹(CQ)20、40、60 mg/kg组,每组15只。应用线栓法使右脑中动脉栓塞(MCAO),栓塞2 h。3个药物组于栓塞后立即腹腔注射CQ 20、40、60 mg/kg,其余各组注射2 ml 0.9%氯化钠。再灌注24 h后,采用神经功能缺损评分、TTC染色判断模型是否制作成功,采用免疫荧光和免疫印迹法检测大鼠脑自噬相关蛋白的表达。结果 (1)Sham组,Model组,CQ 20、40、60 mg/kg组神经功能缺损评分,分别为0、2.67±0.49、2.93±0.26、2.40±0.51及1.93±0.70。通过TTC染色与Model组比较梗死面积,CQ 20 mg/kg组增大,CQ 40 mg/kg组无明显变化,CQ 60 mg/kg组减小。(2)免疫荧光和免疫印迹结果显示各组均有蛋白(Atg5、Atg7、Beclin1、Atg12)表达,与Model组比较,其中Sham组仅有极少量表达(P0.05),CQ 20 mg/kg组表达量无明显差异,CQ 60 mg/kg组的表达量明显减少(P0.05)。结论氯喹对大鼠脑缺血再灌注损伤有双重作用,低剂量20 mg/kg加重损伤,高剂量60 mg/kg减轻损伤。其机制可能与某些脑自噬相关蛋白表达的量有关。
[Abstract]:Objective to investigate the effect of chloroquine on autophagy related proteins after cerebral ischemia-reperfusion in rats and its mechanism. Methods 75 rats were randomly divided into (Sham), model group (Model), chloroquine (CQ) 20, 40 mg/kg group and 60 mg/kg group with 15 rats in each group. The right middle cerebral artery was embolized with (MCAO), for 2 hours. CQ 20 was injected intraperitoneally in 3 drug groups immediately after embolism, 40, and 2 ml 0.9% sodium chloride was injected into the other groups at 60 mg/kg,. After 24 hours of reperfusion, the neurological deficit score and TTC staining were used to determine whether the model was successful or not. Immunofluorescence and immunoblotting were used to detect the expression of autophagy related proteins in the brain of rats. Results (1) the neurological deficit scores of Sham group, Model group, CQ 20, 40, 60 mg/kg group were 0, 2.67 卤0.49, 2.93 卤0.26, 2.40 卤0.51 and 1.93 卤0.70, respectively. Compared with Model group, the infarction area of CQ 20 mg/kg group increased by TTC staining, but there was no significant change in CQ 40 mg/kg group, but decreased in CQ 60 mg/kg group. (2) the results of immunofluorescence and immunoblotting showed that there was protein (Atg5,Atg7,Beclin1,) in all groups. Compared with Model group, there was only a small amount of expression in Sham group (P 0.05), CQ 20 mg/kg group), but the expression level of CQ 60 mg/kg group was significantly decreased (P 0.05). Conclusion chloroquine has a dual effect on cerebral ischemia-reperfusion injury in rats. Low dose 20 mg/kg aggravates the injury, and high dose 60 mg/kg alleviates the injury. The mechanism may be related to the expression of some brain autophagy related proteins.
【作者单位】：锦州医科大学细胞生物学教研室;锦州医科大学生命科学院;锦州医科大学附属第一医院口腔科;锦州医科大学解剖学教研室;锦州医科大学基础医学实验教学中心;
【基金】：国家自然科学基金(81202783) 省科技厅计划项目(2015020696,20170540378)
【分类号】：R743.3

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