摘要
目的:探讨加味涤痰汤对脑缺血再灌注损伤大鼠脑细胞自噬相关蛋白表达的影响。方法:采用可逆性脑中动脉闭塞线栓法栓塞右侧大脑中动脉,构建脑缺血再灌注(CIR)损伤模型,随机分为假手术组、模型组、加味涤痰汤高、低剂量组(0. 768,0. 384 g·kg-1)和吡拉西坦组(0. 1 g·kg-1)。假手术组和模型组灌胃生理盐水,加味涤痰汤高、低剂量组灌胃加味涤痰汤,吡拉西坦组灌胃吡拉西坦片,灌胃10 m L·kg-1;给药7 d。给药24 h内,处死后行组织病理学检查,比较脑梗死体积、神经细胞凋亡和血清炎症因子水平,蛋白免疫印迹法(Western blot)测定脑组织中自噬相关蛋白微管相关蛋白轻链3(LC3)Ⅱ,Beclin1,B淋巴细胞瘤-2(Bcl-2)和p62的表达。结果:模型组脑组织梗死灶内细胞和血管坏死,神经元肿胀,间质水肿;加味涤痰汤高、低剂量组及吡拉西坦组脑组织少数神经细胞死亡,水肿减轻,神经细胞肿胀减轻。加味涤痰汤高、低剂量组及吡拉西坦组脑梗死体积和神经细胞凋亡低于模型组(P <0. 05),加味涤痰汤高、低剂量组及吡拉西坦组肿瘤坏死因子-α(TNF-α),白细胞介素-2(IL-2)和白细胞介素-8(IL-8)水平降低(P <0. 05)。与假手术组比较,模型组LC3Ⅱ,Bcl-2和Beclin1蛋白及mRNA表达明显升高,p62蛋白及mRNA表达明显下降;与模型组比较,加味涤痰汤高、低剂量组及吡拉西坦组LC3Ⅱ和Beclin1蛋白及mRNA表达均明显降低,p62蛋白及mRNA表达明显升高,差异均具有明显性(P <0. 05)。结论:加味涤痰汤可改善MCAO/R脑组织损伤和自噬,减轻炎症反应,调节自噬活性,可能与下调LC3Ⅱ,Beclin1表达,上调p62表达有关。
Objective:To investigate the effect of modified Ditantang on autophagy and relevant proteins in brain cells of rats with cerebral ischemia reperfusion injury.Method:The cerebral ischemic reperfusion(CIR)injury model in rats was built by reversible middle cerebral artery occlusion artery suture of middle cerebral embolism method,and randomly divided into sham group,model group,high-dose modified Ditantang group(H-dose),low-dose modified Ditantang group(L-dose,0.384 g·kg-1)and PLXT group(0.1 g·kg-1).Sham and model groups were given normal saline by gastric perfusion,H-dose and L-dose groups were given modified Ditantang,and the PLXT group were given Piracetam tablets,intragastric volume 10 m L·kg-1.The treatment lasted for 7 d.Within 24 hours after administration,the histopathological examination was performed,the volume of cerebral infarction,neuronal apoptosis and serum levels of inflammatory factors were compared,and the expressions of autophagy related microtuble-associated protein 1 light chain 3(LC3)Ⅱ,Beclin1,B-cell lymphoma-2(Bcl-2)and p62 in brain tissues were determined.Result:Cells and blood vessel necrosis,neuron swelling and interstitial edema were observed in model group,a few neurons died,edema was reduced,swelling of nerve cells was alleviated in H-dose,L-dose and PLXT groups.The volume of cerebral infarction and neuronal apoptosis in Hdose,L-dose and PLXT groups were lower than those in model group(P<0.05).The levels of tumor necrosis factor(TNF)-α,intedeukin(IL)-2 and IL-8 in H-dose,L-dose and PLXT groups were lower than those in model group(P<0.05).Compared with sham group,protein and mRNA expressions of LC3Ⅱ,Bcl-2 and Beclin1 in the ischemic brain tissue of model group were significantly increased,while protein and mRNA expressions of p62 in the ischemic brain tissue of model group were decreased significantly.Compared with the model group,protein and mRNA expressions of LC3Ⅱand Beclin1 in H-dose,L-dose and PLXT groups were significantly decreased,while protein and mRNA expressions of p62 were significantly increased,with significant differences(P<0.05).Conclusion:Modified Ditantang can improve brain injury and interfere with autophagy after MCAO/R,alleviate inflammation,and regulate autophagic activity,which may be related to the downregulation of expressions of LC3Ⅱ,Beclin1 and the up-regulation of expression of p62.
作者
陈维达
宋婷
赵鑫
张琼瑜
刘晓婷
陈泽涛
CHEN Wei-da;SONG Ting;ZHAO Xin;ZHANG Qiong-yu;LIU Xiao-ting;CHEN Ze-tao(Shandong University of Traditional Chinese Medicine(TCM),Ji'nan 250355,China;Hospital Affiliated to Shandong University of TCM,Ji'nan 250011,China;Weifang TCM Hospital,Weifang 261041,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第15期64-69,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
山东省中医药科技发展计划项目(2015-076)
关键词
加味涤痰汤
脑缺血再灌注损伤
自噬
机制
modified Ditantang
cerebral ischemia reperfusion injury
autophagy
mechanism
