中国中药杂志

2020, v.45(04) 932-936

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β-石竹烯通过激活自噬减轻小鼠脑缺血/再灌注损伤的研究
β-caryophyllene alleviates cerebral ischemia/ reperfusion injury in mice by activating autophagy

饶江燕;王倩;王钰淳;向菲;田晓翠;刘道航;董志;
RAO Jiang-yan;WANG Qian;WANG Yu-chun;XIANG Fei;TIAN Xiao-cui;LIU Dao-hang;DONG Zhi;Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology;Department of Pharmacy, School of Chongqing Medical University;

摘要(Abstract):

脑缺血再灌注(ischemia/reperfusion,I/R)损伤是急性缺血性脑卒中的重要诱因,而在脑缺血再灌注过程中通过调节自噬及时清除受损蛋白质和细胞器,对减轻脑损伤起着重要作用。该文为了探讨β-石竹烯(beta-caryophyllene,BCP)是否可以通过调节自噬对I/R损伤小鼠起到神经保护作用,将C57BL/6雄性小鼠随机分为假手术组、模型组、给药组,各组小鼠术前连续灌胃给药5 d后,采用线栓法建立小鼠右侧大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型。术后24 h对脑梗死体积和神经功能进行评价;HE染色法观察缺血区皮层组织病理改变;Western blot法检测自噬相关蛋白beclin1,p62,LC3B和凋亡相关蛋白Bcl-2的表达情况;免疫荧光法观察LC3B在缺血区皮层组织中的表达;透射电镜观察缺血区皮层组织细胞自噬现象。结果显示,与模型组相比,BCP预处理可明显减轻小鼠的神经功能缺损,降低脑梗死体积百分率,减少缺血区脑组织细胞的死亡,上调beclin1,LC3B,Bcl-2蛋白的表达,下调p62蛋白的表达,明显增加缺血区皮层组织自噬体的数量,最终确定BCP可以通过激活自噬对I/R损伤小鼠起到较好的神经保护作用。
Cerebral ischemia-reperfusion(I/R) injury is an important cause of acute ischemic stroke. Timely elimination of damaged proteins and organelles by regulating autophagy during cerebral ischemia-reperfusion plays an important role in relieving brain damage. In order to investigate whether β-caryophyllene(BCP) could protect neurons from cerebral I/R injury by regulating auto-phagy, C57 BL/6 J male mice were randomly divided into sham operation group, model group, and drug-administered group. After intra-gastric administration was given for 5 days, the middle cerebral artery occlusion(MCAO) model was established by suture method. Twenty four hours after surgery, the infarct volume and neurological function were assessed; the pathological changes of cortical tissue were observed by HE staining; Western blot was used to detect the expression of autophagy-related proteins beclin1, p62, LC3 B and apoptosis-related protein Bcl-2; immunofluorescence was used to observe the expression of LC3 B in the ischemic cortex. The autophagy of cortical tissue in the ischemic area was observed by transmission electron microscopy. The experimental results showed that as compared with the model group, the BCP pretreatment significantly reduced the neurological deficit, decreased the percentage of cerebral infarction volume, reduced the death of brain tissue cells in the ischemic area, up-regulated the expression of beclin1, LC3 B and Bcl-2 protein, down-regulated p62 protein expression, and significantly increased the number of autophagosomes in the cortical tissue of the ischemic area. It was finally determined that BCP could protect neurons from cerebral ischemia-reperfusion injury by activating autophagy.

关键词(KeyWords): 脑缺血再灌注损伤;β-石竹烯;自噬;神经保护;beclin1
cerebral ischemia-reperfusion injury;β-caryophyllene;autophagy;neuroprotection;beclin1

Abstract:

Keywords:

基金项目(Foundation): 重庆市基础研究与前言探索一般课题(stc2018jcyjAX0378)

作者(Author): 饶江燕;王倩;王钰淳;向菲;田晓翠;刘道航;董志;
RAO Jiang-yan;WANG Qian;WANG Yu-chun;XIANG Fei;TIAN Xiao-cui;LIU Dao-hang;DONG Zhi;Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology;Department of Pharmacy, School of Chongqing Medical University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20191112.402

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