针刺通过PI3K-Ⅲ/Beclin-1通路抑制脑缺血-再灌注损伤大鼠的细胞凋亡  

Acupuncture inhibits cell apoptosis via PI3K-I/Beclin-1 pathway in rats with cerebral ischemia-reperfusion injury

作　　者：江姗姗 吕倩忆 唐红 汪红娟 王瑶 谢灿明 陈楚淘[1] 田浩梅[1] JIANG Shanshan;LYU Qianyi;TANG Hong;WANG Hongjuan;WANG Yao;XIE Canming;CHEN Chutao;TIAN Haomei(School of Acupuncture-moxibustion,Tuina,and Rehabilitation,Hunan University of Chinese Medicine,Changsha 410208;Chengdu Second People's Hospital,Chengdu 610021,China)

机构地区：[1]湖南中医药大学针灸推拿与康复学院,长沙410208 [2]成都市第二人民医院,成都610021

出　　处：《神经解剖学杂志》2023年第2期179-186,共8页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81874508,82274662);湖南省自然科学基金(2020JJ4065,2021JJ30490);湖南省研究生科研创新课题(CX20220799);长沙市科技局自然科学基金(kq2014094);湖南中医药大学研究生创新课题(2021CX39,2022CX97)。

摘　　要：目的:利用大脑中动脉闭塞/再灌注(MCAO/R)法制备大鼠脑缺血再灌注损伤(CIRI)模型,研究针刺对CIRI模型大鼠脑组织中Ⅲ型磷脂酰肌醇-3-羟激酶(PI3K-Ⅲ)/Beclin-1通路的影响。方法:利用MCAO/R方法制备SD大鼠CIRI模型,分别给予针刺和(或)自噬抑制剂3-甲基腺嘌呤(3-MA)进行治疗。利用采用Garcia评分法检测大鼠神经功能,TTC染色法检测大鼠脑梗死面积,Western Blot法检测海马组织PI3K-Ⅲ、Beclin-1和B淋巴细胞瘤-2基因(Bcl-2)蛋白的表达水平,TUNEL法检测细胞凋亡。结果:MCAO/R手术后大鼠神经功能评分降低(P<0.01),TTC染色观察到脑梗死,同时海马区PI3K-Ⅲ、Beclin-1和Bcl-2表达显著下调(P<0.01),脑皮质细胞凋亡增多(P<0.01)。针刺治疗不但提高了大鼠神经功能评分(P<0.01),并且减少了脑梗死面积(P<0.01);同时显著上调了海马组织中PI3K-Ⅲ、Beclin-1和Bcl-2表达(P<0.01),并减少了脑细胞凋亡(P<0.01)。3-MA阻断了针刺的治疗作用,而且使脑损伤更加严重。结论:针刺可改善CIRI大鼠神经功能并减轻神经损伤,其机制可能与激活PI3K-Ⅲ/Beclin-1通路增加细胞自噬并减少细胞凋亡有关。Objective:To establish cerebral ischemia reperfusion injury(CIRI)rat model by middle cerebral artery occlusion/reperfusion(MCAO/R)method and investigate the effect of acupuncture on class Ⅲ phosphatidylinositol-3-hydroxykinase(PI3K-Ⅲ)/Beclin-1 pathway in the brain of CIRI model rats.Methods:CIRI model of SD rats was prepared by MCAO/R method and treated with acupuncture and/or autophagy inhibitor 3-methyladenine(3-MA).Garcia score was used to detect neurological function in rats.The cerebral infarct area of rats was observed by TTC staining.The expression levels of PI3K-Ⅲ,Beclin-1,and Bcl-2 in hippocampal tissues were determined by Western Blot.And the apoptosis rate of neuronal cells was detected by TUNEL syaining.Results:Neurologic scores of rats were decreased after MCAO/R operation(P<0.01).TTC staining showed cerebral infarction.The expressions of PI3K-Ⅲ,Beclin-1 and Bcl-2 in the hippocampus were significantly down-regulated(P<0.01).In addition,the apoptosis of brain cell increased(P<0.01).Acupuncture treatment not only improved the neurological function scores of rats(P<0.01),but also reduced infarct size.What's more,Acupuncture treatment significantly up-regulated the expressions of PI3KII,Beclin-1 and Bcl-2 in hippocampus(P<0.01)and significantly reduced the apoptosis of brain cells(P<0.01).3-MA blocked the therapeutic effect of acupuncture and made the brain injury more serious.Conclusion:Acupuncture can improve neurological function and reduce brain injury in CIRI rats,and the mechanism may be related to the activation of PI3K-Ⅲ/Beclin-1 pathway to increase autophagy and reduce apoptosis.

关 键 词：针刺 脑缺血再灌注损伤 Ⅲ型磷脂酰肌醇-3-羟激酶/Beclin-1通路 细胞凋亡 大鼠 

分 类 号：R245[医药卫生—针灸推拿学]