摘要
目的从自噬探讨醒神通督针刺抗脑缺血再灌注损伤的作用机制。方法雄性SD大鼠随机分为假手术组、脑缺血再灌注模型组(简称模型组)和脑缺血再灌注+醒神通督针刺组(简称针刺组)。采用大脑中动脉线栓法构建脑缺血再灌注损伤模型,针刺组大鼠造模前5 d开始干预,取穴“百会”“四神聪”“足三里”,每日针刺1次,直至再灌注24 h。另外2组大鼠只抓取不针刺。再灌注24 h后采用氯化三苯基四氮唑(triphenyltetrazolium chloride,TTC)染色观察脑梗死面积,透射电镜观察神经元及自噬的亚显微结构,Western blot、免疫荧光检测缺血皮层微管相关蛋白1轻链3(microtubule-associated protein light chain 3B,LC3)、肌球蛋白样BCL2结合蛋白(coiled coil moesin like BCL2 interacting protein,Beclin-1)、自噬相关蛋白(autophagy related protein,Atg)12、蛋白激酶B(protein kinase B,PKB,又称Akt)与磷酸化Akt(phosphorylated Akt,p-Akt)的表达。结果与假手术组比较,模型组大鼠大脑中动脉供血区梗死面积明显增加(P<0.05),且缺血皮层神经元结构严重破坏,自噬明显激活,自噬蛋白Beclin-1、Atg12表达及LC3Ⅱ/I比值显著升高(P<0.05),p-Akt表达下降(P<0.05)。与模型组比较,针刺组大鼠脑梗死面积减少(P<0.05),透射电镜显示神经元结构较为正常,自噬体较少。此外,针刺组自噬蛋白Beclin-1、Atg12表达及LC3Ⅱ/I比值明显降低(P<0.05),p-Akt表达显著上调(P<0.05)。结论针刺具有抗脑缺血再灌注损伤作用,可能通过抑制自噬的活化实现。
Objective To investigate the effects of Xingshen Tongdu acupuncture on rats after cerebral ischemia and reperfusion based on autophagy.Methods Male SD rats were randomly divided into 3 groups:sham control(Sham),ischemia and reperfusion(IR),IR plus Xingshen Tongdu acupuncture(Acu).The model was induced by middle cerebral artery occlusion(MCAO)followed by reperfusion.The acupuncture intervention was started 5 days before modeling to the rats of the Acu group,and the acupoints“Baihui”“Sishencong”and“Zusanli”were selected for treatment,once a day until 24 h after reperfusion.Rats in the other 2 groups were only grabbed without acupuncture.After 24 h of reperfusion,cerebral infarct areas of rats were evaluated by triphenyltetrazolium chloride(TTC)staining in all groups.Ultrastructural morphologies of autophagy were observed by transmission electron microscope(TEM).Levels of microtubule-associated protein light chain 3B(LC3),coiled coil moesin like BCL2 interacting protein(Beclin-1),autophagy related protein(Atg)12,protein kinase B(Akt)and p-Akt were detected by Western blot and immunofluorescent staining.Results Compared with the Sham group,cerebral infarct areas of MCA territory were increased(P<0.05),and TEM indicated that neuronal autophagy was significantly activated in the rats of the IR group.Moreover,Beclin-1,Atg12 expression levels,and the ratio of LC3Ⅱ/I were significantly up-regulated(P<0.05),while the p-Akt level was decreased(P<0.05)in the rats of the IR group.Compared with the IR group,the cerebral infarct area decreased(P<0.05),while TEM revealed a relatively normal neuronal structure with fewer autophagosomes in the rats of Acu group.Furthermore,the expression of Beclin-1 and Atg12 as well as LC3Ⅱ/I ratio decreased obviously(P<0.05),while the expression of p-Akt protein was significantly up-regulated(P<0.05)in the rats of Acu group.Conclusion The protective effects of acupuncture on cerebral ischemia and reperfusion injury may be related to inhibiting autophagy.
作者
兰瑞
邹旭欢
马云枝
付雪琴
王漫漫
王玮玮
张勇
LAN Rui;ZOU Xuhuan;MA Yunzhi;FU Xueqin;WANG Manman;WANG Weiwei;ZHANG Yong(The First Af liated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;Henan University of Chinese Medicine,Zhengzhou 450000,China;The Third Af liated Hospital of Zhengzhou University,Zhengzhou 450000,China)
出处
《中国中医基础医学杂志》
CAS
CSCD
2024年第4期633-638,共6页
JOURNAL OF BASIC CHINESE MEDICINE
基金
国家自然科学基金项目(81973618,81503422)
河南省自然科学基金项目(202300410399)。
关键词
醒神通督
针刺
脑缺血再灌注损伤
自噬
Xingshen Tongdu
Acupuncture
Cerebral ischemia and reperfusion injury
Autophagy
