电针对创伤性颅脑损伤大鼠损伤区皮层自噬相关蛋白表达的影响  

Effect of electroacupuncture on expression of corticocerebral autophagy-related proteins in rats with traumatic brain injury

作　　者：谷婷 吴涛 王瑞辉 杨欢 柯增辉 王东 陈星 杨挺 孟晓鹏 GU Ting;WU Tao;WANG Rui-hui;YANG Huan;KE Zeng-hui;WANG Dong;CHEN Xing;YANG Ting;MENG Xiao-peng(College of Acupuncture and Massage,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China;Medical Research Experimental Center,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China)

机构地区：[1]陕西中医药大学,针灸推拿学院,陕西咸阳712046 [2]陕西中医药大学,医学科研实验中心,陕西咸阳712046

出　　处：《针刺研究》2020年第7期524-528,547,共6页Acupuncture Research

基　　金：国家自然科学基金项目(No.81674088、81904310);陕西省中医药管理局课题(No.2019-zz-JC023);陕西省自然科学基金青年项目(No.2018JQ8070);陕西省教育厅专项科学研究计划(No.19JK0239);陕西省高校科协青年人才托举计划(No.20180308)。

摘　　要：目的:观察电针对创伤性颅脑损伤(TBI)大鼠大脑皮层自噬相关蛋白表达的影响,探讨电针治疗TBI的可能机制。方法:将雄性SD大鼠随机分为假手术组、模型组、电针Ⅰ组、电针Ⅱ组,每组10只。采用改良Feeney自由落体打击法制备TBI模型。电针Ⅰ组于造模后第7天开始电针"内关""足三里"联合针刺"水沟""百会",1次/d,连续7 d;电针Ⅱ组取穴及治疗方法同电针Ⅰ组,于造模后24 h开始干预,1次/d,连续14 d;假手术组只钻开颅骨,不造模也不进行任何干预。治疗结束后,取各组大鼠损伤区脑组织皮层用HE染色和尼氏染色法观察病理形态变化;Western blot法检测损伤区脑组织皮层AMP依赖的蛋白激酶(AMPK)、磷酸化AMPK(p-AMPK)、哺乳动物雷帕霉素靶蛋白(mTOR)、磷酸化mTOR(p-mTOR)、Ulk1、磷酸化Ulk1(p-Ulk1)蛋白表达情况。结果:与假手术组比较,模型组大鼠大脑损伤区皮层可见大量组织坏死,神经纤维排列散乱,细胞空泡样改变,核破碎、固缩,有增生的瘢痕组织;尼氏小体明显减少;创伤区皮层p-AMPK/AMPK上调(P<0.01),p-mTOR/mTOR、p-Ulk1/Ulk1均下调(P<0.01)。与模型组比较,两电针组大鼠大脑皮层损伤区病理改变减轻;尼氏小体数量增加;损伤区皮层p-AMPK/AMPK下调(P<0.01),p-mTOR/mTOR、p-Ulk1/Ulk1均上调(P<0.01)。与电针Ⅰ组比较,电针Ⅱ组损伤区皮层病理改善情况更明显;p-AMPK/AMPK下调(P<0.05),p-mTOR/mTOR、p-Ulk1/Ulk1均上调(P<0.05)。结论:电针可能是通过调节AMPK、mTOR、Ulk1自噬相关蛋白的活化状态来抑制创伤区皮层神经元自噬的过度活化,进而对TBI大鼠发挥脑保护作用,且电针早期干预效果更佳。Objective To observe the effect of electroacupuncture on the expression of phosphorylated adenosine 5’-monophosphate-activated protein kinase(p-AMPK),phosphorylated mammalian target of rapamycin(p-mTOR)and phosphorylated Ulk1(p-Ulk1)proteins in the cortex of traumatic brain injury(TBI)rats,so as to explore its mechanisms underlying treatment of TBI.Methods Male SD rats were randomly divided into sham operation(sham),model,electroacupunctureⅠ(EAⅠ),electroacupunctureⅡ(EAⅡ)groups(n=10 in each group).TBI model was established by using a free fall brain injury striking device after exposing the local cranial bone(to induce the left parietal cerebral contusion).Rats in EAⅠgroup were treated by electroacupuncture at"Neiguan"(SP6)and"Zusanli"(ST36)combined with acupuncture at"Shuigou"(GV26)and"Baihui"(GV20)on the 7thday after modeling,once a day for 7 consecutive days.Rats in EAⅡgroup received the treatments as those in EAⅠgroup on 24 h after modeling,once a day for 14 d.After the treatment,histopathological changes of the injured cerebral cortex were observed by HE staining and Nissl staining.Western blot was used to detect the expression of AMPK,p-AMPK,mTOR,p-mTOR,Ulk1,p-Ulk1 proteins in the injured cerebral cortex tissue.Results After modeling and compared with the sham group,a large number of tissue necrosis,scattered arrangement of nerve fibers,vacuolar changes of cells,nuclear fragmentation,consolidation and hyperplastic scar tissue were found in the brain trauma area of rats in the model group.Nissl corpuscles were obviously absent.The ratio of p-AMPK/AMPK was up-regulated in the cortex of the wound region(P<0.01),and the ratio of p-mTOR/mTOR,p-Ulk1/Ulk1 were down-regulated(P<0.01).Compared with the model group,the pathological changes in brain injury area of rats in both EA groups were alleviated,the number of Nissl corpuscles increased,the ratio of p-AMPK/AMPK was down-regulated in cortex of the injury area(P<0.01),and the ratios of p-mTOR/mTOR and p-Ulk1/Ulk1 were up-regulated(P<0.01).Compared w

关 键 词：创伤性颅脑损伤 电针 自噬 AMP依赖的蛋白激酶 哺乳动物雷帕霉素靶蛋白 Ulk1 

分 类 号：R245.9+7[医药卫生—针灸推拿学]