夹脊穴针刺联合低频电刺激对脊髓损伤大鼠自噬相关因子及膀胱功能的影响  

Effect of Acupuncture at Jiaji Acupoints Combined with Low-Frequency Electrical Stimulation on Autophagy-Related Factors and Bladder Function in Rats with Spinal Cord Injury

作　　者：李格格 董宝强[1] 焦政 马铁明[1] 陈怡然 王树东[1] 林星星[1] 王列[1] 田辉[1] 刘玉丽[1] 张淑慧[1] 王成龙 胡哲 马俊杰 LI Gege;DONG Baoqiang;JIAO Zheng;MA Tieming;CHEN Yiran;WANG Shudong;LIN Xingxing;WANG Lie;TIAN Hui;LIU Yuli;ZHANG Shuhui;WANG Chenglong;HU Zhe;MA Junjie(College of Acupuncture and Tuina,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)

机构地区：[1]辽宁中医药大学针灸推拿学院,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《中华中医药学刊》2024年第11期75-80,I0021-I0025,共11页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(82305391);辽宁省教育厅青年项目(JYTQN2023461);辽宁中医药大学自然科学类项目(2021 LZY017,2021 LZY030)。

摘　　要：目的观察夹脊穴针刺联合低频电刺激对T7脊髓打击伤大鼠自噬相关因子表达的影响,并探讨其影响膀胱功能的机制。方法50只雌性SD大鼠随机分为空白组(K)、假手术组(J)、模型组(M)、低频组(D)和低频联合针刺组(DZ),每组10只。D组Allen’s打击伤后接受50 Hz低频电刺激治疗,1次/d,每次20 min,连续治疗7 d;DZ组在每日低频电刺激治疗后10 min,直刺夹脊穴。采用尿动力学法检测治疗前后膀胱最大容量的差异;透射电镜观察脊髓组织线粒体自噬情况;免疫荧光法检测微管相关蛋白1 A/1 B轻链3 B(microtubule-associated proteins 1 A/1 B light chain 3 B,LC3 B)和分选体蛋白1(sequestosome 1,p62)的表达;免疫组化法检测神经生长因子(nerve growth factor,NGF)和酪氨酸蛋白激酶受体A(tysrosine kinase A,TrKA)的表达;蛋白质印迹法检测脊髓组织磷脂酰肌醇3-激酶(phosphatidylinositol-3-Kinase,PI3 K)信号通路相关蛋白,LC3 B和p62的表达。结果K组和J组脊髓组织神经细胞核结构完整、线粒体形态正常;M组核膜不清楚、线粒体空泡伴有自噬;D组核膜形态无差异,空泡化线粒体伴有自噬表现;而与M组相比,DZ组核膜形态和线粒体空泡方面无差异,但无明显自噬。与K组相比,J组治疗前最大膀胱容量、治疗后最大膀胱容量、治疗前后膀胱容量差值、NGF、TrKA、PI3 K、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-AKT)/蛋白激酶B(Protein Kinase B,AKT)、磷酸化哺乳动物雷帕霉素靶蛋白(phosphorylated mammalian target of rapamy-cin,p-mTOR)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)、LC3 B和p62表达没有统计学意义(P>0.05);与J组相比,M组治疗前最大膀胱容量、治疗后最大膀胱容量、治疗前后膀胱容量差值,LC3 B表达明显升高(P<0.01,P<0.05),NGF、TrKA、PI3 K、P-AKT/AKT、p-mTOR/mTOR、p62表达显著下降(P<0.01);与M组相比,D组治疗后最大膀胱容量与LC3 B表达显著下降(P<0.01)、治Objective To observe the effect of acupuncture at Jiaji acupoints combined with low-frequency electrical stimulation on the expressions of autophagy related factors in T7 spinal cord injury rats,and to explore the mechanism of its effect on bladder function.Methods Fifty female SD rats were randomly divided into blank group(K),sham operation group(J),model group(M),low frequency group(D)and low frequency combined acupuncture group(DZ),with 10 rats in each group.Group D was treated with 50 Hz low-frequency electrical stimulation once a day,20 min for each time,lasting for 7 days after Allen's blow injury.In group DZ,10 minutes after daily low-frequency electrical stimulation,Jiaji acupoints were directly needled.The difference of maximum bladder volume before and after treatment was detected by urodynamic method.The autophagy of mitochondria in spinal cord was observed by transmission electron microscope.The expressions of LC3 B and p62 were detected by immunofluorescence assay.The expressions of nerve growth factor(NGF)and tysrosine kinase A(TrKA)were detected by immunohistochemistry.The expressions of phosphatidylinositol 3-kinase(PI3 K)signaling pathway related proteins,LC3 B and p62 were detected by Western blotting.Results In group K and group J,the nucleus structure of the spinal cord was intact and the mitochondrial morphology was normal.In group M,the nuclear membrane was unclear and mitochondria vacuole was accompanied by autophagy.There was no difference in the morphology of nuclear membrane in group D,and vacuolated mitochondria were associated with autophagy.Compared with those in group M,there was no difference in nuclear membrane morphology or mitochondrial vacuole in group DZ,but there was no obvious autophagy.Compared with those in group K,there was no significant difference in the maximum bladder volume before treatment,maximum bladder volume after treatment,bladder volume difference before and after treatment,the levels of NGF,TrKA,PI3 K,phosphorylated protein kinase B(AKT)protein(p-AKT)/AKT,phos

关 键 词：夹脊穴 针刺 低频电刺激 自噬 磷脂酰肌醇3-激酶 

分 类 号：R245.97[医药卫生—针灸推拿学]