夹脊电针不同时间点干预对ALS-SOD1^(G93A)小鼠腰髓TLR4/NF-κB信号通路的影响  

Effect of Jiaji EA at Different Time Points on TLR4/NF-κB Signaling Pathway in Lumbar Medulla of Mice with ALS-SOD1^(G93A)

作　　者：王仕林 孙远征[2] 于天洋 孙妍 WANG Shilin;SUN Yuanzheng;YU Tianyang;SUN Yan(Heilongjiang University of Chinese Medicine, Harbin 150040, China;The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin 150001, China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]黑龙江中医药大学附属第二医院,黑龙江哈尔滨150001

出　　处：《针灸临床杂志》2022年第6期56-62,共7页Journal of Clinical Acupuncture and Moxibustion

基　　金：国家中医药管理局全国名老中医药专家传承工作室建设项目,编号:国中医药人教发[2014]20号;黑龙江省自然科学基金联合引导项目,编号:LH2019H103;黑龙江中医药大学科研基金面上项目,编号:2019MS21。

摘　　要：目的:观察夹脊电针不同时间点干预对ALS-SOD1^(G93A)小鼠发病期、运动功能及TLR4/NF-κB信号通路相关因子TLR4、NF-κB和IL-1β的影响,探究夹脊电针治疗ALS的最佳干预时间及机制。方法:将36只ALS-SOD1^(G93A)小鼠随机分为模型组、60 d夹脊电针组和90 d夹脊电针组,12只/组;选取同窝经PCR鉴定的12只ALS-SOD1^(G93A)阴性鼠作为野生组。60 d夹脊电针组和90 d夹脊电针组分别在60日龄和90日龄时将小鼠固定在无菌操作台,针刺双侧L_(1~2)、L_(5~6)夹脊穴,针刺深度2~3 mm,连接电针2 Hz、1 mA,20 min/次,2次/周,共4周;模型组和野生组在小鼠60日龄时,将小鼠固定在无菌操作台,20 min/次,2次/周,共4周,不予治疗。采用悬尾实验判定小鼠发病日龄;转棒式疲劳仪评价小鼠肢体运动功能变化;各组小鼠均于120日龄取材,HE染色观察各组小鼠腰髓前角运动神经元形态;尼氏染色观察各组小鼠腰髓前角运动神经元数量;Western Blot法检测小鼠腰髓TLR4、NF-κB及IL-1β相对表达量。结果:经治疗后,模型组与90 d夹脊电针组发病时间比较,差异无统计学意义(P>0.05),60 d夹脊电针组发病时间明显推迟(P<0.01);与野生组比较,其他各组小鼠转棒时间均降低,差异具有统计学意义(P<0.05),其中60 d夹脊电针组较模型组、90 d夹脊电针组降低时间延后,差异具有统计学意义(P<0.05);与野生组比较,各组运动神经元数量均减少,差异具有统计学意义(P<0.05),其中60 d夹脊电针组运动神经元数量较模型组、90 d夹脊电针组保留较多,差异具有统计学意义(P<0.05);与野生组比较,各组TLR4、NF-κB及IL-1β表达均增加,差异具有统计学意义(P<0.05),其中60 d夹脊电针组较模型组、90 d夹脊电针组表达降低,差异具有统计学意义(P<0.05)。结论:夹脊电针可以延缓ALS-SOD1^(G93A)小鼠的发病时间,改善小鼠肢体活动功能,且60日龄时的早期干预疗效优于90日龄时干预,其机制可能与抑�Objective:To observe the effects of Jiaji electro-acupuncture(EA)at different time points on onset stage,motor function,TLR4/NF-κB signaling pathway related factors of TLR4,NF-κB and IL-1βin amyotrophic lateral sclerosis(ALS)-SOD1^(G93A) mice,and to explore the best intervention time and the mechanism of Jiaji EA in treatment of ALS.Methods:36 ALS-SOD1^(G93A) mice were randomly divided into the model group,the 60-day Jiaji EA group and the 90-day Jiaji EA group,with 12 mice in each group.12 ALS-SOD1^(G93A) negative mice identified by PCR were selected into the wild group.At 60 days and 90 days of age,the Jiaji EA groups were treated with electro-needling L_(1~2) and L_(5~6) Jiaji points on both sides(needling depth of 2 mm to 3 mm,2 Hz,1 mA,20 min/time,twice/week for 4 weeks).At 60 days and 90 days of age,the mice in the model group and the wild group were fixed on a sterile workbench for 20 min/time and twice/week for 4 weeks without treatment.The age of onset was determined by tail suspension experiment.The change of locomotor function of mice limb was evaluated by rotary rod fatigue instrument.The samples of rats were collected at 120 days of age,and the morphology of motonneurons in lumbar premedullary horn was observed by HE staining.The number of motonneurons in the lumbar premedullary horn of each group was observed by Nissner staining.The relative expressions of TLR4,NF-κB and IL-1βwere detected by Western Blot.Results:There was no statistical difference in the onset time between the model group and the 90-day Jiaji EA group after the treatment(P>0.05),whereas the onset time of the 60-day Jiaji EA group was significantly delayed(P<0.01).Compared with that in the wild group,the time of rod rotation in the other groups was decreased(P<0.05),and the reduction time of the 60-day Jiaji EA group was delayed compared with that in the model group and the 90-day Jiaji EA group(P<0.05).Compared with that in the wild group,the number of motor neurons in all other groups was decreased(P<0.05),and the number of m

关 键 词：肌萎缩侧索硬化症 TLR4/NF-κB信号通路 治疗时间 夹脊 电针 运动神经元 蛋白印迹法 

分 类 号：R246.6[医药卫生—针灸推拿学]