电针“夹脊”对腰椎间盘退变兔退变椎间盘髓核细胞衰老的影响  

Effect of electroacupuncture at“Jiaji”(EX-B2)on senescence of nucleus pulposus cells of degenerated lumbar intervertebral disc in rabbits

作　　者：张艳琳 邹璟[2] 汪敏 王昆秀 黄国付 ZHANG Yan-lin;ZOU Jing;WANG Min;WANG Kun-xiu;HUANG Guo-fu(College of Acupuncture and Orthopedics,Hubei University of Chinese Medicine,Wuhan 430061,China;Wuhan First Hospital,Wuhan 430030;Longgang District Maternity&Child Healthcare Hospital of Shenzhen/Longgang Maternity and Child Clinical School,Medical College,Shantou University,Shenzhen 518172,Guangdong Province)

机构地区：[1]湖北中医药大学针灸骨伤学院,武汉430061 [2]武汉市第一医院,武汉430030 [3]深圳市龙岗区妇幼保健院/汕头大学医学院龙岗妇幼临床学院,广东深圳518172

出　　处：《针刺研究》2023年第10期1001-1008,1047,共9页Acupuncture Research

基　　金：国家自然科学基金项目(No.82074517,81774392)。

摘　　要：目的:观察电针“夹脊”对腰椎间盘退变兔退变椎间盘髓核细胞及纤维环组织小窝蛋白1(Cav-1)阳性表达、髓核细胞端粒酶活性、相对端粒长度及不同细胞周期比例的影响,探讨电针“夹脊”延缓腰椎间盘退变髓核细胞衰老的作用机制。方法:新西兰大白兔按照随机数字表法分为正常组、假手术组、模型组、电针组、针刺组,每组5只。造模兔在腰椎(L)4、L5两椎体间穿入克氏针,给予持续轴向加压造模器加压28 d复制腰椎间盘退变模型。电针组取L4、L5双侧“夹脊”进行电针治疗,针刺组针刺L4、L5双侧“夹脊”,均治疗20 min,连续6 d为1个疗程,疗程间隔1 d,共治疗4个疗程。观察各组兔一般情况,每周测量其体质量变化,并分别于造模前、造模术后、造模结束后及治疗后进行运动功能评分,免疫组织化学法检测各组兔髓核细胞及纤维环组织Cav-1阳性细胞数,PCR-ELISA法检测髓核细胞端粒酶活性,荧光定量PCR法检测髓核细胞相对端粒长度,流式细胞仪检测髓核细胞周期比例。结果:与假手术组比较,模型组大白兔造模后每周体质量增长减缓(P<0.01),运动功能评分降低(P<0.01),髓核细胞及纤维环组织Cav-1阳性细胞数增多(P<0.01),髓核细胞端粒酶活性减弱(P<0.01),相对端粒长度缩短(P<0.01),G0/G1期细胞比例升高(P<0.01),G2/M期细胞比例降低(P<0.01)。与模型组比较,电针组大白兔治疗后每周体质量增长加快(P<0.01),运动功能评分升高(P<0.01),髓核细胞及纤维环组织Cav-1阳性细胞数减少(P<0.01),髓核细胞端粒酶活性增强(P<0.01),相对端粒长度增长(P<0.01),G0/G1期细胞比例降低(P<0.01),G2/M期细胞比例升高(P<0.01)。结论:电针“夹脊”有利于椎间盘退变模型兔的良性生长和运动功能的恢复,并可能通过下调髓核细胞及纤维环组织Cav-1阳性细胞数、增强髓核细胞端粒酶活性、延长髓核细胞相对端粒长度和改善髓核细Objective To observe the effect of electroacupuncture(EA)at“Jiaji”(EX-B2)on body mass,mo⁃tor function,expression of caveolin-1(Cav-1)in nucleus pulposus cells and annulus fibrosus tissue,telomerase activi-ty,relative telomere length and different cell cycle ratio of nucleus pulposus cells in rabbits with intervertebral disc de⁃generation(IVDD),so as to investigate its mechanism underlying delaying senescence of the degenerated lumbar inter⁃vertebral disc nucleus pulposus cells.Methods Twenty-five male New Zealand rabbits with mature bones were di⁃vided into control,sham operation,model,EA,and acupuncture groups,with 5 rabbits in each group.The IVDD model was established by inserting kirschner wires to the vertebral bone surface between the lumbar(L)4 and L5 verte⁃brae,followed by applying continuous axial pressure for 28 d.EA(2 Hz/15 Hz,1—2 mA)or acupuncture(only inser⁃tion of acupuncture needles into bilateral EX-B2,but without electrical stimulation)was applied to bilateral EX-B2 for 20 min,once daily,6 times a week for 4 weeks.The hindlimb locomotor function(locomotor score)was assessed by using Faden’s and colleagues’methods.The general conditions of rabbits in each group were observed,and their body weight changes were measured every week.Nucleus pulposus cells were isolated using enzyme digestion method.After the treatment,the Cav-1 positive cell counts in nucleus pulposus cells and annulus fibrosus tissues were detected by immunohistochemistry,and the telomerase activity of nucleus pulposus cells was detected by PCR-ELISA.The relative telomere length of nucleus pulposus cells was measured by real-time quantitative polymerase chain reac⁃tion(real-time qPCR),and the cell cycle of nucleus pulposus was detected by flow cytometry.Results Compared with the sham operation group,the body mass from 4 to 11 week,locomotor score at 4,7 and 11 week,telomerase activity,relative telomere length and the proportion of cells in G2/M phase of nucleus pulposus cells were significantly decreased(P<0.01

关 键 词：椎间盘退变 电针 细胞衰老 小窝蛋白1 端粒 细胞周期 

分 类 号：R245.31[医药卫生—针灸推拿学]