推拿按揉法对慢性坐骨神经挤压伤大鼠脊髓背角星形胶质细胞和神经元的影响  

Effects of Tuina on Astrocytes and Neurons in Spinal Dorsal Horn of Rats with Chronic Sciatic Nerve Crush Injury

作　　者：张幻真 林志刚 陈乐春 陈进城 江煜 蒋晶晶 陈水金 ZHANG Huan-zhen;LIN Zhi-gang;CHEN Le-chun;CHEN Jin-cheng;JIANG Yu;JIANG Jing-jing;CHEN Shui-jin(Fujian University of Chinese Medicine Subsidiary Rehabilitation Hospital,Fuzhou,Fujian 350003;Fujian Provincial Key Laboratory of Rehabilitation Technology,Fuzhou,Fujian 350003)

机构地区：[1]福建中医药大学附属康复医院,福建福州350003 [2]福建省康复技术重点实验室,福建福州350003

出　　处：《中医康复》2024年第4期1-5,共5页Traditional Chinese Medicine Rehabilitation

基　　金：国家自然科学基金(82105039);福建省自然科学基金(2022J01881);福建省卫健委课题(2020CXA052);福建省教育厅课题(JAT200224)。

摘　　要：目的:观察推拿按揉法对慢性坐骨神经挤压伤(CCI)大鼠星形胶质细胞和神经元的影响,探讨其治疗神经病理性疼痛的潜在作用机制。方法:将入组的40只SD大鼠随机平均分成5组(每组8只),分别为空白组、模型组、推拿组、星形胶质细胞抑制剂组和星形胶质细胞抑制剂+推拿组。除空白组外的组别均建立CCI模型,造模后第4天起,推拿组接受按揉法干预,10分钟/次/天,星形胶质细胞抑制剂组鞘内注射氟代柠檬酸(fluorocitrate,FCA),10ul/只/天,星形胶质细胞抑制剂+推拿组进行鞘内注射FCA和按揉委中穴联合干预,连续14天。观察各组大鼠各时间点的机械痛阈值情况,使用免疫荧光检测脊髓背角星形胶质细胞标志物(glialfibrillaryacidicprotein,GFAP)蛋白的表达,尼式染色法观测脊髓背角内神经元的形态、排列和数量的变化,并分析GFAP表达水平与神经元存活数量的相关性。结果:相较空白组,模型组机械痛阈值显著下降(P<0.01);推拿干预后,推拿组阈值较模型组逐渐上升,累计治疗14天后具有显著差异(P<0.01),注射FCA第7天起,星形胶质细胞抑制剂组和星形胶质细胞抑制剂+推拿组疼痛阈值明显上升(P<0.01)。相较空白组,模型组脊髓背角GFAP表达显著升高(P<0.01),神经元形态不规则、排列松散,数量减少(P<0.01),推拿治疗后,GFAP表达下调(P<0.01),神经元形态趋于规则、排列趋于整齐,数量增加(P<0.01)。注射FCA后,星形胶质细胞抑制剂组和星形胶质细胞抑制剂+推拿组的GFAP表达水平较模型组显著降低(P<0.01),神经元形态、排列较模型组更加规整、数量增加(P<0.01)。GFAP平均荧光强度与神经元存活数量在相关性分析中提示负(r=-0.4649,P<0.05)相关。结论:推拿按揉法可以缓解CCI大鼠的疼痛行为,减轻脊髓背角神经元的受损程度,其机制可能与其抑制星形胶质细胞活化相关。Objective:To observe the effect of tuina on astrocytes and neurons in rats with chronic constriction injury(CCI),and to explore the potential mechanism of action of tuina in the treatment of neuropathic pain.Methods:Forty SD rats were randomly divided into 5 groups(n=8):control group,model group,tuina group,astrocyte inhibitor group and astrocyte inhibitor+tuina group.All groups except the control group were established CCI model.From the 4th day after modeling,the tuina group received pressing-kneading intervention for 10 minutes/time/day,and the astrocyte inhibitor group received intrathecal injection of fluorocitrate(FCA)10ul/rat/day.The astrocyte inhibitor+tuina group was treated with intrathecal injection of FCA and pressing and knead Weizhong points for 14 days.The expression of glial fibrillary acidic protein(GFAP)protein in the spinal dorsal horn was detected by immunofluorescence.The changes in the morphology,arrangement and number of neurons in the spinal dorsal horn were observed by Ni staining.The correlation between GFAP expression level and the number of surviving neurons was analyzed.Results:Compared with the control group,the mechanical pain threshold of the model group decreased significantly(P<0.01);After tuina intervention,the threshold of the tuina group gradually increased compared with the model group,and there was a significant difference after 14 days of cumulative treatment(P<0.01).From the 7th day of FCA injection,the pain threshold of the astrocyte inhibitor group and the astrocyte inhibitor+tuina group increased significantly(P<0.01).Compared with the control group,the expression of GFAP in the spinal dorsal horn of the model group was significantly increased(P<0.01),the morphology of neurons was irregular,loosely arranged,and the number of neurons was reduced(P<0.01).After tuina treatment,GFAP expression was down-regulated(P<0.01),the morphology of neurons tended to be regular,the arrangement tended to be neat,and the number of neurons increased(P<0.01).After FCA injection,GFAP expres

关 键 词：神经病理性疼痛 推拿 脊髓背角 星形胶质细胞 神经元 

分 类 号：R247.9[医药卫生—中医临床基础]