电针对急性脊髓损伤大鼠脊髓损伤区AMPA受体亚基GluR1的影响  

Influence of electroacupuncture on the expression of AMPA receptor subunit GluR1 in the spinal injured area of the rats with acute spinal cord injury

作　　者：陈温慈 林初勇 计静[1] 屠文展[2] 蒋松鹤[2] CHEN Wen-ci;LIN Chu-yong;JI Jing;TU Wen-zhan;JIANG Song-he(Department of Rehabilitation Medicine,Wenzhou Integrated Chinese and Western Medicine Hospital of Zhejiang Chinese Medical University,Wenzhou 325027,China;Department of Physical Medicine and Rehabilitation,Second Affiliated Hospital of Wenzhou Medical University,Wenzhou 325027,Zhejiang Province)

机构地区：[1]浙江中医药大学附属温州市中西医结合医院康复科,温州325000 [2]温州医科大学附属第二医院康复医学中心,浙江温州325027

出　　处：《中国针灸》2021年第3期307-312,共6页Chinese Acupuncture & Moxibustion

基　　金：浙江省医药卫生科研基金项目:2017KY482;温州市基础性科研项目:Y20180205。

摘　　要：目的:观察电针干预对急性脊髓损伤(SCI)大鼠AMPA受体亚基GluR1表达的影响,探讨电针在急性SCI治疗中的可能作用机制。方法:将80只SD大鼠随机分为假手术组、模型组、AMPA拮抗剂组(DNQX组)、电针组和DNQX+电针组,每组16只。采用改良Allen’s撞击法制备T10节段急性SCI大鼠模型。DNQX组于造模成功0.5 h后通过鞘内导管注射1 nmol/μL DNQX溶液10μL;电针组在造模成功后0.5、12、24 h于"大椎""命门"穴进行电针干预(疏密波,2 Hz/100 Hz,0.5 mA),每次30 min,共干预3次;DNQX+电针组同时进行上述处理。采用大鼠后肢运动功能评分(BBB)观察大鼠在造模前及造模后6、24、48 h运动功能的变化;采用HE染色观察脊髓损伤区脊髓前角组织形态变化;采用免疫荧光法检测脊髓前角GluR1阳性细胞的数量;采用Western blot法检测脊髓损伤区GluR1蛋白表达。结果:与假手术组造模后6、24、48 h相应时间点比较,模型组BBB评分均明显降低(P<0.001);各干预组BBB评分有所增加,但与模型组比较,差异无统计学意义(P>0.05)。模型组脊髓损伤区脊髓前角灰质、白质边界均模糊不清,组织间隙增大,神经元体积明显萎缩,细胞质减少、红染加深,部分神经元核固缩;电针组脊髓损伤区脊髓前角形态学改善明显,与DNQX组及DNQX+电针组相似。与假手术组比较,模型组脊髓损伤区GluR1蛋白表达增加(P<0.001),脊髓前角GluR1阳性细胞数量增多(P<0.001)。与模型组比较,电针组、DNQX组及DNQX+电针组GluR1蛋白表达减少(P<0.05,P<0.01),脊髓前角GluR1阳性细胞数量减少(P<0.001)。结论:电针干预"大椎""命门"穴可能通过抑制脊髓损伤区GluR1的表达,从而促进急性SCI大鼠脊髓前角损伤神经的修复。Objective To explore the influence of electroacupuncture(EA)on the expression of AMPA receptor subunit GluR1 in the rats with acute spinal cord injury(SCI)and explore the potential effect mechanism of EA in treatment of acute SCI.Methods A total of 80 SD rats were randomly divided into five groups,i.e.a sham-operation group,a model group,an AMPA antagonist(DNQX)group,an EA group and a DNQX+EA group,16 rats in each group.The modified Allen’s impacting method was adopted to prepare the rat model of acute SCI at T10.In the DNQX group,the intrathecal injection of 10μL DNQX solution with a concentration of 1 nmol/μL was administered in 0.5 h after modeling success.In the EA group,EA(disperse-dense wave,2 Hz/100 Hz in frequency,0.5 mA in output current)was given at"Dazhui"(GV 14)and"Mingmen"(GV 4)in 0.5 h,12 h and 24 h after modeling success for 30 min and totally 3 times.In the DNQX+EA group,the interventions in the above two groups were managed.The Basso,Beattie and Bresnahan locomotor rating score(BBB)was applied to evaluate the changes of locomotor function in the rats before modeling and in 6 h,24 h and 48 h after modeling successively.Using the hematoxylin-eosin(HE)staining,the histopathological changes in the spinal anterior horn were observed in the spinal injured area.The immunofluorescence method was adopted to determine the number of GluR1 positive neuron of the spinal anterior horn.The Western blot method was used to determine the protein expression of GluR1 in the injured area.Results Compared to the sham-operation group in 6 h,24 h and 48 h after modeling,the BBB scores were all significantly decreased in the model group(P<0.001)at the corresponding points.The BBB score was increased in each of intervention groups,but without statistical difference as compared with the model group(P>0.05).In the model group,it was found that the boundary between gray matter and white matter in the spinal anterior horn was blurred,the interstitial space enlarged,the neuron volume obviously shrunken,the cytoplasm decrea

关 键 词：急性脊髓损伤 电针 受体 AMPA GluR1亚基 谷氨酸兴奋性毒性 

分 类 号：R245.97[医药卫生—针灸推拿学]