夹脊电针对急性脊髓损伤大鼠脊髓组织微环境Rho-ROCK Ⅱ通路相关因子的影响  

Electroacupuncture Improves Limb Locomotor Function Possibly by Suppressing Rho-ROCK Ⅱ Pathway Related Factors in Anterior Horns of Spinal Cord in Rats with Acute Spinal Cord Injury

作　　者：李晓宁[1] 梁雪松[2] 吴磊[2] 单筱淳 付豪[2] 梅继林 李诺[2] LI Xiao-ning;LIANG Xue-song;WU Lei;SHAN Xiao-chun;FU Hao;MEI Ji-lin;LI Nuo(The Fourth Department of Acupuncture and Moxibustion,the Second Affiliated Hospital of Heilongjiang University of Traditional Chinese Medi-cine,Harbin 150001,China;The Second Clinical College of Heilongjiang University of Traditional Chinese Medicine,Harbin 150040)

机构地区：[1]黑龙江中医药大学附属第二医院针灸四科,哈尔滨150001 [2]黑龙江中医药大学第二临床医学院,哈尔滨150040

出　　处：《针刺研究》2018年第7期445-449,455,共6页Acupuncture Research

基　　金：国家自然科学基金项目(No.81373715);哈尔滨市应用技术研究与开发项目科技创新人才类(No.2016RAXYJ 090)

摘　　要：目的:观察夹脊电针对脊髓损伤大鼠神经细胞轴突再生相关抑制因子重组人Ras同源物基因家族成员A(RhoA)、Rho蛋白激酶Ⅱ(ROCKⅡ)、肌球蛋白轻链(MLC)蛋白的影响,探讨夹脊电针治疗急性脊髓损伤(ASCI)的作用机制。方法:雌性Wistar大鼠随机分为假手术组、模型组、夹脊电针组、抑制剂组,每组12只。采用脊髓打击法制备大鼠ASCI模型。夹脊电针组于模型制备成功后3h进行夹脊电针治疗,每日1次,每次30 min,连续治疗14d、28d;抑制剂组于造模成功后立即给予腹腔注射盐酸法舒地尔(10mg/kg),每日1次,连续治疗14d、28d。采用BBB评分法评估大鼠后肢运动功能变化,免疫组化法检测各组大鼠脊髓组织中RhoA、ROCKⅡ、MLC蛋白的表达情况。结果:BBB评分结果显示:与假手术组比较,造模后14d、28d模型组大鼠运动功能评分显著降低(P<0.05);与模型组比较,造模后14d、28d夹脊电针组、抑制剂组大鼠运动功能评分均显著升高(P<0.05);夹脊电针组、抑制剂组造模后28d大鼠肢体运动功能评分较造模后14d显著升高(P<0.05)。免疫组化结果显示:与假手术组比较,造模后14d、28d模型组大鼠脊髓组织RhoA、ROCKⅡ、MLC阳性细胞数增加(P<0.05);与模型组相比,造模后14d、28d夹脊电针组、抑制剂组大鼠脊髓组织RhoA、ROCKⅡ、MLC阳性细胞数显著降低(P<0.05);夹脊电针组、抑制剂组造模后28d大鼠脊髓组织RhoA、ROCKⅡ、MLC阳性细胞数较造模后14d显著降低(P<0.05)。结论:夹脊电针能够显著改善ASCI大鼠肢体运动功能,其作用机制可能与抑制大鼠脊髓损伤组织微环境Rho-ROCKⅡ信号通路RhoA、ROCKⅡ、MLC抑制因子表达有关。Objective To observe the effect of electroacupuncture（EA）of＂Jiaji＂（EX-B 2）on limb locomotor function and expression of Ras homolog gene family member A（RhoA）,Rho-associated kinase Ⅱ（ROCK Ⅱ）and myosin light chain（MLC）proteins in the anterior horn of spinal cord in acute spinal cord injury（ASCI）rats,so as to explore its mechanisms underlying improvement of SCI-induced limb locomotor dysfunction.Methods Forty-eight female Wistar rats were randomly divided into sham operation（sham）,ASCI model（model）,EA EX-B 2（EA）and ROCK inhibitor（Fasudil）groups which were further divided into 14 dand 28 dsubgroups（n=6 in each）.The ASCI model was made by using weight drop striking method.Three hours after modeling,EA（100 Hz,0.4,0.6 mA）was applied to EX-B 2（T 9,T 11）for 30 min,once daily for 14 dand 28 d,respectively.The ROCK inhibitor（hydrochloride Fasudil,10 mg/kg）was administrated by intraperitoneal injection immediately after modeling,once a day,continuously for 14 dor 28 d.The expression of RhoA,ROCK Ⅱ and MLC proteins in the spinal cord anterior horn tissue（T 10）was detected by immunohistochemistry.The rats＇ hindlimb locomotor function was assessed according to Basso,Beattie and Bresnahan（BBB）locomotor rating scale（21-points）.Results After ASCI,the BBB scores were significantly lower in the model group than in the sham group on day 14 and 28（P〈0.05）,and obviously higher in the EA and inhibitor groups than in the model group（P〈0.05）,suggesting an improvement of the hindlimb locomotor function after EA intervention or suppression of ROCK.Immunohistochemical results indicated that the numbers of RhoA,ROCK Ⅱ and MLC immune-reaction positive cells in the anterior horn of spinal cord were significantly more in the model group than in the sham group（P〈0.05）,and remarkably decreased in both EA and inhibitor groups on day 14 and 28 relevant to the model group（P〈0.05）.The therapeutic effects of EA were markedly weaker than those of in

关 键 词：急性脊髓损伤 电针 髓鞘相关抑制因子 Rho-ROCK Ⅱ信号通路 重组人Ras同源物基因家族成员A Rho蛋白激酶Ⅱ 肌球蛋白轻链 

分 类 号：R245.97[医药卫生—针灸推拿学]