电针干预急性脊髓损伤大鼠神经生长因子及其受体的表达变化

Effect of electro-acupuncture on expression changes of nerve growth factor and receptor in rats with acute spinal cord injury

目的:观察电针治疗后急性脊髓损伤大鼠脊髓组织神经生长因子及其受体TrkA的变化,试图发现该种变化与神经功能恢复的关系。方法:实验于2005-07-10在汕头大学医学院第二附属医院外科实验室完成。①选用2月龄SD大鼠30只,雌雄不拘。随机将大鼠分为2组:对照组和电针治疗组,每组15只。两组大鼠均采用自制的Allen装置将10g的物体从2.5cm高处落下(打击量25g·cm)致伤脊髓。对照组只造模,不进行电针治疗。电针治疗组:采用G6805-2多用治疗仪于损伤节段上、下棘突间隙,相当于第10～11胸椎和第1～2腰椎部位,各刺入一毫针,深度达硬膜外,疏密波,频率0.5ms,每天治疗30min,6d为1个疗程,间隔2d,共4个疗程。②于造模前,造模后第1天,治疗结束时采用Basso,Beattie,Bresnahan行为功能评定量表(Basso,Beattie,Bresna-hanLocomotorRatingScale,BBB)评分法(0～21分,0分:不可观察到后肢运动;21分:始终有持重的跖步,前后肢运动协调,向前时脚趾与地面之间始终保持间隙,开始触地和离地时,爪的位置与身体平行,尾巴始终上翘,躯干稳定。)评价动物运动和感觉功能。采用免疫组化方法检测两组大鼠脊髓神经元胞浆神经生长因子及其受体TrkA变化。③计量资料差异比较采用t检验。结果:大鼠30只均进入结果分析。①治疗结束时,电针治疗组神经生长因子和TrkA阳性细胞数明显多于对照组(t=3.539,2.622,P<0.01,0.05),神经生长因子和TrkA阳性细胞的平均灰度值明显低于对照组(t=6.089,6.967,P<0.01)。②造模前所有动物BBB评分均为21分,造模后第1天对照组和电针治疗组评分均为0～1分,治疗结束时,电针治疗组BBB评分明显高于对照组(t=3.847,P<0.01)。结论:电针治疗促使急性脊髓损伤大鼠脊髓神经生长因子及TrkA表达增多,促进急性脊髓损伤大鼠后肢运动和感觉功能的恢复。

AIM： To observe the changes of the nerve growth factor （NGF） and its receptor TrkA in spinal tissues of rats with acute spinal cord injury after electro-acupuncture （EA） treatment. METHODS： The experiment was finished in the Surgery Laboratory of the Second Hospital Affiliated to Medical College of Shantou University during July and October 2005.①Thirty SD rats of two-month old and either sex were divided into two groups： Control group and EA group, with 15 in each. The spinal cord injury of all the rats was induced by a 10 g × 2.5 cm impact with the self-made Allen equipment. The rats in the control group were operated only for models without EA treatment. EA group： One needle was given to the injured segment （the 10th-11^th thoracic spine） and spinous process interspace （the 1^st-2^nd lumbar vertebra） respectively by G6805-2 muhifunctional therapeutic apparatus, with the width of outside the epidural, longitudinal wave, 0.5 ms in frequency, and 30 minutes one day, six days as a course totally for four courses with the internal of two days.② Before model establishment, on the first day after model establishment and after the treatment, the motor and sensory functions of rats were tested respectively by Basso, Beattie, Bresnahan （BBB） Locomotor Rating Scale （scored from 1 to 21, 0 as non-observation of hindlimb motor, 21 as continuous coordination of sole gait, continuous toe extension, continuous extension of superior claw on the touchdown： Toes stepped on only for less than 4 times during the 4-minute observation, and were equal when lifting. The tail raised continuously but the body was unsteady.） The changes of NGF and TrkA in spinal nerves of rats in two groups were examined with im- munohistochemical method. ③The difference of measurement data were compared with t test. RESULTS： Totally 30 rats were involved in the result analysis.④After one-month EA therapy, the number of NGF and TrkA positive cells were significantly higher in EA group than in the control group （t=3.539,2.622,P 〈 0.01,0.05）, and the average gray scale of NGF and TrkA positive cells was lower in EA group than in the control group （t=6.089,6.967,P 〈 0.01）. ⑤Before the treatment, the BBB scores of all the rats were 21. Onthe first day after treatment, the scores of the control group and EA group were 0- 1. After the treatment, the score was remarkably higher in EA group than in the control group （t=3,847,P 〈 0,01）, CONCLUSION： The EA therapy can improve and increase the expression of NGF and TrkA in rats of acute spinal cord injury, and promote the rehabilitation of motor and sensory function of hindlimbs,