督脉电针治疗脊髓损伤后不同神经营养因子表达的时间窗特征

Characteristics of time window of the expression of different neurotrophic factors after the treatment of electro-acupuncture on Du Channel after spinal cord injury

目的:观察脊髓损伤后督脉电针治疗对内源性神经营养因子家族中神经生长因子、脑源性神经营养因子和成纤维细胞生长因子-2表达的影响。方法:实验于2003-06/2004-06在滨州医学院人体解剖学教研室完成。选用成年雌性Wistar大鼠72只,采用Allen法制备脊髓打击损伤模型,72只大鼠随机分为3组,每组24只。每组再分为3d、1,2,4周4个时间点,每个时间点6只。①电针组:损伤后即刻开始电针治疗,取督脉“大椎”和“命门”穴,1.65cm(0.5寸)毫针,深达硬膜外。然后用多功能治疗仪治疗,强度以大鼠后肢微微颤动为度。1次/d,30min/次。②激素组:损伤后即刻前肢皮下注射甲基强的松龙(80mg/kg);此后分别于6,12,18h再次注射(40mg/kg)。③损伤组:损伤后不做治疗。术后3d、1周、2周和4周应用免疫组织化学染色分别观察损伤脊髓神经生长因子、脑源性神经营养因子和成纤维细胞生长因子-2变化。结果:72只大鼠全部进入结果分析。①神经生长因子表达变化最显著,损伤组和电针组3d时阳性细胞增多,至2周时最多,4周时明显减少,电针组明显多于损伤组。激素组3d达高峰,此后呈下降趋势,4周时回落到损伤组水平,阳性反应较电针组稍弱,比损伤组明显增强。②脑源性神经营养因子表达:电针组3d时就明显高于损伤组,而且持续整个实验过程;激素组在1周时脑源性神经营养因子反应明显增强,2周后降至损伤组水平。③成纤维细胞生长因子-2表达:脊髓损伤后成纤维细胞生长因子-2表达局限于前2周,比神经生长因子、脑源性神经营养因子表达弱,持续时间短,电针组和激素组明显多于损伤组,2周后与损伤组已无明显差异。结论:督脉电针治疗能促进损伤脊髓内源性神经营养因子的表达,不同的因子变化时间窗不同。

AIM： To observe the effects of Du Channel electro-acupuncture on the expression of nerve growth factor （NGF）, brain-derived neurotrophic factor （BDNF） and fibrous growth factor-2 （FGF-2） in the endogenous neurotrophic factor family after spinal cord injury. METHODS： The experiment was done in the Department of Anatomy of Binzhou Medical College from June 2003 to June 2004. Seventy-two female adult Wistar rats were used to make spinal cord injury model by Allen＇s method, and were assigned randomly into 3 groups with 24 rats in each group. Then the rats in each group were divided into 4 subgroups of 3 days, 1, 2, 4, weeks with 6 rats in each subgroup. ① Electro-acupuncture group： The electro-acupuncture was performed after injury immediately. Dazhui （DU 14） and Mingmen （Du 4） of Du Channel were treated with 1.65 cm （0.5 inch） filiform needle reaching deeply to epidural, and then treated with multi-functional treatment apparatus in the intensity of the slight tremor of rat＇s hindlimb with once a day and 30 minutes every time. ② Hormones group： Artisone-wyeth was injected subcutaneously into forelimb after injury instantly （80 mg/kg）; After that at 6, 12, and 18 hours injected （40 mg/kg） again respectively. ③ Injured group： Without treatment after injury, the changes of injured spinal nerves growth factor, BDNF and FGF-2 were observed respectively by immunohistochemistry staining 3 days, 1 week, 2 weeks, and 4 weeks after operation. RESULTS： Totally 72 rats were involved in the result analysis. ① The change of NGF expression was the most significance. The positive cells increased at 3 days in injured and electro-acupuncture group, and the maximum at 2 weeks and significantly decreased at 4 weeks, and it was significantly more in electric needle group than that in injured group. It reached the peak at 3 days in hormones group, showed the trend of decrease later, and went back to the level in injured group at 4 weeks. The positive reaction was a little weaker than that in electro-acupuncture and significantly stronger than that in injured group. ② The expression of BDNF： It was significantly higher in electro-acupuncture group than that in injured group at 3 days, and lasted during the whole process of experiment; BDNF reaction significantly increased at 1 week in hormones group, and decreased to the level of injured group after 2 weeks. ③ The expression of FGF-2： It was limited to the former two weeks after the spinal injury, and weaker than that of NGF and BDNF and the lasting time was short. It was more in electric needle and hormones group than that of injured group, and there was no significant difference compared with injured group after 2 weeks. CONCLUSION： Treatment with electro-acupuncture on Du Channel can improve the expression of endogenous neurotrophic factors after spinal cord injury, and the time window of different factors is different.