神经生长因子及三磷酸腺苷联合应用修复大鼠周围神经损伤及对失神经肌肉的作用

目的 探讨腹腔联合应用鼠源性神经生长因子(NGF)及三磷酸腺苷(ATP)对周围神经损伤修复及失神经肌肉的作用. 方法 选取SD大白鼠96只,体重(250±15)g,雌雄不限,按随机数字表法分为等渗盐水组(NS组)、ATP组、NGF组、ATP+NGF组;选取大鼠坐骨神经,暴露后人为切断造成3 mm缺损损伤,通过硅胶管连接两神经断端,建立神经再生室.术后立即对各组分别予以腹腔内注射等渗盐水0.4 ml、ATP0.4 ml(1 mg/ml)、NGF0.4 ml(0.2 μg/ml)、NGF+ATP 0.4 ml(ATP 1 mg/ml、NGF 0.2 μg/ml),之后每3 d通过腹腔注射1次.在第2,4,6,8周各组分别取6只大门鼠行坐骨神经指数测定、大体观察、坐骨神经肌电图检查,处死大鼠后切取标本观察再生室区段神经再生状况、腓肠肌湿重,HE染色观察腓肠肌纤维直径及截面积. 结果 术后第2,4,6,8周各组分别取6只大白鼠测定实验数据,NGF+ATP组坐骨神经指数、神经传导速度、腓肠肌湿重优于NS组及单纯ATP或NGF组(P＜0.05).大体观察可见NGF+ATP组患侧足趾红肿程度较其余三组轻,局部溃烂亦相对少,未见足趾脱落者.术后2周,各组神经再生室内可见纤细神经纤维生长,但未将神经断端连接一起.术后4,6,8周,神经再生室内可见神经纤维通过,NGF+ATP组相对其余三组神经纤维生长生长情况较好,且局部炎性反应及纤维粘连轻.NGF组神经纤维生长情况好于ATP组,但二者均优于NS组.第2,4,6,8周神经电生理及腓肠肌湿重NGF组优于ATP组,第4,6,8周坐骨神经指数、肌纤维直径及截面积NGF组优于ATP组(P＜0.05),且各时相点NGF组及ATP组的观测指标均明显优于NS组(P＜0.05). 结论 (1)联合应用NGF+ATP对神经损伤修复及失神经肌肉作用明显,优于单纯应用NGF及ATP;(2)单纯应用ATP对神经损伤修复及失神经肌肉有一定作用,但效果劣于NGF及NGF+ATP.

Combined used of nerve growth factor and adenosine triphosphate for repair of peripheral nerve injury and its influence on the denervated muscle

Objective To investigate the unification effect of rat nerve growth factor(NGF)and adenosine triphosphate(ATP)on repair of the peripheral nerve injury and the denervated muscle.Methods Ninety-six healthy SD rats weighing(250±15)g were randomly divided into NGF+ATP group,NGF group,ATP group and NS group.The sciatic nerves were cut for 3 mm,and were connected with silicone tube to create the nerve regeneration room.Then,all rats were intraperitoneally injected with NS0.4ml,ATP0.4ml(1 mg/ml),NGF 0.4 ml(0.2 μg/ml),NGF+ATP 0.4 ml(ATP1 mg/ml,NGF 0.2 μg/ml)separately once every three days.Six rats from each group were selected at 2,4,6 and 8 weeks after operation for sciatic nerve index(SFI)detection,gross observation and electrophysiological test;the specimens were obtained from the sacrificed rats for nerve regeneration,gastrocnemius muscle wet weight,fiber diameter and cross-sectional area of the gastrocnemius muscle(by HE staining).ResultsAt 2,4,6 and 8 weeks after operation,every six rats were chosen to be tested.The SFI,nerve conduction velocity and wet weight of the muscle of NGF+ATP group were better than those of the other three groups(P ＜ 0.05).The swelling of the rat toes of NGF+ATP group was slighter than that of NGF group and ATP group,with no loss of toes found in NGF+ATP group.At the 2nd week after operation,new nerve fibers were observed in the regeneration room,with no connection of the nerve fibers.The nerve fibers crossed the regeneration room at 4,6 and 8 weeks after operation.Compared with other three groups,the nerve growth condition was better in NGF+ATP group,with slighter adhesion and inflammation of nerve fibers.The growth condition of the nerve fibers of NGF group was better than ATP group,and that of NGF group and ATP group was better than NS group.At 2,4,6 and 8 weeks after operation,the electrophysiological condition and wet weight of muscle in NGF group was better than those in ATP group.While the SFI,the cross area and diameter of the muscle in NGF group were better than ATP group at 4,6 sand 8 weeks(P ＜ 0.05).All the parameters in NGF group and ATP group were better than NS group at each time point(P ＜ 0.05).Conclusions(1)The unification effect of NGF and ATP exert apparent effect in repair of the injured nerve and on the denervated muscle.Combined application of NGF and ATP is superior than NGF or ATP used alone.(2)A certain effect of ATP in repair of the injured nerve and on the denervated muscle is detected,but it is inferior to NGF and NGF+ATP.