复元活血汤对大鼠损伤坐骨神经组织p-Src、p-Erk表达的影响

Effect of Fuyuan Huoxue decoction on expression of p-Src and p-Erk in injured sciatic nerve in rats

目的:研究大鼠坐骨神经离断吻合术后,复元活血汤对其修复的影响,并探讨其可能机制。方法:将60只SD大鼠实施右侧坐骨神经离断吻合术,术后大鼠随机分为复元活血汤给药组、甲钴胺给药组和模型组,每组20只。复元活血汤组给予复元活血汤,按9 g·kg^(-1)·d^(-1)灌胃;甲钴胺组给予甲钴胺,按6.25×10^(-4)g·kg^(-1)·d^(-1)灌胃;模型组按复元活血汤的溶液剂量,给予等量溶剂灌胃;灌胃2~8周。假手术组大鼠20只,暴露右侧坐骨神经后,不做任何处理缝合切口。给药3 d后,ELISA法检测大鼠血清中IL-1β和TNF-α含量,给药2周后,RT-PCR法检测吻合口近、远端2 cm神经组织的IL-1β和TNF-αmRNA的表达,Western blot法检测p-Src、p-Erk1/2蛋白的表达;分别于给药2周、4周、8周后,检测大鼠坐骨神经功能指数;于给药8周后,观察坐骨神经和其支配肌肉的病理学改变。结果:与模型组比较,复元活血汤组和甲钴胺组大鼠坐骨神经功能指数显著改善,神经纤维生长良好,腓肠肌肌纤维直径和截面积显著改善,复元活血汤组大鼠血清中IL-1β和TNF-α含量显著降低,吻合口神经组织的IL-1β和TNF-αmRNA的表达显著减少,p-Src、p-Erk1/2蛋白表达显著降低(P<0.01)。结论:复元活血汤可通过减轻大鼠损伤坐骨神经吻合口炎症反应,抑制Src、Erk的磷酸化,促进神经的修复。

AIM： To study the effect of Fuyuan Huoxue decoction on sciatic nerve regeneration in sciatic nerve injury rat model and its molecular mechanisms. METHODS： The right sciatic nerves of sixty Sprague Dawley rats were performed with transection anastomosis, and the rats were randomly divided into three groups： sciatic nerve transection anastomosis model group （Model） , Fuyuan Huoxue decoction treatment group （FHD） and mecobalamine treatment group （MB）, with twenty rats in each group postoperatively. The rats of the FHD group were fed with FHD （ig, 9 g·kg^-1·d^-1） for two to eight weeks, the rats of the MB group were fed with mecobalamine （ig, 6.25×10^-4g·kg^-1·d^-1） and the rats of the model group were fed with equivalent solvent （ig） daily. The right sciatic nerves of another twenty rats in the sham group were exposed without any treatment and then the wounds were closed. The serum IL-1β and TNF-α level of the rats were detected with ELISA method after three days of administration; the sciatic nerves functional index was investigated after two, four and eight weeks of the treatment, respectively; the mRNA expression of IL-1β and TNF-α and the proteins ex-pression of the p-Src, p-Erk1/2 of the nervous tissue 2 cm up and down anastomotic stoma were analyzed with RT-PCR and Western blot method; the pathological changes of the sciatic nerve and its innervated muscle were observed after eight weeks of the treatment. RESULTS： FHD and mecobalamine could significantly improve the sciatic nerves functional index, facilitated the nerve fibers growth, increased the diameter and cross-sectional area of the gastrocnemius muscle fiber markedly （ P 〈 0.01 ） ; FHD decreased the serum IL-1β and TNF-α level, lowed the IL-1β and TNF-α mRNA expression （P 〈 0.01 ）, the p-Src and p-Erk1/2 expression （ P 〈 0.01 ） of the nervous tissue 2 cm up and down anastomotic stoma. CONCLUSION： FHD can promote the sciatic nerve regeneration after the transection anastomosis, and the mechanism might be related to reducing inflammatory reaction, and inhibition of the Src and Erk1/2 phosphorylation in the rat sciatic nerve anastomosis.