天麻素通过调节PI3K/AKT信号通路减轻大鼠坐骨神经离断吻合后的损伤  

Gastrodin alleviate nerve injury in rats of anastomosis of transected sciatic nerve via regulating PI3K/AKT signaling pathway

作　　者：国斌 韩羽 李璐希 吴亚娟 刘雪嘉 孙广峰 聂晶 Guo Bin;Han Yu;Li Luxi;Wu Yajuan;Liu Xuejia;Sun Guangfeng;Nie Jing(Key Laboratory of Basic Pharmacology of the Ministry of Education of Zunyi Medical University and Joint Laboratory of International Cooperation of the Ministry of Education of Characteristic Ethnic Medicine,Zunyi Guizhou 563099,China;Department of Burn and Plastic Surgery,the Second Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563000,China)

机构地区：[1]遵义医科大学基础药理教育部重点实验室暨特色民族药教育部国际合作联合实验室,贵州遵义563099 [2]遵义医科大学第二附属医院烧伤整形外科,贵州遵义563000

出　　处：《遵义医科大学学报》2023年第3期234-241,共8页Journal of Zunyi Medical University

基　　金：贵州省科技计划项目,NO:黔科合基础-ZK[2022]一般629。

摘　　要：目的观察天麻素(Gas)对坐骨神经离断吻合模型大鼠神经功能修复的影响,并初步探讨其作用机制。方法32只雄性SD大鼠随机分为对照组(Control)、模型组(Model)、天麻素组(Gas)和甲钴胺组(Cob)。除Control组外,均距梨状肌下缘1 cm处锐性离断左侧坐骨神经,随即使用9-0显微外科缝合线进行端端吻合以建立坐骨神经离断吻合模型。Gas组每天灌胃给予Gas(15 mg/kg)1次,Cob组每天灌胃给予Cob(0.15 mg/kg)1次,Control组和Model组每天灌胃给予等体积双蒸水。60 d后,采用坐骨神经指数(SFI)观察坐骨神经功能恢复情况;计算腓肠肌湿重恢复率;HE与Masson染色观察坐骨神经远端有髓神经纤维的形态学变化,计数完整有髓神经纤维数;HE染色观察腓肠肌的形态学变化,统计腓肠肌横截面积;Western blot检测腓肠肌中PI3K、AKT和mTOR蛋白水平;药物亲和反应的靶点稳定性(Darts)实验检测Gas与PI3K p110 gamma的相互作用。结果与Control组相比,Model组SFI明显降低(P<0.01),腓肠肌萎缩严重,湿重显著降低(P<0.01);与Model组相比,Gas组和Cob组SFI明显升高(P<0.01,P<0.05),腓肠肌湿重增加;HE与Masson染色显示Gas和Cob可以改善坐骨神经离断所致的神经纤维排列稀疏,连续性差,增加了有髓神经纤维数量;HE染色显示Gas和Cob增加了腓肠肌纤维横截面积(P<0.01);WB结果显示,Gas和Cob增加了腓肠肌中PI3K、AKT和mTOR蛋白水平(P<0.05);药物亲和反应的靶点稳定性(Darts)实验显示,Gas可以与PI3K p110 gamma结合(P<0.01)。结论Gas通过靶向结合PI3K,上调PI3K/AKT/mTOR信号通路,发挥促进大鼠坐骨神经损伤后修复的作用。Objective To investigate the effect of gastrodin(Gastrodin,Gas)on the nerve function repair of sciatic nerve dissection anastomosis model rats and preliminarily explore its possible mechanism.Methods Thirty-two male SD rats were randomly divided into 4 groups:control group,model group,gas group and mecobalamin group.Except for the control group,transection of the left sciatic nerves conducted 1 cm under the piriformis muscle was performed in all animals,and then end-to-end anastomosis was performed with 9-0 microsurgical suture to establish the sciatic nerve dissection anastomosis model.The gas group was given gas(15 mg/kg)once a day by gavage,the mecobalamin group was given mecobalamin(0.15 mg/kg)once a day by gavage,and the control group and model group were given double distilled water of equal volume every day.After 60 days,the functional recovery was evaluated by sciatic functional index(SFI);Calculation of wet weight recovery rate of gastrocnemius muscle;Morphological changes of the distal myelinated nerve fibers of the sciatic nerve were observed by HE and Masson staining,and the number of intact myelinated nerve fibers was counted;Morphological changes of the gastrocnemius muscle were observed by HE staining,and the cross-sectional area of the gastrocnemius muscle was counted;The expression of PI3K,AKT and mTOR protein in the gastrocnemius muscle was detected by Western blot;Finally,Drug affinity responsive target stability(Darts)assay was performed to detect the interaction of Gas with PI3K p110 gamma.Results Compared with the control group,the model group had significantly lower SFI(P<0.01),severe atrophy of gastrocnemius muscle and lower wet weight(P<0.01).compared with the model group,the Gas and mecobalamin groups had significantly higher SFI(P<0.01,P<0.05)and increased wet weight of gastrocnemius muscle;HE and Masson staining showed that Gas and mecobalamin improved the sparse arrangement and poor continuity of nerve fibers due to sciatic nerve dissection and increased number of myelinated nerve fib

关 键 词：周围神经损伤 天麻素 端端吻合术 甲钴胺 

分 类 号：R745.4[医药卫生—神经病学与精神病学]