雷公藤甲素干预脊髓损伤模型Thy-YFP转基因小鼠调节自噬及抑制细胞凋亡  

Triptolide improves spinal cord injury recovery via upregulation of autophagy and inhibition of apoptosis in Thy-YFP transgenic mice

作　　者：朱宁 杨新明[1] 阮建伟 Zhu Ning;Yang Xinming;Ruan Jianwei(First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,Hebei Province,China)

机构地区：[1]河北北方学院附属第一医院,河北省张家口市075000

出　　处：《中国组织工程研究》2020年第35期5650-5655,共6页Chinese Journal of Tissue Engineering Research

基　　金：河北省卫生厅2011 年医学科学研究重点项目计划(20110176),项目负责人:杨新明;2013 年度河北北方学院创新人才培育基金项目(CXRC1322),项目负责人:杨新明。

摘　　要：背景:研究表明,在脑卒中大鼠模型中,雷公藤甲素治疗减少了缺血性病变区域面积、水含量和神经细胞死亡。此外,雷公藤甲素能通过抑制星形胶质细胞增生、小胶质细胞活化和抑制炎症反应促进脊髓损伤修复。目的:研究雷公藤甲素对Thy-YFP转基因小鼠脊髓损伤后细胞自噬和细胞凋亡的影响,探讨雷公藤甲素对脊髓损伤的保护作用及机制。方法:将60只Thy-YFP转基因小鼠随机分为4组,分别是假手术组、二甲基亚砜组、甲基强的松龙组和雷公藤甲素组。假手术组只进行椎板切除术,不损伤脊髓;其他3组建立脊髓损伤模型。雷公藤甲素组、二甲基亚砜及假手术组小鼠术后立即分别腹腔注射雷公藤甲素[0.2 mg/(kg·d)]或等量5%二甲基亚砜-生理盐水溶液,连续给药7 d;甲基强的松龙组小鼠于术后30 min、6 h、24 h腹腔注射甲基强的松龙溶液(30 mg/kg)。BMS评分检测小鼠运动功能恢复情况,苏木精-染色及Nissl染色法检测各组脊髓组织恢复情况,免疫印迹及免疫荧光染色检测各脊髓组织中自噬相关蛋白Beclin-1、LC3B、p62及凋亡相关蛋白Bcl-2、Bax、caspase-3的蛋白水平。动物伦理获得河北北方学院批准(W20200002)。结果与结论:①雷公藤甲素及甲基强的松龙组脊髓损伤后运动功能改善,神经细胞死亡减少;②雷公藤甲素处理后自噬相关蛋白LC3B上调,p62降低,细胞凋亡相关蛋白caspase-3和Bax降低,抗凋亡蛋白Bcl-2升高;③结果表明,雷公藤甲素能促进脊髓损伤后运动功能恢复,其机制与促进自噬和抑制凋亡有关。结果提示雷公藤甲素可能对脊髓损伤具有潜在的神经保护作用。BACKGROUND:Studies have shown that treatment with triptolide reduces ischemic lesion area,water content,and nerve cell death in a rat model of stroke.In addition,triptolide can promote the repair of spinal cord injury by inhibiting astrocyte hyperplasia,microglia activation and inflammatory response.OBJECTIVE:To investigate the effect of triptolide on the autophagy and apoptosis in the spinal cord of Thy-YFP transgenic mice after spinal cord injury.METHODS:Sixty Thy-YFP transgenic mice were randomly divided into four groups:sham,dimethyl sulfoxide(DMSO),triptolide and methylprednisolone groups.Following the establishment of the spinal cord injury models,mice in the triptolide,DMSO and sham group were immediately intraperitoneally injected with triptolide(0.2 mg/kg·d)or 5%DMSO-normal saline solution(0.2 mg/kg·d)for 7 days,whereas mice in the methylprednisolone group were intraperitoneally injected with methylprednisolone(30 mg/kg)at 30 minutes,6 hours and 24 hours after surgery.Basso Mouse Scale was used to assess the recovery of motor function;hematoxylin-eosin staining and Nissl staining were used to explore tissue recovery after spinal cord injury;western blot and immunofluorescence staining were employed to assess the levels of autophagy-associated proteins,Beclin-1,LC3B,p62,and apoptosis-associated proteins,Bcl-2,Bax and caspase-3.An ethic approval for animal experiments was obtained from Hebei North University with an approval No.W20200002.RESULTS AND CONCLUSION:Treatments with triptolide or methylprednisolone after spinal cord injury significantly improved motor function and reduced neuronal cell death.In addition,the expression of autophagy-associated protein LC3B was upregulated and the expression of p62 was downregulated following triptolide treatment;the expressions of apoptosis-associated proteins caspase-3 and Bax were reduced,while the expression of anti-apoptosis protein Bcl-2 was increased after triptolide treatment.To conclude,triptolide improves the motor function after spinal cord injury,which

关 键 词：雷公藤甲素 自噬 凋亡 脊髓损伤 动物 小鼠 实验 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]