枸杞多糖干预雪旺细胞对大鼠坐骨神经损伤的实验研究  

Effect of Lycium barbarum polysaccharide on sciatic nerve injury in rats

作　　者：张博闻 马国续 王娅 赵飞[1] 李晓亮[1] 高剑 丁一 张汉霖 吴建科 黄永禄[1] Zhang Bowen;Ma Guoxu;Wang Ya;Zhao Fei;Li Xiaoliang;Gao Jian;Ding Yi;Zhang Hanlin;Wu Jianke;HuangYonglu(Department of Hand,Foot and Reconstructive Microsurgery,Ningxia Hui Autonomous Region People's Hospital,Ningxia Medical University,Yinchuan 750002,China)

机构地区：[1]宁夏回族自治区人民医院(宁夏医科大学附属自治区人民医院)手、足和重建显微外科,银川750002

出　　处：《中华手外科杂志》2024年第3期273-279,共7页Chinese Journal of Hand Surgery

基　　金：卫生部手功能重建重点实验室、上海市周围神经显微外科重点实验室开放课题(17DZ2270500);自治区科技惠民项目(2023CMG03014);自治区青年科技人才托举工程项目(2020-78);医院科技创新人才项目(2023-72)。

摘　　要：目的探讨枸杞多糖(lycium barbarum polysaccharide,LBP)干预雪旺细胞(Schwann cells,SCs)联合异种神经移植体(acellular nerve xenograft,ANX)修复大鼠坐骨神经缺损的效果.方法取大鼠的坐骨神经SCs,行原代培养,培养至第三代.将0、50、100、150、200、250、500 mg/L的LBP与SCs混合培养,分成七组.在培养皿中培养后第1、3、5、7和9天,用CCK-8法检测各组SCs生物学活性,通过rt-PCR检测各组脑源性神经营养因子(brain-derived neurotrophic factor,BDNF),神经生长因子(nerve growth factor,NGF)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)的表达水平,筛选干预SCs的LBP最适浓度.在无菌条件下制备10 mm坐骨神经缺损的大鼠模型,选取兔的胫神经制作ANX,将SD大鼠分为三组,每组10只:ANX组、ANX-SCs组和ANX-LBP-SCs组.术后8周,通过检测坐骨神经功能指数(sciatic nerve function index,SFI)、神经传导速度(motor nerve conduction velocity,MCV),应用rt-PCR检测再生神经内凋亡因子(Caspase-3)的表达水平,另通过透射电镜比较三组模型的神经移植体内的神经纤维及髓鞘再生.结果利用CCK-8测定各孔不同时间点SCs活性.在第2~4天,七组的活性差异无统计学意义(P>0.05),第6~8天150~250 mg/L LBP-SCs组增殖活性强于其他浓度组,差异有统计学意义(P<0.05).通过q-PCR检测200 mg/L LBP-SCs组中BDNF、NGF及bFGF的mRNA表达量高,与其他组相比差异有统计学意义(P<0.05).移植术后8周,发现ANX-LBP-SCs组热刺痛实验、SFI检测、神经传导速度数据更优,且Caspase-3表达量最低.透射电镜检测结果显示ANX-LBP-SCs组髓鞘水肿程度及神经纤维再生情况优于其他组.结论LBP浓度为200 mg/L时,可促进SCs的增殖,并分泌大量神经营养因子.含有LBP干预的SCs培养体系联合ANX移植可显著促进周围神经再生.Objective To investigate the effect of lycium barbarum polysaccharide(LBP)intervention on Schwann cells(SCs)combined with allogeneic nerve grafts(ANX)in repairing sciatic nerve defects in rats.Methods The sciatic nerve SCs from rats were obtained and primary culture until the third generation was performed.The 0,50,100,150,200,250,500 mg/L LBP and SCs were conducted mixed cultivation and divided into 7 groups.On the 1st,3rd,5th,7th,and 9th day after cultivation in culture dishes,the biological activity of each group of SCs was detected using the CCK-8 method.The expression levels of brain derived neurotrophic factor(BDNF),nerve growth factor(NGF),and basic fibroblast growth factor(bFGF)in each group were detected by rt-PCR,and the optimal concentration of LBP for intervention in SCs was screened.The rat model of 10 mm sciatic nerve defect was established under sterile conditions,the tibial nerve of rabbits was selected to make ANX,and SD rats were divided into three groups,with 10 rats in each group:ANX group,ANX-SCs group,and ANX-LBP-SCs group.After 8 weeks of surgery,the sciatic nerve function index(SFI)and motor nerve conduction velocity(MCV)were measured,and the expression level of Caspase-3 in the regenerated nerve was detected using rt-PCR.Additionally,the nerve fiber and myelin sheath regeneration in the nerve grafts of the three models were compared using transmission electron microscopy.Results The activity of SCs at different time points in each well was measured using CCK-8.On the 2nd to 4th day,there was no statistically significant difference in activity among the 7 groups(P>0.05),while on the 6th to 8th day,the proliferation activity of 150 to 250 mg/L LBP-SCs group was stronger than that of other concentration groups,and the difference was statistically significant(P<0.05).The mRNA expression levels of BDNF,NGF,and bFGF in the 200 mg/L LBP-SCs group were detected by q-PCR,and the differences were statistically significant compared to other groups(P<0.05).After 8 weeks of transplantation,it was foun

关 键 词：雪旺细胞 移植 枸杞多糖 异种神经移植体 

分 类 号：R285[医药卫生—中药学]