中华骨科杂志
中華骨科雜誌
중화골과잡지
CHINESE JOURNAL OF ORTHOPAEDICS
2013年
5期
561-568
,共8页
刘燊%冯世庆%周先虎%宁广智%Ying Sun%Xin-fu Zhou
劉燊%馮世慶%週先虎%寧廣智%Ying Sun%Xin-fu Zhou
류신%풍세경%주선호%저엄지%Ying Sun%Xin-fu Zhou
脑源性神经营养因子%脊髓损伤%少突神经胶质
腦源性神經營養因子%脊髓損傷%少突神經膠質
뇌원성신경영양인자%척수손상%소돌신경효질
Brain-derived neurotrophic factor%Spinal cord injuries%Oligodendroglia
目的 观察脑源性神经营养因子前体(progenitor of Brain-derive neurotrophic factor,proBDNF)对少突胶质细胞前体细胞(oligodendrocyte precusor cells,OPCs)增殖、分裂及迁移的影响,探讨proBDNF与p75神经营养因子受体(p75 neurotrophic receptor,p75NTR)信号转导通路的关系.方法 体外实验应用大鼠少突胶质细胞系OLN-93作为研究对象,测定不同浓度proBDNF对OLN-93细胞分裂、增殖活性的影响；应用免疫荧光染色方法观察OLN-93细胞表面p75NTR 、sortilin和内源性proBDNF 的表达；观察proBDNF抗体治疗对OPCs增殖活性的影响.体内实验应用proBDNF抗体对SD大鼠T9脊髓损伤模型进行治疗,观察BBB评分的改善情况；应用免疫荧光染色观察BrdU+/NG2+细胞在脊髓损伤后的数量及聚集部位,评估proBDNF及其抗体对OPCs生物学活性的影响,并观察p75NTR及sortilin 在治疗前后的表达水平变化.结果 proBDNF对OPCs的分裂、增殖具有明显的抑制作用,且可被proBDNF抗体所拮抗.p75NTR的表达水平可被proBDNF抗体下调.结论 内源性proBDNF对OPCs的分裂、增殖具有明显的抑制作用,极有可能通过p75NTR-sortilin通路介导.proBDNF抗体治疗可以拮抗proBDNF的作用,提高OPCs的活性并促进损伤后的功能恢复,具有重要的研究与应用价值.
目的 觀察腦源性神經營養因子前體(progenitor of Brain-derive neurotrophic factor,proBDNF)對少突膠質細胞前體細胞(oligodendrocyte precusor cells,OPCs)增殖、分裂及遷移的影響,探討proBDNF與p75神經營養因子受體(p75 neurotrophic receptor,p75NTR)信號轉導通路的關繫.方法 體外實驗應用大鼠少突膠質細胞繫OLN-93作為研究對象,測定不同濃度proBDNF對OLN-93細胞分裂、增殖活性的影響；應用免疫熒光染色方法觀察OLN-93細胞錶麵p75NTR 、sortilin和內源性proBDNF 的錶達；觀察proBDNF抗體治療對OPCs增殖活性的影響.體內實驗應用proBDNF抗體對SD大鼠T9脊髓損傷模型進行治療,觀察BBB評分的改善情況；應用免疫熒光染色觀察BrdU+/NG2+細胞在脊髓損傷後的數量及聚集部位,評估proBDNF及其抗體對OPCs生物學活性的影響,併觀察p75NTR及sortilin 在治療前後的錶達水平變化.結果 proBDNF對OPCs的分裂、增殖具有明顯的抑製作用,且可被proBDNF抗體所拮抗.p75NTR的錶達水平可被proBDNF抗體下調.結論 內源性proBDNF對OPCs的分裂、增殖具有明顯的抑製作用,極有可能通過p75NTR-sortilin通路介導.proBDNF抗體治療可以拮抗proBDNF的作用,提高OPCs的活性併促進損傷後的功能恢複,具有重要的研究與應用價值.
목적 관찰뇌원성신경영양인자전체(progenitor of Brain-derive neurotrophic factor,proBDNF)대소돌효질세포전체세포(oligodendrocyte precusor cells,OPCs)증식、분렬급천이적영향,탐토proBDNF여p75신경영양인자수체(p75 neurotrophic receptor,p75NTR)신호전도통로적관계.방법 체외실험응용대서소돌효질세포계OLN-93작위연구대상,측정불동농도proBDNF대OLN-93세포분렬、증식활성적영향；응용면역형광염색방법관찰OLN-93세포표면p75NTR 、sortilin화내원성proBDNF 적표체；관찰proBDNF항체치료대OPCs증식활성적영향.체내실험응용proBDNF항체대SD대서T9척수손상모형진행치료,관찰BBB평분적개선정황；응용면역형광염색관찰BrdU+/NG2+세포재척수손상후적수량급취집부위,평고proBDNF급기항체대OPCs생물학활성적영향,병관찰p75NTR급sortilin 재치료전후적표체수평변화.결과 proBDNF대OPCs적분렬、증식구유명현적억제작용,차가피proBDNF항체소길항.p75NTR적표체수평가피proBDNF항체하조.결론 내원성proBDNF대OPCs적분렬、증식구유명현적억제작용,겁유가능통과p75NTR-sortilin통로개도.proBDNF항체치료가이길항proBDNF적작용,제고OPCs적활성병촉진손상후적공능회복,구유중요적연구여응용개치.
Objective To observe whether immature Brain Derived Neurotrophic Factor (proBDNF)can affect the activities of OPCs in the fields of cell proliferation and migration after SCI,and to investigate the relationship between proBDNF and p75NTR signal pathway on OPCs.Methods OLN-93 cell line was cultured and maintained for in vitro experiments.Immunofluorescence were used to check the expression of endogenous proBDNF,p75NTR and sortilin on OPCs.MTT assay was used to illustrate the inhibitory effect of proBDNF.The effects of anti-proBDNF was also observed by BrdU staining to find a probably signal pathway for proBDNF on OPCs.The Sprague-Dawley rats were administered for T9 spinal cord injury animal model.BBB score was applied to observe the situation of functional recovery after treated by anti-proBDNF.BrdU staining was managed to observe the situation of OPCs proliferation and migration after SCI.Results Endogenous proBDNF inhibited proliferation and migration of OPCs after SCI.BrdU staining showed that population of proliferative OPCs in lesion site of spinal cord was less in proBDNF in treated group than that in control group and anti-proBDNF group.While anti-proBDNF could inhibit proBDNF specifically and might induced a better functional recovery which was illustrated by BBB scores.The in vitro experiments found the inhibitory effect of proBDNF is dose-dependent and can be neutralized by anti-proBDNF properly.Moreover,the expression levels of p75NTR and sortilin are down regulated by proBDNF antibody treated group.This indicated that proBDNF may inhibit OPCs via p75NTR pathway.Conclusion Endogenous proBDNF can inhibit cell proliferation of OPCs after SCI and can be neutralized by specific antibodies of proBDNF.This kind of detrimental effect may be induced by p75NTR-sortilin pathway.Furthermore,proBDNF antibody treatment is effective to block proBDNF and promote the functional recovery.
