CAJ | 학술논문

目的观察脂肪来源干细胞(adipose-derive stem cells,ASCs)植入放射损伤骨骼肌后的纤维化程度改变,探讨ASCs对兔骨骼肌放射性纤维化的影响.方法 64只新西兰兔单侧臀部予9 MeV电子线单次照射80 Gy后,采用随机数字表法分为ASCs组和PBS组,每组32只,照射24 h后照射侧分别肌肉注射1 ml含5×107 ASCs的PBS悬液和1 ml PBS缓冲液,未照射一侧作为各组的正常对照.分别于照射后1、4、8和26周,收集各组8只实验动物肌肉标本,计算Masson染色切片胶原纤维面积占总面积的比值,利用免疫组织化学及Western blot法检测骨骼肌中TGF-β1蛋白的表达.结果 ASCs植入骨骼肌后能在放射损伤部位迁移.ASCs组及PBS组纤维化程度随照射后时间延长而加重,ASCs组胶原纤维面积占总面积的比值较PBS组在照射后4、8和26周显著降低(t=4.62、5.99和10.48,P＜0.05).ASCs组骨骼肌组织TGF-β1的IOD值较PBS组在照射后4、8和26周显著降低(=3.79、16.45和15.17,P＜0.05).Western blot检测ASCs组TGF-β1蛋白的表达水平较PBS组低.结论 ASCs移植能抑制放射损伤骨骼肌组织TGF-β1的表达减轻放射性纤维化.
목적 관찰지방래원간세포(adipose-derive stem cells,ASCs)식입방사손상골격기후적섬유화정도개변,탐토ASCs대토골격기방사성섬유화적영향.방법 64지신서란토단측둔부여9 MeV전자선단차조사80 Gy후,채용수궤수자표법분위ASCs조화PBS조,매조32지,조사24 h후조사측분별기육주사1 ml함5×107 ASCs적PBS현액화1 ml PBS완충액,미조사일측작위각조적정상대조.분별우조사후1、4、8화26주,수집각조8지실험동물기육표본,계산Masson염색절편효원섬유면적점총면적적비치,이용면역조직화학급Western blot법검측골격기중TGF-β1단백적표체.결과 ASCs식입골격기후능재방사손상부위천이.ASCs조급PBS조섬유화정도수조사후시간연장이가중,ASCs조효원섬유면적점총면적적비치교PBS조재조사후4、8화26주현저강저(t=4.62、5.99화10.48,P＜0.05).ASCs조골격기조직TGF-β1적IOD치교PBS조재조사후4、8화26주현저강저(=3.79、16.45화15.17,P＜0.05).Western blot검측ASCs조TGF-β1단백적표체수평교PBS조저.결론 ASCs이식능억제방사손상골격기조직TGF-β1적표체감경방사성섬유화.
Objective To investigate the effects of adipose-derived stem cell (ASC) transplantation on radiation-induced skeletal muscle fibrosis in New Zealand rabbits.Methods Totally 64 rabbits were irradiated at unilateral hip with a single dose of 80 Gy of 9 MeV electrons generated from a linear accelerator and then were randomly divided into ASCs group and PBS group with 32 abbits in each group.The non-irradiated side with normal skeletal muscle was used as control.At 24 after irradiation,1 ml PBS containing 5 × 107ASCs labbed with CM-Dil or 1 ml PBS was injected into the skeletal muscle in the irradiated side of these two groups,respectively.At 1,4,8 and 26 weeks after irradiation,the skeletal muscles of 8 rabbits in each group were collected.Masson staining was used to evaluate the level of skeletal muscle fibrosis.The expression of transforming growth factor β1 (TGF-β1) in the skeletal muscle tissues was evaluated by immunohistochemical method (IHC) and Western blot.Results The fluorescence tracing method showed that ASCs could migrate in the injured muscle and fibrosis was more serious in the PBS group.The proportion of the area of collagen fibers to the total area in sections of ASCs group was lowcr than that of PBS groups at 4,8 and 26 weeks after irradiation (t-4.62,5.99,10.48,P ＜ 0.05).The expression level of TGF-β1 protein in the ASCs group was significantly lower than that in the PBS groups at4,8 and 26 weeks after irradiation(t=3.79,16.45,15.17,P＜0.05).Conclusions ASC transplantation could alleviate radiation fibrosis by down-regulating the level of TGF-β1 protein in the irradiated skeletal muscle.