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作 者:冷雁[1] 张为西[1] 周琛[1] 郑振扬[1] 张成[1] 李秋玲[1]
机构地区:[1]中山大学附属第一医院神经科,广东省广州市510080
出 处:《中国组织工程研究与临床康复》2011年第33期6173-6176,共4页Journal of Clinical Rehabilitative Tissue Engineering Research
基 金:国家自然科学基金(30870852;30971026)~~
摘 要:背景:干细胞移植是治疗肌营养不良症的有效方法之一,但移植的干细胞在病理骨骼肌中成肌表达较低。目的:通过比较mdx小鼠和C57小鼠的骨骼肌形态及成肌、成脂、成骨基因表达的差异,探讨mdx小鼠骨骼肌病理改变的可能机制。方法:取mdx小鼠与C57小鼠的骨骼肌组织行冰冻切片,苏木精-伊红染色和Vonkossa染色观察两种小鼠肌肉组织的形态特征;提取mdx小鼠和C57小鼠骨骼肌组织总RNA,real-timePCR检测成肌、成脂、成骨相关基因的表达。结果与结论:mdx小鼠骨骼肌有肌纤维坏死和再生,伴有轻度脂肪、纤维结缔组织增生,Vonkossa染色可见钙结节沉积,而C57小鼠的骨骼肌细胞形态清晰,核位于细胞周边。与C57小鼠比较,mdx小鼠肌肉组织成骨、成脂基因表达有不同程度的上调(P<0.05),而成肌基因表达下调(P<0.05)。dystrophin基因缺失及成肌基因表达下调、成骨和成脂基因上调是造成mdx小鼠肌肉组织变性坏死的原因。BACKGROUND:Duchenne's muscular dystrophy(DMD)is a fatal recessive X-linked form of muscular dystrophy characterized by progressive muscular degeneration,and there is no cure for DMD currently.Stem cell transplantation may provide us with a creative perspective.But the myogenic differentiation rate of transplanted cells is quite low in skeletal muscle.OBJECTIVE:The differences of myogenic,adipogenic and osteogenic gene expression levels in skeletal muscle between mdx mice and C57BL/6J mice were compared,with the purpose of investigating the underlying mechanism of pathological changes in skeletal muscle from mdx mice.METHODS:Frozen sections of skeletal muscle from mdx and C57BL/6J mice were prepared,stained with hematoxylin-eosin staining and Vonkossa staining.The morphological changes of the muscles were observed under microscope.Additionally,total RNA of skeletal muscle from mdx and C57BL/6J mice was extracted,reversed,and the expression levels of myogenic,adipogenic and osteogenic characteristic genes were examined by real-time PCR.RESULTS AND CONCLUSION:Necrosis and regeneration were found in skeletal muscles of mdx mice,and mild adipose hyperplasia and fibrous connective tissue hyperplasia were also observed.Moreover,calcium deposition nodules were easily detected by Vonkossa staining.The form of skeletal muscle cells from C57 mice was clear,and the nuclei were located in the cell periphery.Osteogenic and adipogenic gene expressions of skeletal muscle from mdx mice were elevated to a certain degree by real-time PCR(P 0.05),compared with C57 mice,whereas myogenic gene expression was decreased(P 0.05).The reason why adipocyte and osteoblast in skeletal muscle of mdx mice overgrew may be due to degeneration and necrosis of skeletal muscle which caused by dystrophin gene deletion,and it differentiates into osteoblasts and adipocytes instead of myoblasts.
关 键 词:肌营养不良症 mdx小鼠 DYSTROPHIN基因 骨骼肌 基因表达
分 类 号:R318[医药卫生—生物医学工程]
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