摘要
目的 探讨中药单体黄芩甙体外诱导大鼠骨髓基质细胞分化为神经细胞的可行性。方法 以黄芩甙作为主要诱导剂 ,诱导成年大鼠骨髓基质细胞 6d。采用免疫细胞化学染色、蛋白质印迹 (WesternBlot)、逆转录PCR(RT PCR)法检测神经细胞、胶质细胞标记蛋白和mRNA的表达 ;原位末端标记染色评价细胞凋亡率 ;绿色荧光蛋白转染后 ,进行同种异体脑移植 ,观察诱导后细胞在脑内存活情况。结果 黄芩甙诱导 6d后 ,骨髓基质细胞形成较典型的神经细胞形态。诱导前骨髓基质细胞不表达神经细胞、胶质细胞标记蛋白和mRNA ;诱导 6h ,nestin出现暂时表达 ,诱导 6d表达神经细胞标记蛋白和mRNA ,不表达胶质细胞标记蛋白和mRNA。细胞凋亡率为 12 2 %± 2 8%。同种异体脑移植 14d ,移植物存活良好。结论 黄芩甙可以定向诱导骨髓基质细胞分化为神经细胞。
Objective To study the possibilty that induced marrow stromal cells (MSCs) to neuron in vitro by baicalin, a kind of flavonoid isolated from an important medicinal plant Scutellariae Radix Methods MSCs from adult SD rats were induced by baicalin in serum-free medium for 6 days The expression of neuronal or glia specific markers and gene was evaluated by indirect immunofluorescence cytochemistry staining,Western blot,and reverse transcriptase -polymerase chain reaction (RT-PCR) The percentage of apoptotic cells at 6 days after baicalin treatment was measured by terminal deoxynucleotidyl transferase- mediated dUTP-biotin nick end-labeling (TUNEL) assay Moreover, differentiating cells that labeled by green fluorescent protein vector were transplanted to the striatum of adult SD rats after 6d of baicalin treatment Results After induction for 6d, MSCs displayed neuronal morphologies, such as pyramidal cell bodies and processes formed extensive networks The undifferentiated cells did not exhibit a neuronal or glial phenotype The differentiating cells expressed nestin, a marker of neural stem cells, at 6 h, and disappeared at 6 d In contrast, expression of neuron-specific markers and gene was detectable at 6 d And glia-specific markers and gene was not expressed The percentage of apoptotic cells was 12 2%±2 8% In addition, fluorescently labeled cells were readily detected in the brain section after 14d of transplantation Conclusion Baicalin may induce adult rat MSCs to neuron in vitro It is a useful vehicles for autotransplantation in both cell and gene therapy for a variety of diseases of the central nervous system
出处
《中华医学杂志》
CAS
CSCD
北大核心
2002年第19期1337-1341,共5页
National Medical Journal of China
基金
国家自然科学基金资助项目 (No .3 9870 2 5 1)
中国博士后科学基金资助项目 (第 2 9期 )
