CAJ | 학술논문

目的：观察黄芪多糖对大鼠骨髓间充质干细胞(BMSCs)向神经细胞、脂肪细胞、成骨细胞和软骨细胞分化特性的影响,为开发有效而又低毒的分化诱导剂提供依据。方法采用全骨髓贴壁筛选法分离、纯化无特定病原体级Wistar大鼠BMSCs。用噻唑蓝(MTT)法筛选出黄芪多糖的合适浓度,取F3代细胞,采用随机数字表法分为对照组和诱导组(神经诱导、成脂诱导、成骨诱导、软骨诱导),采用甲苯胺蓝染色、油红O染色、茜素红染色检测神经细胞、脂肪细胞、软骨细胞表面特异性标记物。用Western Blot技术检测神经元特异性烯醇化酶(NSE)、脂蛋白酯酶(LPL)、胶原蛋白Ⅰ、胶原蛋白Ⅱ。分析黄芪多糖对F3代BMSCs向神经、脂肪、软骨和骨的诱导分化作用。结果 MTT显示,在1 g/L黄芪多糖诱导48 h下细胞增殖明显,甲苯胺蓝染色阳性,而油红O、茜素红染色阴性。Western Blot法检测显示NSE为阳性表达,LPL、胶原蛋白Ⅰ、胶原蛋白Ⅱ均为阴性表达。结论黄芪多糖可诱导大鼠BMSCs定向分化为神经细胞,而未向脂肪细胞、成骨细胞和软骨细胞分化。
목적：관찰황기다당대대서골수간충질간세포(BMSCs)향신경세포、지방세포、성골세포화연골세포분화특성적영향,위개발유효이우저독적분화유도제제공의거。방법채용전골수첩벽사선법분리、순화무특정병원체급Wistar대서BMSCs。용새서람(MTT)법사선출황기다당적합괄농도,취F3대세포,채용수궤수자표법분위대조조화유도조(신경유도、성지유도、성골유도、연골유도),채용갑분알람염색、유홍O염색、천소홍염색검측신경세포、지방세포、연골세포표면특이성표기물。용Western Blot기술검측신경원특이성희순화매(NSE)、지단백지매(LPL)、효원단백Ⅰ、효원단백Ⅱ。분석황기다당대F3대BMSCs향신경、지방、연골화골적유도분화작용。결과 MTT현시,재1 g/L황기다당유도48 h하세포증식명현,갑분알람염색양성,이유홍O、천소홍염색음성。Western Blot법검측현시NSE위양성표체,LPL、효원단백Ⅰ、효원단백Ⅱ균위음성표체。결론황기다당가유도대서BMSCs정향분화위신경세포,이미향지방세포、성골세포화연골세포분화。
Objective To investigate the effect of astragalus polysaccharides (APS) on the differentiation of rat bone marrow mesenchymal stem cells (BMSCs) to neurones, adipocytes, osteoblasts and chondrocytes, and provide basis for the development of effective and low toxic differentiation inducing agents. Methods BMSCs were isolated from SPF Wistar rats, purified, expanded and cultured to family 3. The appropriate concentration of APS was filtered out by MTT assay. The F3 cells were randomly divided into control group and induced group (neural induction, adipogenic induction, osteogenic induction, cartilage induction). The effects of APS and classical chemical drugs on differentiation were measured by toluidine blue, oil red o and alizarin red staining. The protein expression of NSE, LPL, collagen Ⅰ and collagen Ⅱ were examined by Western Blot. Results MTT assay showed that 1 g/L APS promoted the proliferation better than other concentrations especially in 48 hours. The morphologic change of the cell from BMSCs was uniformly positive to toluidine blue staining, and was negative to oil red o and alizarin red staining. Western blot showed that the protein expression of the cell from BMSCs was positive for NSE but negative for LPL, collagen Ⅰ and collagen Ⅱ. Conclusion BMSCs induced by APS can differentiate to neurone and fail to differentiate to adipocytes, osteoblasts and chondrocytes in vitro.