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目的:探讨龟鹿二仙胶汤含药血清对骨髓间充质干细胞(BMSCs)向软骨细胞分化的干预作用。方法:抽取兔骨髓液,经体外分离、纯化、培养获得兔BMSCs,将第2代兔BMSCs分为龟鹿二仙胶汤中药含药血清组、软骨细胞组、单纯诱导剂组及空白对照组,进行培养1、2、3周后,通过细胞染色、透射电子显微镜、免疫组化等实验技术,观察各组对BMSCs诱导成软骨细胞的影响。结果:在转化生长因子-β(3TGF-β3)等诱导下龟鹿二仙胶汤中药血清可以明显促进软骨表型化BMSCs的GAG分泌及Ⅱ型胶原mRNA的表达,与软骨细胞组和单纯诱导剂组有显著性差异(P<0.01)。结论:龟鹿二仙胶汤含药血清可促进BMSCs向软骨细胞转化,优化骨组织工程的种子细胞体系,改善软骨组织的质量,延缓软骨的退化过程。
Objective: To investigate the effect of Guilu Erxian Decoction serum on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into chondrocytes. METHODS: Rabbit bone marrow fluid was collected and isolated, purified and cultured in vitro to obtain rabbit BMSCs. The second generation of rabbit BMSCs was divided into Guilu Erxian Decoction traditional Chinese medicine containing serum group, chondrocyte group, simple inducer group and blank control group. After 1, 2, and 3 weeks of culture, cell staining, transmission electron microscopy, and immunohistochemistry were used to observe the effect of each group on chondrocytes induced by BMSCs. Results: The serum of Guilu Erxianjiao decoction induced by transforming growth factor-β(3TGF-β3) could obviously promote GAG secretion and mRNA expression of type II collagen in cartilage phenotyped BMSCs, as well as chondrocyte group and simple induction. There was a significant difference in the agent group (P<0.01). Conclusion: The serum containing Guilu Erxian Decoction can promote the transformation of BMSCs into chondrocytes, optimize the seed cell system of bone tissue engineering, improve the quality of cartilage tissue, and delay the degeneration of cartilage.