透骨消痛颗粒对BMSCs向软骨细胞分化的影响

Effects of Tougu Xiaotong Granule on Differentiation of BMSCs into Chondrocytes

目的探讨透骨消痛颗粒含药血清诱导骨髓间充质干细胞(BMSCs)向软骨细胞分化的分子生物学机制。方法从5只4周龄SD大鼠骨髓中分离出BMSCs,体外传代培养,取第3代BMSCs分为空白对照组、软骨诱导剂组及透骨消痛颗粒组,进行培养1、2周后,通过透射电镜、RT-PCR等实验技术,观察各组对BMSCs诱导成软骨细胞的影响。结果透射电镜观察可见软骨表型化的BMSCs多数为近椭圆形,表面突起多,细胞核大,粗面内质网丰富,内质网池扩张明显,高尔基体丰富。在TGF-β3等诱导因子作用下,透骨消痛颗粒含药血清可以促进Sox9 mRNA、Cbfa1mRNA在细胞分化过程中的表达,与软骨诱导剂组相比表达量有显著性差异(P<0.05)。结论透骨消痛颗粒含药血清可促进BMSCs向软骨细胞转化,改善软骨组织的质量,延缓软骨的退化。

Objective To explore the molecular biological mechanism of differentiatioi5 of bone marrow stromal cells （BMSCs） into chondrocytes induced by Tougu Xiaotong granule drug-containing serum. Methods BMSCs were isolated from bone marrow of five 4-week-old SD rats and passage cultured in vitro, and the third generation of BMSCs were divided into control group, cartilage inducer group and Tougu Xiaotong granule group, then after one or two weeks, the effects of each group on BMSCs, which were induced into chondrocytes, were observed by transmission electron microscope and RT-PCR. Results Most of BMSCs, which had been induced, were nearly ellipse, rich in rough endoplasmic reticulum and apparato reticulare, and had dense surface projections, large cell nucleus and inflated endoplasmie retieulum ci9tema. Under the induction of TGF-β3, Tougu Xiaotong granule drug-containing serum group could promote the expressions of Sox9 mRNA and Cbfal mRNA in the course of cell differentiation, and there was a significantly difference compared with the cartilage inducer group （P〈0.01）. Conclusion Tougu Xiaotong granule drug-containing serum can promote the differentiation of BMSCs into chondrocytes, improve the quality of cartilaginous tissue, and postpone the degeneration of cartilage.