透骨消痛颗粒含药血清诱导骨髓基质干细胞向软骨细胞分化

Tougu Xiaotong Granule containing serum induces cartilage differentiation of bone marrow stromal stem cells

背景:骨髓基质干细胞根据环境而有不同的分化路径,在特定的环境中有分化为透明软骨细胞的趋势,可能为治疗骨性关节炎提供新思路。目的:观察透骨消痛颗粒含药血清对骨髓基质干细胞向软骨分化的影响。方法:取SD大鼠四肢建立体外培养的骨髓基质干细胞,取第3代骨髓基质干细胞进行干预,分为生理盐水血清组、透骨消痛颗粒水提物血清组、透骨消痛颗粒醇提物血清组、诱导软骨剂组、透骨消痛颗粒水提物血清+诱导软骨剂组、透骨消痛颗粒醇提物血清+诱导软骨剂组。Real time PCR及Western Blot检测Sox9、collagenⅡ、collagenⅩ mRNA及蛋白表达。结果与结论:含药血清干预14d后,Sox9、collagenⅡ、collagenⅩ mRNA及蛋白表达水平,透骨消痛颗粒水提物血清组、透骨消痛颗粒醇提物血清组、诱导软骨组、透骨消痛颗粒水提物血清+诱导软骨剂组、透骨消痛颗粒醇提物血清+诱导软骨剂组明显高于生理盐水血清组(P<0.05,P<0.01),诱导软骨组、透骨消痛颗粒水提物血清+诱导软骨剂组、透骨消痛颗粒醇提物血清+诱导软骨剂组明显高于透骨消痛颗粒水提物血清组、透骨消痛颗粒醇提物血清组(P<0.01),其中Sox9表达透骨消痛颗粒水提物血清+诱导软骨剂组高于诱导软骨。说明透骨消痛颗粒含药血清通过上调Sox9的表达,加速骨髓基质干细胞细胞向软骨细胞分化。

BACKGROUND： Bone marrow stromal stem cells have multilineage differentiation capacity and can differentiate into transparent chondrocytes under certain conditions, which can provide new thoughts for treatment of osteoarthritis. OBJECTIVE： To investigate the effects of Tougu Xiaotong Granule containing serum on the cartilage differentiation of bone marrow stromal stem cells. METHODS： Bone marrow stromal stem cells from Sprague-Dawley rat limbs were cultured in vitro, and those cells at passage 3 were used in the study. Cells were divided into six groups： saline serum group, Tougu Xiaotong Granule water extract group, Tougu Xiaotong Granule alcohol extract group, chondroinductive group, Tougu Xiaotong Granule water extract and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group. The Sox9, collagen Ⅱ, and collagen X mRNA and protein expression levels were tested by real-time polymerase chain reaction and western blot analysis. RESULTS AND CONCLUSION： After cells were intervened with drug-containing serum for 14 days, the Sox9, collagen Ⅱ, and collagen X mRNA and protein expression in Tougu Xiaotong Granule water extract group,Tougu Xiaotong Granule alcohol extract group, chondroinductive group, Tougu Xiaotong Granule water extract and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group were significantly higher than that in saline serum group （P〈0.05, P 0.01）. In addition, the chondroinductive group, Tougu Xiaotong Granule water extract serum and chondroinductive group, Tougu Xiaotong Granule alcohol extract and chondroinductive group showed significantly higher expression levels than Tougu Xiaotong Granule water extract serum group, Tougu Xiaotong Granule alcohol extract group （P 0.01）. Sox9 expression in Tougu Xiaotong Granule water extract serum and chondroinductive group were significantly higher than that in the chondroinductive group. Experimental findings indicate that, Tougu Xiaotong Granule containing serum can accelerate bone marrow stromal stem cells differentiate into cartilage cells by up-regulation of Sox9 expression.