[关键词]
[摘要]
目的 研究刺老苞根皮水提物对成骨细胞TGF-β/BMPs信号通路的影响。方法 建立大鼠骨折模型,随机分为模型组,刺老苞根皮水提物低、中、高剂量(3.6、1.8、0.9 g/kg)组,ig给药7 d后腹主动脉取血,分离血清;培养大鼠原代成骨细胞,经碱性磷酸酶染色、茜素红染色鉴定合格后,分别以对应组别的动物血清连续培养48 h,采用实时荧光定量PCR(qRT-PCR)和Western blotting技术分别检测各组细胞中骨形态发生蛋白质-2(BMP-2)、转化生长因子-β(TGF-β)、Smad家族相关蛋白1、2(Smad-1、Smad-2)mRNA和蛋白表达。结果 与模型组比较,刺老苞根皮水提物不同浓度的含药血清组BMP-2、TGF-β、Smad-1 mRNA表达均显著升高(P<0.05、0.01),中、高剂量含药血清组Smad-2 mRNA表达显著升高(P<0.05);低、中、高剂量含药血清组的BMP-2、TGF-β、Smad-1、Smad-2蛋白表达均显著升高(P<0.05)。结论 刺老苞根皮水提物通过上调TGF-β/BMPs信号传导通路中的BMP-2、TGF-β、Smad-1、Smad-2表达,促进成骨细胞增殖。
[Key word]
[Abstract]
Objective To study the effect of aqueous extract from Araliae Echinocaulis Radicis et Cortex (AEAE) on expression of TGF-β/BMPs signaling pathway in osteoblasts. Methods The fracture model of rat was established and randomly divided into model group, AEAE low, middle and high dose (3.6, 1.8, 0.9 g/kg) group. After ig administration for 7 d, the blood was taken from abdominal aorta and the serum was separated. The rats primary osteoblasts were cultured and qualified after alkaline phosphatase staining and alizarin red staining. The osteoblasts were cultured continuously for 48 h in animal serum of the corresponding group. The contents of BMP-2, TGF-β, Smad-1 and Smad-2 were detected by qRT-PCR and Western blotting method. Results Compared with model group, the mRNA level of BMP-2, TGF-β and Smad-1 in AEAE low, middle and high dose group, Smad-2 mRNA level in middle and high dose group, and the protein level of BMP-2, TGF-β, Smad-1 and Smad-2 in low, middle and high dose groupwere significantly higher than those in model group(P<0.05 and 0.01). Conclusion AEAE can promote the proliferation of osteoblasts by up-regulating the expression of BMP-2, TGF-β, Smad-1 and Smad-2 in TGF-β/BMPs signaling pathway.
[中图分类号]
[基金项目]
国家自然科学基金(81173361、81270051、81473451)