Department of Orthopaedics, The First Affiliated Hospital, Shantou University Medical College, 57 Chang Ping Road, Shantou, Guangdong 515041, China.
J Ethnopharmacol. 2011 Mar 24;134(2):268-74. doi: 10.1016/j.jep.2010.11.075. Epub 2010 Dec 16.
Panax notoginseng saponins (PNS) is the main effective component of Panax notoginseng, have various pharmacologic activities such as antioxidant, anti-inflammatory, and estrogen-like bioactivities, have been shown to be an effective agent on anti-osteoporosis. Bone marrow stromal cells (BMSCs) play a crucial homeostatic role in skeletal modeling and remodeling due to their capability to differentiate into osteooblasts. Whether PNS has effect on osteogenic differentiation of BMSCs are unknown. This study was designed to investigate the effects of PNS on the proliferation and osteogenic differentiation of BMSCs in vitro.
When BMSCs cultivated in the basal medium or the osteogenic induction medium (OS with or without PNS), cell proliferation was analyzed using an MTT assay, the mineralization was assessed using Alizarin red S staining, the alkaline phosphatase activity was measured using a commercial kit, the mRNA level of osteogenic gene and PPARγ2 gene were determined using RT-PCR, the protein level of PPARγ2 was analyzed by Western blotting.
BMSCs cultured in the basal medium with PNS caused a significant increase in proliferation. PNS treatment increased ALP activity, Alizarin red S staining and mRNA level of ALP, Cbfa 1, OC, and BSP, whereas decreased the mRNA level and protein expression of PPARγ2 during osteogenic induction. In addition, the effects of PNS treatment were dose-dependent relationship.
PNS could stimulate BMSCs proliferation and promote their osteogenic differentiation by up-regulation expression of osteogenic marker gene and down-regulation expression of adipogenic marker gene in a dose-dependent manner. Thus, PNS may play an important therapeutic role in osteoporosis patients by improving osteogenic differentiation of BMSCs.
三七总皂苷(PNS)是三七的主要有效成分,具有抗氧化、抗炎和类雌激素等多种药理活性,已被证明是一种有效的抗骨质疏松药物。骨髓基质细胞(BMSCs)由于能够分化为成骨细胞,在骨骼建模和重塑中起着至关重要的动态平衡作用。PNS 是否对 BMSCs 的成骨分化有影响尚不清楚。本研究旨在探讨 PNS 对 BMSCs 体外增殖和成骨分化的影响。
当 BMSCs 在基础培养基或成骨诱导培养基(含或不含 PNS 的 OS)中培养时,使用 MTT 测定法分析细胞增殖,使用茜素红 S 染色评估矿化,使用商业试剂盒测定碱性磷酸酶活性,使用 RT-PCR 测定成骨基因和 PPARγ2 基因的 mRNA 水平,使用 Western blot 分析 PPARγ2 蛋白水平。
在含 PNS 的基础培养基中培养的 BMSCs 增殖明显增加。PNS 处理增加了 ALP 活性、茜素红 S 染色和 ALP、Cbfa1、OC 和 BSP 的 mRNA 水平,而在成骨诱导过程中降低了 PPARγ2 的 mRNA 水平和蛋白表达。此外,PNS 处理的效果呈剂量依赖性。
PNS 可通过上调成骨标志物基因的表达和下调脂肪标志物基因的表达,以剂量依赖的方式刺激 BMSCs 增殖并促进其成骨分化。因此,PNS 通过改善 BMSCs 的成骨分化,可能在骨质疏松症患者的治疗中发挥重要作用。