The effects of andrographolide on human mesenchymal stem cells derived from placenta

This study aims to investigate the effect of Andrographolide on mesenchymal stem cell derived from placenta (PL-hMSCs). PL-hMSCs were isolated from placental tissues and were characterized before using. Firstly, the effect of Andrographolide on cell viability was performed by MTT assay after incubation with 0.01-50 µM of Andrographolide for 24-120 h. Cell proliferation were examined by BrdU assay after incubation with 0.01-20 µM of Andrographolide for 24-120 h. Osteoblast differentiation was determined by alkaline phosphatase (ALP) staining and activity. Bone mineralization was investigated by alizarin red staining assay. Real time PCR was performed to examine the effect of Andrographolide on the expression of osteoblast-specific genes including Runt-related transcription factor 2 (Runx2), Osterix (OSX), Osteocalcin (OCN). The effect of Andrographolide on mRNA expression of the key proteins of bone remodeling which Osteoprotegerin (OPG) and Receptor activator of nuclear factor-κB ligand (RANKL) were examined by qRT-PCR. Andrographolide at 0.01-10 µM had no toxic to PL-hMSCs. PL-hMSCs proliferation were increased after treatment with Andrographolide at 0.1-5 µM for 24-120 h. Moreover, Andrographolide at 0.1-5 µM also accelerated osteogenic differentiation of PL-hMSCs, as indicated by the increases in ALP expression, ALP activity and osteoblast-specific mRNA expression including Runx2, OSX and OCN. Interestingly, Andrographolide promoted bone mineralization, as indicated by the increases in calcium deposition in osteoblast cells. In conclusion, Andrographolide promotes PL-hMSCs proliferation, osteogenic differentiation, osteoblast function. Andrographolide promote bone formation of PL-hMSCs on OPG and RANKL pathway is one of the mechanism of action of the compound. Base on these available data, Andrographolide may have potential for development as a new alternative agent using for therapeutic application of bone related disease.