Protective effect on tanshinone II A on neural progenitor cell line C17.2

Aim:To investigate the protective effect of Tanshinone II A on neural progenitor cell line C17.2, and explore its possible mechanism. Methods:The experiment was performed in the Central Laboratory of Guangzhou Tissue Typing Center in Guangzhou Blood Center from 2005. Dr. David Walsh from the Department of Anatomy, The University of New South Wales, Australia, provided the C17.2 cell line. The mouse neural progenitor cell line C17.2 was cultured at the density of 1 ×109 L-1 in lMDM supplemented with 10% (v/v) fetal calf serum (FCS) in an atmosphere of 0.05 volume fraction CO2/saturated humidified air at 37°C. Nearly confluent C17.2 cells were digested with pancreatic enzyme containing 0.1 mmol/ LEDTA a room temperature and passaged at the ratio of 1:3. C17.2 cells at the density of 5x107 L-1 were seeded in a 96-well plate or 25 CM2 cultured flask. After being cultured in lMDM with 10% CS overnight, the cells were incubated with AAPH at 4 g/ L in serum-free lMDM to establish apoptosis models. C17.2 cells at the density of 5x107 L-1 were seeded in a 96-well plate. After being cultured in IMDM with 10% FCS overnight, the cells were incubated with AAPH at 4 g/L in serum-free lMDM. Control group did not receive tanshinone II A. Experiment group received tanshinone 1[ A at different doses (0.02, 0.05, 0.1 and 0.2 mg/L) for 8 hours. Cell activity was measured with MTT method: relative value of cell activity was equal to absorbance (A) of experimental group/A of control group ×100%. Cell apoptosis was measured with flow cytometry. Results: (1)After treatment with AAPH for 8 hours, which induced oxidative damages to C17.2 cells, most of the cells lost their normal fibroblast-like morphology and appeared round in shape. Many of them were detached. After being treated with tanshinone II A, cells kept normal mostly, and few appeared round. (2)Number of C17.2 cells in lMDM was 2.5-3 times than that in AAPH at 4 g/L in serum-free IMDM. Tanshinone II A at the concentrations of 0.02,0.05 and 0.1 mg/L had protective effects on C 17.2 cells. Tanshinone II A at over 0.2 mg/L concentration had lower protective effect on C17.2 cells. (3)The majority of C17.2 cells had intact mitochondria before AAPH, while few had early apoptotic and apoptotic cells. The treatment with AAPH increased the total apoptotic cells. The tanshinone II A significantly decreased early apoptosis. Conclusion: The tanshinone II A in vitro has anti-apoptosis effect on neural cells and can protect neural cells.