Protective effects of Tanshinone IIA against interleukin-1 beta induced obstruction of energy metabolism of rabbit annulus fibrosus cells in vitro / 中国矫形外科杂志

ABSTRACT

[Objective]To investigate the protective effect of Tanshinone IIA (TSⅡA) against interleukin-1β (IL-1β) induced obstruction of energy metabolism of rabbit annulus fibrosus cell in vitro.[Methods]Rabbit annulus fibrosus (AF) cells were cultured in 3-dimension alginate beads and randomly divided into 7 groups. Various concentrations of TSⅡA and IL-1β was added to the medium for intervention no drug was added in group A as normal control, 4 μg/ml TSⅡA in group B, 10 μg/ml IL-1β in group C, and both 10 μg/ml IL-1β and different concentrations of TSⅡA in groups D-G (0.5 μg/ml, 1 μg/ml, 2 μg/ml and 4 μg/ml respectively). After 3 days of incubation, the cells were collected for measuring the activity of Na+-K+-ATPase and succinate dehydrogenase (SDH), MTT assay for cell proliferation, and AnnexinⅤ-PI staining for cell apoptosis.[Results]The activity of Na+-K+-ATPase of group G (10 μg/ml IL-1β+4μg/ml TS IIA; 3.23±0.28 U/mgprot) was increased significantly as compared with group C (10 μg/ml IL-1β; 1.118±0.15 U/mgprot, P＜0.01). The activity of SDH of group G was 12.48±0.97 U/mgprot, which was obviously higher than that of group C (3.03±0.60 U/mgprot, P＜0.01). The absorbance of MTT assay of group G (0.77±0.06) was significantly increased as compared with group C (0.31±0.07,P＜0.01). The absorbance of groups D-G increased as the concentration of TSⅡA increased. The apoptotic cell rate and dead cell rate of group G was 21.08±1.46% and 8.99±0.33%, which were both lower than that of group C (43.11±2.7,P＜0.01 and 11.71±0.32,P＜0.01).[Conclusion]TSⅡA is able to promote cell proliferation and decrease cell apoptosis of AF by alleviating IL-1β induced inhibition on cell energy metabolism.