丹参酮ⅡA对兔纤维环细胞能量代谢的保护作用

目的]考察丹参酮ⅡA(TSⅡA)对白细胞介素-1β(IL-1β)诱导的兔纤维环细胞能量代谢障碍的保护作用.方法]藻酸盐串珠立体培养兔纤维环细胞,将细胞分为7组,在培养过程中加入不同浓度的药物:A组为空白对照不加入药物,B组加入4 μg/ml TSⅡA,C组加入10ng/ml IL-1β,D～G组在给予10 ng/mlIL-1β同时分别加入0.5、1、2和4 μg/ml TSⅡA.于培养3 d后行Na+-K+-ATP酶活性检测、,琥珀酸脱氢酶活性检测、MTT法细胞增殖情况检测以及细胞凋亡的流式细胞仪检测.结果]G组Na+-K+-ATP酶活性(3.23±0.28U/mgprot)较C组(1.118±0.15U/mgprot)明显增高(P<0.01),与A组接近(3.57±0.15 U/mgprot)(P>0.05).G组琥珀酸脱氢酶活性(12.48±0.97U/mgprot)较c组(3.03±0.60 U/mgprot)明显增高(P<0.01),与A组接近(14.24±1.56 U/mgprot)(P>0.05).G组MTT试验吸光度(0.77±0.06)较C组(0.31±0.07)明显增高(P＜0.01),随着TSⅡA浓度的升高,D～G组吸光度随着TsIIA上升而上升.G组细胞死亡细胞比例和凋亡细胞比例分别为21.08±1.46%和8.99±0.33%,均显著低c组(43.11±2.7,P<0.01和11.71±0.32,P<0.01).结论]TSIIA能够减轻IL-1β对纤维环细胞能量代谢的抑制作用,从而改善纤维环细胞的增殖、死亡及凋亡.

Protective effects of Tanshinone IIA against interleukin-1 beta induced obstruction of energy metabolism of rabbit annulus fibrosus cells in vitro

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.