丹参酮ⅡA对兔纤维环细胞能量代谢的保护作用

Protective Effects of Tanshinone ⅡA on Energy Metabolism in Rabbit Annulus Fibrosus Cells

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4μg/mlTSⅡA,C组加入10ng/mlIL-1β,D～G组在给予10ng/mlIL-1β同时分别加入0.5、1、2和4μg/mlTSⅡA。于培养3d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±0.15U/mgprot)(P>0.05)。G组琥珀酸脱氢酶活性(12.48±0.97U/mgprot)较C组(3.03±0.60U/mgprot)明显增高(P<0.01),与A组接近(14.24±1.56U/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%,均显著低于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。

Objective：To investigate the protective effect of tanshinone IIA （TSⅡ A） against interleukin-1β （IL-1β） induced energy dysmetabolism in rabbit annulus fibrosus （AF） cells. Methods： Rabbit AF cells were cultured in 3 dimensional alginate beads and randomly divided into 7 groups： no drug for group A as normal control, 4μg/ml TSU A for group B, 10ng/ ml IL-1β for group C, both 10ng/ml IL-1β and different concentrations of TS Ⅱ A for group D-G （0. 5μg/ml, 1μg/ml, 2μg/ml or 4μg/ml, respectively）. After 3 days, the cells were collected to measure the activity of Na^＋-K^＋-ATPase and Suceino Dehydrogenase （SDH）. MTT assay were used to observe cell proliferation, and Annexin V-PI staining was performed to detect cell apoptosis. Results： The activity of Na^＋ -K^＋ ATPase of group G was increased significantly compared with group C （P〈0.01）, and was close to that of group A （P2〉0. 05）. SDH activity 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） and was close to that of group A （14.24±1.56） U/mgprot （P〉0.05）. The absorbance of MTT assay of group G was significantly increased compared with group C （P〈0.01）. The absorbance of group D-G increased as the concentrations of TS Ⅱ A 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 higher than that of group C （43.11±2.7, P〈0.01 and 11.71±0.32, P〈0.01）. Conclusion：TSU A is able to promote cell proliferation and decrease cell apoptosis of AF by alleviating IL-1β induced inhibition on cell energy metabolism.