丹参酮ⅡA对兔纤维环细胞能量代谢的保护作用(英文)

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

[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/mg-prot)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 （ TS H A） 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 TS Ⅱ A and IL - 1β was added to the medium for intervention ： no drug was added in group A as normal control, 4 μg/ml TS H A in group B, 10 μg/ml IL - 1β in group C, and both 10 μg/ml IL - 1β and different concentrations of TS H A 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）, MTr assay for cell proliferation, and Annexin V - 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 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 lower than that of group C （43. 11 ± 2. 7, P 〈 0. 01 and 11.71 ± 0. 32, P 〈 0. 01 ） . [ Conclusion ]TS Ⅱ A is able to promote cell proliferation and decrease cell apoptosis of AF by alleviating IL - 1β induced inhibition on cell energy metabolism.