半夏总生物碱对6-OHDA诱导PC12细胞损伤的保护作用及机制

目的探讨半夏总生物碱(TAPT)对6羟基多巴胺(6-OHDA)诱导的大鼠肾上腺嗜铬细胞瘤细胞PC12细胞株损伤的保护作用及其机制。方法通过不同剂量TAPT预处理PC12细胞后,加入6-OHDA诱导氧化应激损伤。采用甲基噻唑基四唑比色法(MTT法)检测PC12细胞活力、天冬氨酸特异性半胱氨酸蛋白酶3(caspase-3)活性检测试剂盒测定caspase-3活性、黄嘌呤氧化酶法检测细胞总超氧化物歧化酶(T-SOD)活力,硫代巴比妥法检测丙二醛(MDA)的含量、按Fenton反应原理检测抑制羟自由基(·OH)的能力。结果浓度为100μmol/L的6-OHDA作用PC12细胞24h后,与正常对照组相比,细胞活力明显降低(P<0.05),T-SOD活性和抑制·OH的能力明显下降(P<0.05),而MDA含量和caspase-3活性明显升高(P<0.05);与6-OHDA组相比,TAPT处理组的PC12细胞活力逐渐升高(P<0.05),T-SOD活性和抑制·OH的能力明显上升(P<0.05),而MDA含量和caspase-3活性明显降低(P<0.05)。结论 TAPT对6-OHDA诱导的PC12细胞损伤具有一定的保护作用;提高T-SOD活性和抑制·OH的能力及降低MDA含量和caspase-3活性可能是其作用机制之一。

Protective effect of total alkaloids derived from Pinellia Ternate on PC12 cells injured by 6-hydroxydopamine and its mechanism

Objective To explore the protective effect of total alkaloids derived from Pinellia Ternate (TAPT) on pheochromocytoma cells (PC12 cells) injured by 6-hydroxydopamin (6-OHDA) and its mechanism.Methods The oxdative stress damage to PC12 cells who were pretreated with different doses of TAPT or not was induced by 6-OHDA.The viability,total superoxide dismutase (T-SOD) activity,the content of malondialdehyde (MDA),capability to inhibt hydroxyl free radical (·OH),and cysteinecontaining aspartate-specific proteases 3 (caspase-3) activity in the PC 12 cells were determined.Results The viability,T-SOD activity and capability to inhibit·OH were significantly lower,and the content of MDA and caspase-3 activity were significantly higher in the PC12 cells 24 hours after the treatment with 100μmol/L 6-OHDA than those in PC12 cells of the normal group (P<0.05).The cell viability,T-SOD activity and capability to inhibit·OH were significantly higher,and the content of MDA and caspase-3 activity were significantly lower in the PC12 cells pretreated by TAPT of different doses than those in PC12 cells of the model (control) group (P<0.05).Conclusion It is suggested that TAPT may inhibit 6-OHDA-induced damage to PC12 cells via the increases in T-SOD activity and capability to inhibit·OH and the decreases in the MDA content and caspase-3 activity.