姜黄素对ActD/TNF-α协同诱导PC12细胞损伤的保护作用

Protective effect of curcumin on ActD/TNF-α-induced synergistically insults in PC12 cells

目的:观察放线菌素D(ActD)和肿瘤坏死因子α(TNF-α)对大鼠肾上腺嗜铬细胞(PC12细胞)的协同损伤作用,探讨中药单体姜黄素对这种损伤的保护作用及机制。方法:采用MTT法确定实验药物的最佳浓度;LDH法检测PC12细胞的损伤;倒置显微镜观察细胞形态变化;caspase-3活性检测试剂盒检测caspase-3的活性。结果:ActD/TNF-α可致PC12细胞的生存力下降(P<0.05);细胞培养液中LDH的活性升高(P<0.05);PC12数量减少,体积缩小,突起变短;可致PC12细胞内caspase-3的活化增强(P<0.05)。浓度为5μmol/L姜黄素可阻止ActD/TNF-α所致的细胞的生存力下降(P<0.05);抑制ActD/TNF-α引起的细胞培养液中LDH的活性升高(P<0.05);拮抗ActD/TNF-α引起的细胞数量减少,体积变小,突起减少变短;抑制ActD/TNF-α引起的PC12细胞内caspase-3的活化(P<0.05)。结论:姜黄素可以拮抗ActD/TNF-α协同作用引起的PC12细胞损伤,其机制可能与抑制caspase-3的活化有关。

Aim：To observe the synergistic insults of ActD and TNF-ot on PC12 cells, and explore the protective effect of curcumin on the insults as well as its mechanism. Methods： MTT method was used to evaluate the optimal concentration of drugs, lactate dehydrogenase （LDH） assay to detect insult of PC12 cells in each group, inverted microscope to observe morphological change of PC12 ceils, caspase-3 activi- ty detection kit to detect the activity of caspase-3. Results： ActD/TNF-ot can reduce viability of PC12 ceils ; increase LDH activity in cell cultures ; induce reduction of cell number, shrink of cell volume, de- ducting and shortening of neurite; and increase activation of caspase-3 in PC12 cells, curcumin with a concentration of 5 I^mol/L can resist viability decrease of PC12 cells induced by ActD/TNF-ct （P 〈 0..05 ） ; inhibit the increase of LDH activity in cell cultures （ P 〈 0. 05 ） ; inhibit reduction of cell num- ber, shrink of cell volume, deducting and shortening of neurite; and inhibit the increase of caspase-3 ac- tivation in PC12 cells （P 〈0. 05）. Colzehzsion：Cureumin can resist the insult of PC12 cells which de- rived from the synergism of ActD/TNF-et, the mechanisms of which may be involved in its inhibition to the activation of caspase-3.