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目的探究姜黄素(Cur)对鱼藤酮(Ro)诱导的PC12细胞损伤的保护作用及机制。方法建立鱼藤酮诱导的PC12细胞的帕金森病模型,利用姜黄素进行干预。实验分组为空白对照组,鱼藤酮模型组(终浓度:0.1μmol/L),姜黄素干预组(终浓度:0.5μmol/L、1.0μmol/L、5.0μmol/L、10μmol/L)。MTT比色法检测细胞活力,AO/EB荧光染色法观察细胞的凋亡情况,AnnexinⅤ-FITC/P双染检测细胞凋亡,Western-blot法检测细胞内IGF-1、Akt、FoxO3a的蛋白表达。结果 MTT结果显示:鱼藤酮组较对照组细胞活力明显降低,差异具有统计学意义(P<0.01),0.5μmol/L和1.0μmol/L姜黄素干预组可减轻0.1μmol/L鱼藤酮对PC12细胞增殖活力的影响,明显抑制了鱼藤酮对PC12细胞凋亡的诱导作用,与鱼藤酮组比较差异有统计学意义(P<0.01)Western-blot结果示:鱼藤酮组较对照组明显降低,差异具有统计学意义(P<0.01),0.5μmol/L和1.0μmol/L姜黄素干预组IGF-1、磷酸化的Akt(p-Akt)、磷酸化的FoxO3a(p-FoxO3a)表达与鱼藤酮组比明显升高,差异均有统计学意义(均P<0.01)。以上结果均显示5.0μmol/L和10μmol/L姜黄素干预组与鱼藤酮组比较差异无统计学意义(P>0.05);结论适宜浓度的姜黄素对鱼藤酮诱导PC12细胞损伤的PD模型具有保护作用,其保护作用呈浓度依赖性,其保护作用的机制可能与激活IGF-1/Akt/FoxO3a通路有关。
Objective To explore the protective effect and mechanism of Cur on the injury of PC12 cells induced by rotenone (Ro). Methods Parkinson ’s disease model of PC12 cells induced by rotenone was established and curcumin was used for intervention. The experimental group was divided into blank control group, rotenone model group (final concentration: 0.1μmol / L) and curcumin intervention group (final concentration: 0.5μmol / L, 1.0μmol / L, 5.0μmol / L, 10μmol / L). Cell viability was detected by MTT colorimetric assay. Cell apoptosis was observed by AO / EB staining. Apoptosis was detected by AnnexinⅤ-FITC / P double staining. The protein expression of IGF-1, Akt and FoxO3a was detected by Western-blot . Results The results of MTT showed that the viability of rotenone group was significantly lower than that of control group (P <0.01), and the effects of 0.5 μmol / L and 1.0 μmol / L curcumin on the proliferation of PC12 cells were inhibited by 0.1 μmol / L rotenone (P <0.01) Western-blot results showed: rotenone group was significantly lower than the control group, the difference was statistically significant (P <0.01), the effect of rotenone on the apoptosis of PC12 cells was significantly inhibited (P <0.01). The expressions of IGF-1, phosphorylated Akt (p-Akt), phosphorylated FoxO3a (p-FoxO3a) and rotenone group were significantly higher in the groups of 0.5μmol / L and 1.0μmol / , The differences were statistically significant (all P <0.01). The above results showed that there was no significant difference between 5.0μmol / L and 10μmol / L curcumin intervention groups and rotenone group (P0.05) .Conclusion Appropriate concentrations of curcumin have a protective effect on rotenone-induced PD model of PC12 cells, Its protective effect is concentration-dependent, and its protective mechanism may be related to the activation of IGF-1 / Akt / FoxO3a pathway.