基于铁死亡探究牛磺酸对鱼藤酮致神经细胞损伤的保护作用

STUDY ON THE PROTECTIVE EFFECTS OF TAURINE ON ROTENONE-INDUCED NEURONAL DAMAGE BASED ON FERROPTOSIS

目的采用鱼藤酮诱导SH-SY5Y细胞损伤建立帕金森病(Parkinson’s disease,PD)样病变细胞模型,基于铁死亡探讨牛磺酸的保护作用及机制。方法实验分为对照组、鱼藤酮组、鱼藤酮+牛磺酸组、牛磺酸组;对照组、鱼藤酮组、鱼藤酮+铁死亡抑制剂组、铁死亡抑制剂组。通过Western blot、MTT细胞毒性检测试剂盒、乳酸脱氢酶细胞毒性检测试剂盒检测SH-SY5Y细胞损伤状况;运用Westernblot和铁含量检测试剂盒检测铁死亡水平;运用总谷胱甘肽检测试剂盒、脂质氧化(MDA)检测试剂盒、活性氧检测试剂盒、总SOD活性检测试剂盒检测氧化应激水平;运用Westernblot检测Nrf2/Keap1/SLC7A11信号通路活化水平。结果结果显示,与对照组比较,鱼藤酮组酪氨酸羟化酶表达量降低、细胞存活率降低、细胞死亡率升高、铁死亡和氧化应激水平升高以及Nrf2/Keap1/SLC7A11通路活化水平降低,P<0.05,差异具有统计学意义;与鱼藤酮组比较,鱼藤酮+牛磺酸组酪氨酸羟化酶表达量增加、细胞存活率升高、细胞死亡率降低、铁死亡和氧化应激水平降低以及Nrf2/Keap1/SLC7A11通路活化水平升高,P<0.05,差异具有统计学意义;与鱼藤酮+牛磺酸组比较,牛磺酸组上述各项指标变化无显著差异。运用铁死亡抑制剂显著降低了鱼藤酮致SH-SY5Y细胞损伤和氧化应激水平。结论在鱼藤酮诱导PD样病变细胞模型中,牛磺酸可通过Nrf2/Keap1/SLC7A11信号通路抑制铁死亡及氧化应激,发挥神经保护作用。

Objective SH-SY5Y cell model with Parkinson’s disease(PD)-like lesions was induced by rotenone to explore the protective effects of taurine.Methods Western blot,MTT assay kit and lactate dehydrogenase cytotoxicity assay kit were used to detect the levels of cell damage.Western blot and iron assay kit were used to detect the levels of ferroptosis.Total glutathione assay kit,lipid oxidation assay kit,reactive oxygen species assay kit and SOD activity detection kit were used to detect the levels of oxidative stress.Western blot was used to detect the activation of Nrf2/Keap1/SLC7A11 pathway.Results Compared with the control group,the inactivation of tyrosine hydroxylase(TH)as well as the levels of cell death,ferroptosis and oxidative stress were increased while the activation of Nrf2/Keap1/SLC7A11 pathway was decreased in rotenone group(P<0.05).Compared with the rotenone group,the inactivation of TH as well as the levels of cell death,ferroptosis and oxidative stress were decreased while the activation of Nrf2/Keap1/SLC7A11 pathway was increased in rotenone+taurine group(P<0.05).Compared with the rotenone+taurine group,the above indicators did not change significantly in taurine group.In addition,the levels of cell death and oxidative stress were significantly reduced by ferroptosis inhibitors(Ferrostatin-1)in rotenone-treated SH-SY5Y cells.Conclusion Taurine mitigates oxidative stress mediated by ferroptosis via Nrf2/Keap1/SLC7A11 signaling pathway in rotenone-induced SH-SY5Y cell model with PD-like lesions.[ACTA NUTRIMENTA SINICA,2023,45(1):51-56,62]