人参皂苷Rb1通过调控Nrf2/ARE信号通路抗氧糖剥夺/复氧后神经细胞焦亡的研究

Ginsenoside Rb1 inhibits neuronal pyroptosis after oxygen-glucose deprivation/reoxygenation via Nrf2/ARE signaling pathway

目的从Nrf2/ARE抗氧化信号通路探讨人参皂苷Rb1抗氧糖剥夺/复氧(OGD/R)后SH-SY5Y细胞焦亡的作用机制。方法野生型SH-SY5Y细胞设立对照组、模型组、不同剂量人参皂苷Rb1组,CCK-8法检测细胞存活率。构建Nrf2沉默SH-SY5Y(si-Nrf2-SH-SY5Y)细胞,设立对照组、模型组、人参皂苷Rb1组、si-Nrf2-SH-SY5Y模型组、si-Nrf2-SH-SY5Y人参皂苷Rb1组。CCK-8法检测细胞存活率;LDH漏出率、Hoechst/PI染色和caspase-1蛋白的表达评价细胞焦亡;免疫荧光染色检测Nrf2蛋白活化;Western blot法检测全细胞Nrf2和HO-1蛋白的表达。结果人参皂苷Rb1能够提高OGD/R后细胞存活率,降低细胞焦亡,上调Nrf2蛋白的表达,维持HO-1蛋白的高表达。沉默Nrf2基因可导致细胞损伤和细胞焦亡进一步加重,同时人参皂苷Rb1激活Nrf2/ARE信号通路和抗细胞损伤、细胞焦亡的作用受到抑制。结论Nrf2/ARE抗氧化信号通路负调控OGD/R后SH-SY5Y细胞焦亡,人参皂苷Rb1通过调控Nrf2/ARE抗氧化信号通路,从而发挥抗细胞焦亡的作用。

Aim To investigate the mechanism of ginsenoside Rb1 inhibiting SH-SY5Y cell pyroptosis after oxygen-glucose deprivation/reperfusion from Nrf2/ARE antioxidant signaling pathway.Methods Wild-type SH-SY5Y cells were set up in control group,model group,and different doses of ginsenoside Rb1 group.Cell survival rate was detected by CCK-8 method.Nrf2-silenced SH-SY5Y(si-Nrf2-SH-SY5Y)cells were constructed.Control group,model group,ginsenoside Rb1 group,si-Nrf2-SH-SY5Y model group and si-Nrf2-SH-SY5Y ginsenoside Rb1 group were established.CCK-8 was used to detect cell survival rate;LDH leakage rate,Hoechst/PI staining and the expression of caspase-1 protein were used to evaluate pyroptosis.The expression of activated Nrf2 protein was detected by immunofluorescence.The expression of whole cell Nrf2 and HO-1 protein were detected by Western blot.Results Ginsenoside Rb1 could improve cellsurvival rate and decrease pyroptosis after OGD/R.Ginsenoside Rb1 could up-regulate the expression of Nrf2 protein and maintain the high expression of HO-1 protein.Silencing Nrf2 gene could lead to further aggravation of cell injury and pyroptosis;meanwhile,the effects of ginsenoside Rb1 activating Nrf2/ARE signaling pathway and inhibiting cell injury and pyroptosis were inhibited.Conclusions Nrf2/ARE antioxidant signaling pathway can negatively regulate SH-SY5Y cell pyroptosis after OGD/R,ginsenoside Rb1 can play an role of inhibiting pyroptosis by regulating Nrf2/ARE antioxidant signaling pathway.