Sestrin 2通过Nrf2/xCT途径减轻七氟醚诱导的HT22细胞铁死亡研究

Sestrin 2 extenuates neuronal ferroptosis induced by sevoflurane through the Nrf2/xCT pathway

目的探讨Sestrin 2(SESN2)在七氟醚(SEV)诱导神经元损伤的作用及其分子调控机制。方法将HT22小鼠海马神经元细胞用0%、1%、2%、4%七氟醚混合气体和/或铁死亡抑制剂Ferrostatin-1(FER-1)处理6 h。将SESN2过表达载体(SESN2-OE)和相应的对照(NC)转染至HT22细胞中。检测细胞活力及铁离子、活性氧(ROS)、丙二醛(MDA)和总谷胱甘肽(GSH)水平。实时定量PCR(qPCR)和Western blotting测定mRNA和蛋白水平。结果 SEV明显抑制SESN2 mRNA和蛋白的表达(t值分别为22.904、37.432,均P<0.05),转染SESN2-OE后SESN2 mRNA和蛋白表达明显增加(t值分别为12.703、11.687,均P<0.01)。SEV处理使HT22细胞中铁离子、ROS和MDA的水平明显增加(t值分别为29.031、18.819、28.054,均P<0.05),而GSH、Nrf2和胱氨酸/谷氨酸逆向转运体(xCT)的水平明显降低(t值分别为14.617、34.513、13.836,均P<0.05)。转染SESN2-OE使SEV处理的HT22细胞中铁离子、ROS和MDA的水平明显降低(t值分别为8.342、9.373、7.381,均P<0.05),而GSH、核因子E2相关因子(Nrf2)和xCT的水平明显增加(t值分别为12.718、10.102、9.814,均P<0.05)。结论 Sestrin 2通过激活Nrf2/xCT途径抑制七氟醚诱导的HT22神经元铁死亡。

Objective To explore the role of Sestrin 2(SESN2) in neuronal ferroptosis induced by sevoflurane(SEV) and its possible mechanism. Methods Mouse hippocampal neuron cells HT22 were treated with 0%, 1%, 2% or 4% sevoflurane and iron death inhibitor Ferrostatin-1(FER-1) for 6 h. SESN2 overexpression vector(SESN2-OE) and corresponding control(NC) were transfected into HT22 cells. The CCK-8 assay was used in detecting cell viability, and the corresponding kit was used in detecting the levels of iron, reactive oxygen species(ROS), malondialdehyde(MDA) and glutathione(GSH). The expression levels of genes and proteins were detected through real-time quantitative PCR(qPCR) and western blot assay. Results qPCR and western blot results showed that SEV treatment significantly inhibited the expression of SESN2 mRNA and protein(t=22.904, 37.432;P<0.05), whereas transfection with SESN2-OE significantly increased the expression levels of SESN2 mRNA and protein in SEV-treated HT22 cells(t=12.703, 11.687;P<0.05). SEV treatment significantly increased the levels of iron, ROS and MDA in the HT22 cells(t=29.031, 18.819, 28.054) but significantly reduced the levels of GSH, Nrf2 and xCT(t=14.617, 34.513, 13.836;all P<0.05). Transfection with SESN2-OE significantly reduced the levels of iron, ROS and MDA in SEV-treated HT22 cells(t=8.342, 9.373, 7.381) but significantly increased the levels of GSH, Nrf2 and xCT(t=12.718, 10.102, 9.814;P<0.05). Conclusion Sestrin 2 inhibited sevoflurane-induced HT22 neuronal ferroptosis by activating the Nrf2/xCT pathway.