PI3K/Akt信号通路在硫化氢保护PC12细胞对抗化学性缺氧损伤的作用

目的探讨硫化氢(H2S)对PC12细胞PI3K/Akt信号通路的影响及该通路在H2S神经保护中的作用。方法Western blot法检测H2S供体硫氢化钠(NaHS)处理PC12细胞诱导Akt磷酸化的水平;CCK-8比色法检测细胞存活率;应用碘化丙啶(PI)染色流式细胞术检测细胞凋亡率。结果应用不同浓度NaHS处理PC12细胞30 min,在50～400μmol.L-1浓度范围内,呈浓度依赖性地上调Akt磷酸化的水平,但随着NaHS浓度的增加,磷酸化Akt表达量逐渐下降;Ly294002明显抑制了NaHS对Akt磷酸化水平的上调作用。NaHS预处理可以保护PC12细胞对抗600μmol.L-1CoCl2诱导的损伤,使细胞存活率提高及细胞凋亡率降低。而在预处理前使用PI3K/Akt信号通路抑制剂Ly294002,则明显地减弱了H2S的神经细胞保护作用。结论 H2S可通过激活PI3K/Akt信号通路保护PC12细胞对抗化学性缺氧损伤。

Role of PI3K/Akt in protective effect of hydrogen sulfide against PC12 cells injuries induced by chemical hypoxia

Aim To explore the role of PI3K/Akt pathway in hydrogen sulfide-induced neuroprotection of PC12 cells.Methods The level of p-Akt of PC12 cells induced by H2S donor(NaHS)was evaluated by Western blot.Cell viability was tested by cell counter kit-8.Apoptotic rate was evaluated by propidium iodide(PI) staining and flow cytometry(FCM).Results At the concentrations from 50 to 400 μmol·L-1 NaHS for 30 min,p-Akt level was upregulated in a dose-dependent manner,peaking at 400 mol ·L-1 NaHS treatment.As the dose of NaHS increased,p-Akt expression decreased.H2S preconditioning markedly protected PC12 cells against injuries induced by CoCl2,increasing the viability of cells and decreasing the percentage of apoptotic cells.Ly294002,an inhibitor of PI3K,not only decreased p-Akt level but also obviously lowered the neuroprotection of H2S.Conclusion H2S can protect PC12 cells against chemical hypoxia injuries by activiating PI3K/Akt pathway.