YC-1对Aβ寡聚体毒性的保护作用及其相关机制

Protective Effect of YC-1 Against ADDLs-induced Toxicity and HIF-1α Related Mechanism

目的:探讨YC-1针对Aβ寡聚体(ADDLs)毒性的保护作用及其相关机制。方法:原代培养小鼠皮质神经元及星形胶质细胞混合培养体系,应用不同浓度ADDLs(500 nmol/L、1μmol/L、2μmol/L、5μmol/L、10μmol/L、20μmol/L)干预以模拟体外AD模型,应用YC-1对此模型进行干预,以CCK-8法检测细胞活力,Western blot检测NICD及缺氧诱导因子-1α(HIF-1α)水平,采用real-time PCR检测Caspase-3 mRNA水平。结果:浓度≥2μmol/L的ADDLs干预明显降低混合培养体系细胞活力水平,并呈现浓度依赖性(P<0.01)。10μmol/L ADDLs持续干预4 h显著降低细胞活力(P<0.01),YC-1可明显拮抗这种毒性作用(P<0.01)。10μmol/L ADDLs持续干预4 h明显上调NICD及HIF-1α水平(P<0.01),YC-1可拮抗这种提高(P<0.05或0.01)。10μmol/L ADDLs干预4 h显著提高Caspase-3的mRNA水平(P<0.01),YC-1可显著抑制这种上调(P<0.05)。结论:ADDLs表现出对原代培养小鼠皮质神经元及星形胶质细胞混合培养体系的毒性作用,YC-1可发挥保护作用。

ObjectiveTo study the protective effect of YC-1 against ADDLs （Aβ oligomers）-induced toxicity and related mechanism. Methods： Different concentrations （500 nmol/L, 1 μmol/L, 2 μmol/L, 5 μmol/L, 10 μmol/L, 20 μmol/L） of ADDLs were used to intervene the primary co-culture system of mouse cortical neurons and astrocytes, and YC-1 was applied at the same time. CCK-8 assays were used to assess cell vitality of co-culture cells. NICD and HIF-1α levels of primary co-culture system were assessed with western blot, and real-time PCR was used to detect apoptosis associated Caspase-3 mRNA level of co-culture system. Results：ADDLs at ≥2 μmol/L reduced the cell viability of the co-culture system in a concentration-dependent manner （ P〈0.01）. Exposure to 10 μmol/L ADDLs for 4 hours showed obvious toxic effect on the co-culture system （ P〈0.01）, while the application of YC-1 significantly attenuated the toxic effect induced by ADDLs （ 〈0.01）. The NICD and HIF-1α levels were up-regulated after the exposure of 10 μmol/L ADDLs for 4 hours （ P〈0.01）, while the application of YC-1 significantly attenuated the up-regulation of the protein levels （ P〈0.05 or 0.01）. The Caspase-3 mRNA level was increased after intervened by 10 μmol/L ADDLs for 4 hours （ P〈0.01）, and YC-1 significantly inhibited the increase （ P〈0.05）. Conclusion： The toxic effect induced by ADDLs on primary co-culture system of mouse cortical neurons and astrocytes could be protected by YC-1.