红花黄色素对Aβ_(1-42)诱导的SH-SY5Y细胞损伤的保护作用研究

Study on the protective effects of Safflower Yellow on injury of cultured SH-SY5Y cells induced by Aβ_(1-42)

目的:探讨红花黄色素对Aβ_(1-42)诱导的SH-SY5Y细胞损伤的保护作用及其机制。方法:采用Aβ_(1-42)损伤SH-SY5Y细胞模拟体外AD,不同浓度(0. 01、0. 1、1g/l)红花黄色素进行干预。倒置显微镜下观察细胞形态; MTT法测定细胞存活率;酶联免疫吸附法(ELISA)检测细胞内Aβ_(1-42)水平; Western Blot检测SH-SY5Y细胞内APP、BACE-1、SAPPα、NEP、IDE、ERK和P-ERK蛋白表达水平。结果:红花黄色素各剂量均能够减少Aβ_(1-42)引起的SH-SY5Y细胞的损伤,使细胞的存活率显著提高,且具有剂量依赖性;ELISA结果显示,红花黄色素0. 1、1g/L可降低SH-SY5Y细胞内的Aβ_(1-42)水平; Western Blot结果显示红花黄色素各剂量均可以上调SAPPα、NEP、IDE和P-ERK蛋白的表达,下调APP和BACE-1蛋白的表达。结论:红花黄色素可以通过调节Aβ_(1-42)相关代谢酶而减少Aβ_(1-42)的含量,发挥抗AD作用,该作用可能与MAPK/ERK通路的激活有关。

Objective： To investigate the protective effects and mechanisms of safflower yellow on damage of cultured SH-SY5 Y cells induced by Aβ1-42. Method： SH-SY5 Y cells were cultured in vitro,with contained Aβ1-42 for preparing AD models and different concentrations of safflower yellow（0. 01,0. 1,1 g/l） were used for intervention. Morphological picture of cultured cells were observed by inverted microscope and cell viability was assayed by tetramethyl triazole salt（MTT）. The contents of secreted Aβ1-42 levels were measured by ELISA kit. The APP,BACE-1,SAPPα,NEP,IDE,ERK and P-ERK protein levels were detected by Western blot. Result： Safflower yellow relieved the damage of SH-SY5 Y cells induced by Aβ1-42 and significantly increased the cell survival rate in a dose-dependent manner. The results of ELISA showed that safflower yellow could decrease Aβ1-42 levels in cells. The results of Western blot showed that safflower yellow increased the protein expression of SAPPα,NEP,IDE,ERK and decreased APP and BACE-1 levels. Conclusion： These observations suggest that safflower yellow exerts its effect on anti-AD by regulating Aβ1-42-related metabolic enzymes to reduce the content of Aβ1-42,which may be related to the activation of MAPK/ERK signaling pathway.