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伴侣自吞噬在MPP+损伤的SH-SY5Y细胞中的影响及葛根素的干预作用

Keywords: 葛根素,伴侣自吞噬,MPP+,SH-SY5Y细胞,ROS

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Abstract:

目的:研究葛根素通过分子伴侣自吞噬途径保护MPP+损伤的SH-SY5Y细胞。方法:以1mmol·L-1MPP+损伤SH-SY5Y细胞建立帕金森病细胞模型。采用CCK-8染色检测不同浓度的葛根素对MPP+损伤的SH-SY5Y细胞存活率的影响;透射电镜、AO染色观察自噬体的形成及检测溶酶体活性的变化;RT-PCR检测Lamp2a,Hsc70mRNA表达的变化,Westernblotting法检测细胞内Lamp2a,Hsc70,α-synuclein蛋白的表达。结果:在12.5~50.0μmol·L-1,预先30min加入葛根素可以保护SH-SY5Y细胞抵抗MPP+诱导的损伤,并呈一定的量效关系;AO染色和电子显微镜检测25.0,50.0μmol·L-1葛根素作用于1mmol·L-1MPP+损伤的SH-SY5Y细胞,细胞内出现自噬体,并且随着葛根素剂量的增加,胞质中形成的自噬体增多;流式细胞术检测结果显示50.0μmol·L-1葛根素可以拮抗1mmol·L-1MPP+诱导的SH-SY5Y细胞内ROS水平的增加,阻止氧化损伤的发生;RT-PCR和Westernblotting结果表明在12.5~50.0μmol·L-1葛根素通过上调细胞内Hsc70,Lamp2amRNA和蛋白水平保护MPP+造成的SH-SY5Y细胞损伤并抑制MPP+诱导的SH-SY5Y细胞α-synuclein蛋白的积聚。结论:葛根素可以拮抗MPP+诱导的SH-SY5Y细胞损伤,其保护机制可能与干预分子伴侣自吞噬途径有关。

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