舒芬太尼对1-甲基-4-苯基-吡啶离子诱导的SH-SY5Y细胞损伤的影响及作用机制  

Effect of Sufentanil on the MPP^+-induced SH-SY5Y Cell Damage and Its Mechanism

作　　者：熊昕丽[1] 陈迅 Xiong Xinli;Chen Xun(Department of Neurology, Oriental Hospital Affiliated to Tongji University, Shanghai 200120, China;Anesthesiology Department, Shanghai Changning Maternal and Child Health Hospital, Shanghai 200051, China)

机构地区：[1]同济大学附属东方医院神经内科,上海200120 [2]上海长宁区妇幼保健院麻醉科,上海200051

出　　处：《成都医学院学报》2020年第3期320-327,共8页Journal of Chengdu Medical College

摘　　要：目的探讨舒芬太尼(SF)对1-甲基-4-苯基-吡啶离子(MPP+)诱导的人神经母细胞瘤SH-SY5Y细胞损伤的影响及其可能的分子机制。方法采用流式细胞术检测MPP+对SH-SY5Y细胞凋亡的促进作用以及SF的抑制作用;采用比色法检测丙二醛(MDA)与谷胱甘肽(GSH)的含量;采用实时荧光定量聚合酶链反应(qRT-PCR)与蛋白质印迹技术(Western blot)检测微小RNA-17-5p(miR-17-5p)、甲基化CpG结合蛋白(MECP2)的表达水平;检测抑制miR-17-5p表达或MECP2过表达后细胞中MDA、GSH水平及细胞凋亡率的变化;双荧光素酶报告基因检测miR-17-5p与MECP2的靶向作用关系;Western blot检测B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关蛋白(Bax)的表达水平。结果MPP+可明显促进SH-SY5Y细胞凋亡(P<0.05),Bax蛋白表达水平与MDA水平明显升高(P<0.05),GSH水平与Bcl-2蛋白表达水平明显减低(P<0.05);与MPP+组比较,不同浓度的SF处理后细胞凋亡明显减少(P<0.05),MECP2、Bcl-2表达水平与GSH水平明显升高(P<0.05),miR-17-5p、Bax表达水平与MDA水平明显降低(P<0.05),抑制miR-17-5p表达或MECP2过表达具有相似的作用;双荧光素酶报告实验证实,miR-17-5p可靶向调控MECP2的表达与活性;miR-17-5p过表达可逆转SF对MPP+诱导的SH-SY5Y细胞损伤的保护作用。结论SF可通过调控miR-17-5p/MECP2表达,抑制细胞凋亡及增强其抗氧化能力,从而减轻MPP+诱导的SH-SY5Y细胞损伤。Objective To investigate the effect of sufentanil(SF)on the 1-methyl-4-phenyl-pyridinium(MPP+)-induced damage of human neuroblastoma SH-SY5Y cells and its possible molecular mechanism.Methods The promotion effect of MPP+and the inhibitory effect of SF on SH-SY5Y cell apoptosis was detected by flow cytometry and the content of malondialdehyde(MDA)and glutathione(GSH)was measured by colorimetry.The expression levels of microRNA-17-5p(miR-17-5p)and methyl-CpG-binding protein 2(MECP2)were detected by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR)and Western blot.The changes of the MDA and GSH levels and the apoptosis rate in cells were detected after the inhibition of miR-17-5p expression or MECP2 overexpression.The dual luciferase reporter gene was used to detected the targeting relationship between miR-17-5p and MECP2.The protein expression levels of B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)were detected by Western blot.Results MPP+significantly promoted the apoptosis of SH-SY5Y cells(P<0.05),the Bax protein expression and the MDA level were significantly increased(P<0.05),and the GSH levels and Bcl-2 protein expression levels were significantly decreased(P<0.05).Compared with the MPP+group,the apoptosis was significantly decreased after the treatment with different concentrations of SF(P<0.05),the MECP2 and Bcl-2 expression and the GSH level were significantly increased(P<0.05),the miR-17-5p and Bax expression levels and the MDA level were significantly decreased(P<0.05),and the inhibitation of miR-17-5p expression and MECP overexpression have similar effect.Dual luciferase reporter experiments confirmed that miR-17-5p targeted the regulation of the MECP2 expression and activity.The overexpression of miR-17-5p reversed the protective effect of SF on the MPP+-induced SH-SY5Y cell damage.Conclusion Sufentanil can attenuate the MPP+-induced SH-SY5Y cell damage by regulating the miR-17-5p/MECP2 expression to inhibit the apoptosis and enhance the antioxidant capacity.

关 键 词：舒芬太尼 miR-17-5p MECP2 MPP^+ SH-SY5Y细胞 

分 类 号：R965[医药卫生—药理学]