山莨菪碱通过lncRNA LINC00657/miR-496通路对MPP+诱导SK-N-SH 细胞损伤的保护作用研究
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Protective Effect of Anisodamine on SK- N- SH Cell Injury Induced by MPP + Through lncRNA LINC00657/miR-496 Pathway
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    摘要:

    目的:探讨山莨菪碱(Ani) 对1-甲基-4-苯基吡啶离子(MPP+) 诱导SK-N-SH 细胞损伤的作用及分子机制。方法:将SK-N-SH 细胞分为Con 组,MPP+组,MPP+Ani-L、M、H 组,MPP++si-NC 组,MPP++si-LINC00657 组,MPP++Ani+pcDNA 组,MPP++Ani+pcDNA-LINC00657 组。试剂盒检测丙二醛(MDA) 含量和谷胱甘肽(GSH) 活性;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot) 法检测Bcl-2、Bax 蛋白表达;实时荧光定量PCR(RT-qPCR) 检测LINC00657 和miR-496的表达;双荧光素酶报告实验检测LINC00657 和miR-496 的靶向关系。结果:不同浓度山莨菪碱处理后,MPP+诱导SK-N-SH细胞中MDA 含量降低,GSH 活性升高,细胞凋亡率降低,Bcl-2 表达水平升高,Bax 表达水平降低,LINC00657 表达水平降低,miR-496 表达水平升高,且呈浓度依赖性(P<0.05)。LINC00657 靶向调控miR-496 的表达;抑制LINC00657 表达后,MPP+诱导SK-N-SH 细胞中MDA 含量降低,GSH 活性升高,细胞凋亡率降低,Bcl-2 表达水平升高,Bax 表达水平降低(P<0.05)。LINC00657 过表达逆转了山莨菪碱对MPP+诱导SK-N-SH 细胞损伤的作用。结论:山莨菪碱通过调控lncRNA LINC00657/miR-496 通路对MPP+诱导SK-N-SH 细胞损伤起保护作用。

    Abstract:

    Abstract:Objective:To investigate the effect and molecular mechanism of anisodamine(Ani) on MPP+-induced SK-NSH cell damage. Methods:SK-N-SH cells were divided into Con group, MPP+ group,MPP++Ani-L、M、H group, MPP++ si-NC group, MPP++si-LINC00657 group, MPP++Ani+pcDNA group, MPP++Ani+pcDNA-LINC00657 group. Kit to detect malondialdehyde(MDA) content and glutathione(GSH) activity,flow cytometry to detect apoptosis,Western blot method to detect Bcl- 2 and Bax protein expression,real- time fluorescence quantitative PCR(RT- qPCR) detection of LINC00657 and miR- 496 expression, dual luciferase report test to detect the targeting relationship between LINC00657 and miR- 496. Results:After treatment with anisodamine at different concentrations, MDA content was decreased in MPP+-induced SKN- SH cell, GSH activity was increased, apoptosis rate was decreased, Bcl-2 expression was increased, Bax expression was decreased, LINC00657 expression was decreased, expression of miR- 496 was increased, in a concentrationdependent manner(P<0.05). LINC00657 regulates the expression of miR-496, after inhibiting the expression of LINC00657, MDA content was decreased in MPP+-induced SK-N-SH cell, GSH activity was increased, apoptosis rate was decreased, Bcl-2 expression was increased, Bax expression was decreased(P<0.05). LINC00657 overexpression reversed the effect of anisodamine on MPP +- induced SK- N- SH cell damage. Conclusion: Anisodamine protects SK- N- SH cells from MPP +- induced injury by regulating the lncRNA LINC00657/miR-496 pathway.

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齐献忠,邢英瀛,秦慧兵.山莨菪碱通过lncRNA LINC00657/miR-496通路对MPP+诱导SK-N-SH 细胞损伤的保护作用研究[J].新中医,2021,53(21):7-12

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  • 在线发布日期: 2021-11-12
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