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人参二醇对APP-SH-SY5Y细胞中Tau蛋白磷酸化及Fyn/GluN2B信号通路的影响
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| 篇名： | 人参二醇对APP-SH-SY5Y细胞中Tau蛋白磷酸化及Fyn/GluN2B信号通路的影响 |
| TITLE： | Effects of Panaxadiol on Tau Protein Phosphorylation and Fyn/GluN 2B Signaling Pathway in APP-SH-SY 5Y Cells |
| 摘要： | 目的：研究人参二醇（PD）对转染APP基因的人神经母细胞瘤细胞SH-SY5Y（APP-SH-SY5Y）中Tau蛋白磷酸化的影响及其作用机制。方法：采用分子对接技术验证PD与非受体酪氨酸激酶Fyn的靶向性。在体外培养SH-SY5Y细胞，构建APP-SH-SY5Y细胞模型和绿色荧光蛋白（GFP）-SH-SY5Y细胞模型（对照细胞），通过检测细胞中β淀粉样蛋白（Aβ1-42）的表达来验证APP-SH-SY5Y细胞模型是否构建成功。以GFP-SH-SY5Y细胞为对照，采用CCK-8法检测5、10、20、30、40μmol/L的PD和125、250、500、1000、2000nmol/L的PP2（Fyn抑制剂，阳性对照）作用24h对APP-SH-SY5Y细胞存活率的影响，以确定最佳给药浓度。以APP-SH-SY5Y细胞为对象，分别检测最佳浓度PD、PP2作用24后细胞中Ca2+浓度及磷酸化Tau蛋白（p-Tau）/Tau、磷酸化非受体酪氨酸激酶Src（p-Src）/Fyn、磷酸化谷氨酸受体2B（p-GluN2B）/GluN2B比值。结果：分子模拟对接结果显示，PD可靶向结合Fyn蛋白。与GFP-SH-SY5Y细胞比较，APP-SH-SY5Y细胞中Aβ1-42蛋白表达水平显著升高（P＜0.01）。PD和PP2的最佳作用浓度分别为20μmol/L和500nmol/L。20μmol/LPD、500nmol/LPP2均可显著升高细胞的存活率，显著降低细胞中Ca2+浓度以及p-Tau/Tau、p-Src/Fyn、p-GluN2B/GluN2B比值。结论：PD可通过抑制Fyn/GluN2B信号通路来降低APP-SH-SY5Y细胞中Ca2+浓度及Tau蛋白的磷酸化水平。 |
| ABSTRACT： | OBJECTIVE：To study the effects and mechanism of panaxadiol （PD） on Tau protein phosphorylation in the SH-SY5Y cells transfected with APP gene（APP-SH-SY5Y）. METHODS ：The target of PD and non-receptor tyrosine kinases Fyn was verified by molecular docking. SH-SY 5Y cells were cultured in vitro ，and the APP-SH-SY 5Y cell models and green fluorescent （GFP）-SH-SY5Y cell model （control cell ）was constructed. The expression of Aβ1-42 was detected so as to verify the success of APP-SH-SY5Y cell model. Taking GFP-SH-SY 5Y cells as control ，the effects of 5，10，20，30，40 μmol/L PD and 125，250， 500，1 000，2 000 nmol/L PP 2（Fyn inhibitor ，positive control ）on the survival rate of APP-SH-SY 5Y cells were detected by CCK-8 assay after treated for 24 h，so as to confirm the optimal concentration. The concentration of Ca 2 + ，the ratio fophosphorylated Tau protein （p-Tau）/Tau，phosphorylatedn Src（p-Src）/Fyn and phosphorylated glutamate receptor 2B（p-GluN2B）/ GluN2B were detected in APP-SH-SY 5Y cells after trated with the optimal concentration of PD and PP 2 for 24 h. RESULTS ：The results of molecular simulation docking showed that PD could target Fyn protein. Compared with GFP-SH-SY 5Y cells ，the protein expression of Aβ1-42 in APP-SH-SY 5Y cell were increased significantly （P＜0.01）. The optimal concentration of PD and PP 2 were 20 μmol/L and 500 nmol/L. The 20 μmol/L PD and 500 nmol/L PP 2 could increase the survival rate of the cells and reduced the concentration of Ca 2+，the ratio of p-Tau/Tau ，p-Src/Fyn，and p-GluN 2B/GluN2B. CONCLUSIONS：PD can reduce the the phosphorylation of Tau protein through inhibiting Fyn/GluN 2B signaling pathway. |
| 期刊： | 2021年第32卷第12期 |
| 作者： | 梁喜才，蔺莹，肖洪贺，孔亮，杨静娴 |
| AUTHORS： | LIANG Xicai，LIN Ying，XIAO Honghe ，KONG Liang，YANG Jingxian |
| 关键字： | 人参二醇；转染APP基因的人神经母细胞瘤细胞 SH-SY5Y；Tau蛋白；磷酸化；非受体酪氨酸激酶 Fyn/谷氨酸受体2B信 |
| KEYWORDS： | Panaxadiol；APP-SH-SY5Y cells ；Tau protein ；Phosphorylation；Fyn/GluN2B signaling pathway |
| 阅读数： | 250 次 |
| 本月下载数： | 9 次 |
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