摘要： 目的探讨跨膜蛋白16F （TMEM16F）对β淀粉样蛋白（Aβ25-35）诱导的阿尔茨海默病（AD）细胞模型铁死亡的影响。方法将SH-SY5Y细胞分为对照组、模型组、siRNA阴性对照组（siRNA-nc组）、siRNA-TMEM16F转染组（siRNA-TMEM16F组）。采用CCK-8检测Aβ25-35对SH-SY5Y细胞存活率的影响，Western blotting检测各组TMEM16F、AD相关指标（Aβ、p-Tau）、铁死亡相关蛋白（ACSL4、GPX4、Fpn1）的表达情况，免疫荧光法检测各组ACSL4表达情况，荧光探针法检测各组活性氧（ROS）水平。结果 Aβ25-35≥20 μmol/L作用≥12 h时对细胞有显著损伤作用。与对照组相比，模型组中TMEM16F、Aβ、p-Tau、ACSL4以及ROS水平显著增高，而GPX4、Fpn1水平显著降低（P<0.05）；沉默TMEM16F后，Aβ、p-Tau、ACSL4及ROS表达受到抑制，而GPX4、Fpn1表达水平上升（P<0.05）。结论沉默TMEM16F能够抑制Aβ25-35诱导的SH-SY5Y细胞发生铁死亡。

跨膜蛋白16F对阿尔茨海默病细胞模型铁死亡的调控作用

胡晓莹¹, 张跃骜², 崔志强¹, 顾颖¹, 杨拓¹, 张文强¹, 张慧予³

1. 中国医科大学 附属第四医院神经内科, 沈阳 110032;
2. 中国医科大学 临床一系2020级82班, 沈阳 110122;
3. 中国医科大学 附属第四医院老年病科, 沈阳 110032

收稿日期:2021-12-22出版日期:2022-05-30发布日期:2022-05-28
通讯作者:张慧予
作者简介:胡晓莹(1994-),女,硕士研究生.
基金资助:中国博士后科学基金（2020M681012）；辽宁省重点研发计划（2020JH2/10300044）；沈阳市科学技术计划（20-205-4-050）；大学生创新创业训练计划（202110159020）


关键词: 阿尔茨海默病, 跨膜蛋白16F, 铁死亡
Abstract: Objective To investigate the effect of the transmembrane protein 16F (TMEM16F) on ferroptosis of β-amyloid protein (Aβ25-35) -induced SH-SY5Y cells mimicking Alzheimer disease (AD). Methods SH-SY5Y cells were divided into the control group,model group (Aβ25-35),siRNA-nc group (siRNA-nc+Aβ25-35),and siRNA-TMEM16F group (siRNA-TMEM16F+Aβ25-35). The effect of Aβ25-35 on the viability of SH-SY5Y cells was detected by cell counting kit-8 (CCK-8). The expression of TMEM16F protein,AD- related indicators,and ferroptosis-related indicators were detected by western blotting. The expression of ACSL4 was detected by immunofluore- scence. The reactive oxygen species (ROS) level was detected by fluorescence probe. Results CCK8 results showed that SH-SY5Y cells were significantly injured by 20 μmol/L Aβ25-35 after 12 h. Western blotting and immunofluorescence found that the expression of Aβ and p-Tau was increased and the expression of ACSL4 was upregulated in the model group compared with the control group. The expression of GPX4 and Fpn1 protein was decreased in the model group (P< 0.05). The immunofluorence result of ACSL4 was consistent with those before. ROS level was improved in model group. After TMEM16F silencing,AD- and ferroptosis-related indicators were inhibited compared with the siRNA-nc group (P< 0.05). Conclusion Silencing TMEM16F can inhibit ferroptosis in Aβ25-35-induced SH-SY5Y cells.
Key words: Alzheimer disease, transmembrane protein 16F, ferroptosis
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