| 赵 琳,于丰祥,谭小芳,宫 平,胡竹琴,邱 瑜,李 娟.SK3钾通道通过PI3K/Akt-ERK通路介导Cu2+-Aβ复合物所致小胶质细胞激活[J].,2023,(13):2401-2406 |
| SK3钾通道通过PI3K/Akt-ERK通路介导Cu2+-Aβ复合物所致小胶质细胞激活 |
| SK3 Potassium Channels Mediate Microglial Activation Caused by Cu2+-Aβ Complex via the PI3K/Akt-ERK Signaling Pathway |
| 投稿时间：2023-03-29 修订日期：2023-04-25 |
| DOI：10.13241/j.cnki.pmb.2023.13.001 |
| 中文关键词: SK3通道 Cu-Aβ复合物 PI3K/Akt-ERK信号通路 小胶质细胞 |
| 英文关键词: SK3 channels Cu-Aβ complex PI3K/Akt-ERK signaling pathway Microglia |
| 基金项目:国家自然科学基金项目（82073854） |
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| 中文摘要: |
| 摘要 目的：探讨Ca2+激活的小电导SK3钾通道在Cu2+-Aβ复合物（Cu-Aβ）所致小胶质细胞激活中的作用及下游信号通路。方法：应用Cu-Aβ激活BV2小胶质细胞，采用ELISA和Amplex Red试剂盒检测细胞培养上清中肿瘤坏死因子（TNF-α）和过氧化氢（H2O2）的含量，应用qPCR和Western blot检测钾通道mRNA和蛋白水平及相关信号通路蛋白的磷酸化。结果：（1）应用不同离子通道阻断剂以及不同亚型钾通道阻断剂预处理的实验结果表明，SK3通道可能介导了Cu-Aβ所致的小胶质细胞激活。（2）qPCR和Western blot检测结果表明，Cu-Aβ可上调小胶质细胞内SK3 mRNA和蛋白表达。（3）通过转染SK3-siRNA下调小胶质细胞内SK3表达水平，结果表明，下调SK3表达后显著抑制Cu-Aβ所致的小胶质细胞激活。（4）应用特异性信号分子阻断剂预处理的实验结果表明，PI3K/Akt信号和 ERK信号均参与了Cu-Aβ所致的小胶质细胞激活。（5）应用相关信号分子阻断剂预处理的实验结果进一步表明，在介导Cu-Aβ诱发的小胶质细胞激活过程中，SK3通道位于PI3K/Akt-ERK信号通路的上游。结论：SK3通道通过其下游的PI3K/Akt-ERK信号通路介导Cu-Aβ所致的小胶质细胞炎症反应。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the role of Ca2+-activated small conductance SK3 potassium channels in Cu-Aβ complex-induced microglial activation and the downstream signaling pathway. Methods: The Cu-Aβ complex-induced release of tumor necrosis factor-α(TNF-α) and hydrogen peroxide (H2O2) in cell culture supernatants were detected using ELISA and the Amplex Red reagent. The levels of mRNA and protein of potassium channels as well as phosphorylation levels of related signaling molecules were detected by qPCR and Western blot. Results: (1) The experimental results of pretreatment with different ion channel blockers or potassium channel blockers of different subtypes showed that SK3 channels may mediate microglial activation caused by Cu-Aβ complex. (2) The results of qPCR and Western blot detection showed that Cu-Aβ complex could upregulate SK3 mRNA and protein expression in microglia. (3) The application of SK3-siRNA to knock down SK3 expression showed that the downregulation of SK3 expression significantly inhibited microglial activation caused by Cu-Aβ complex. (4) The experimental results of Western blot detection showed that PI3K/Akt signaling and ERK signaling were involved in microglial activation caused by Cu-Aβ complex. (5) The results of experiments using specific blockers of signaling molecules further showed that SK3 channels may lie upstream of the PI3K/Akt-ERK signaling pathway during the microglial activation induced by the Cu-Aβ complex. Conclusion: SK3 channels mediate microglial inflammatory responses induced by Cu-Aβ complex through the downstream PI3K/Akt-ERK signaling pathway. |
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