| 李娟,蔡萃,聂晶,吴宇婧,张东阳.TLR-7对阿尔兹海默细胞模型β淀粉样蛋白表达及炎症反应的调控作用及机制李娟? 蔡萃 聂晶 吴宇婧 张东阳[J].浙江中西医结合杂志,2024,34(4): |
| TLR-7对阿尔兹海默细胞模型β淀粉样蛋白表达及炎症反应的调控作用及机制李娟? 蔡萃 聂晶 吴宇婧 张东阳 |
| The regulation and mechanism of TLR-7 on the expression of amyloid β and inflammatory response in Alzheimer"s cell model |
| 投稿时间：2023-07-18 修订日期：2023-09-20 |
| DOI： |
| 中文关键词: TLR-7 miR-15b 阿尔兹海默症 β淀粉样蛋白 炎症反应 |
| 英文关键词:TLR-7 miR-15b Alzheimer’s disease amyloid β inflammation |
| 基金项目:浙江省医药卫生科研项目 |
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| 中文摘要: |
| [目的]? 探究TLR-7在阿尔兹海默症SH-SY5Y细胞模型中对β淀粉样蛋白表达及神经炎症反应的调控作用及机制。[方法] 构建稳定表达APPswe的阿尔兹海默症模型SH-SY5Y/APPswe细胞,将细胞分为SH-SY5Y组、SH-SY5Y/APPswe组、SH-SY5Y/APPswe+shNC组、SH-SY5Y/APPswe+shTLR7组、SH-SY5Y/APPswe+sh-TLR7+miR-15binhibitor组和SH-SY5Y/APPswe+shTLR7+ inhibitor NC组。用蛋白质免疫印迹检测APP蛋白的表达,用实时荧光定量PCR实验检测TLR-7、miR-15b、BACE1、IL-1β、IL-6和TNF-α mRNA的表达,用酶联免疫吸附实验检测β淀粉样蛋白以及IL-1β、IL-6和TNF-α的分泌；用荧光素酶报告基因实验检测NF-κB活性。[结果] 与SH-SY5Y组相比,SH-SY5Y/APPswe细胞APP表达水平上调,β淀粉样蛋白分泌水平增加(P<0.001)；与SH-SY5Y/APPswe+shNC组相比,SH-SY5Y/APPswe+shTLR7组miR-15b表达水平上调(P<0.001),BACE1 mRNA表达水平下降(P<0.01),β淀粉样蛋白分泌水平下降,IL-1β、IL-6和TNF-α mRNA表达及蛋白分泌水平下降(均P<0.05),NF-κB活性下降(P<0.01)。与SH-SY5Y/APPswe+shTLR7+ inhibitor NC组相比,SH-SY5Y/APPswe+sh-TLR7+miR-15binhibitor组miR-15b表达水平下降(P<0.001),BACE1 mRNA表达水平升高(P<0.05),β淀粉样蛋白分泌水平升高(P<0.01),IL-1β、IL-6和TNF-α mRNA表达及蛋白分泌水平升高(均P<0.05),NF-κB活性升高(P<0.01)。[结论] TLR-7通过miR-15b调控NF-κB信号通路,并影响阿尔兹海默症模型细胞β淀粉样蛋白的分泌及神经炎症反应。guan |
| 英文摘要: |
| Objective?? To investigate the regulatory effect and mechanism of TLR-7 on amyloid β expression and neuroinflammatory response in SH-SY5Y cell model of Alzheimer"s disease. Methods? Constructing SH-SY5Y/APPswe cells, a model of Alzheimer"s disease with stable expression of APPswe, The cells were divided into SH-SY5Y group, SH-SY5Y/APPswe group, SH-SY5Y/APPswe+shNC group, SH-SY5Y/APPswe+shTLR7 group, and SH-SY5Y/APPswe+sh-TLR7+miR-15b inhibitor group and SH-SY5Y/APPswe+shTLR7+ inhibitor NC group. The expression of APP protein was detected by Western blot, the mRNA expressions of TLR-7, miR-15b, BACE1, IL-1β, IL-6 and TNF-α were detected by real-time fluorescent quantitative PCR, and the secretion of β-amyloid and IL-1β, IL-6 and TNF-α were detected by enzyme-linked immunosorbent assay. NF-κB activity was detected by luciferase reporter gene assay. Results? Compared with SH-SY5Y group, the expression level of APP in SH-SY5Y/APPswe cells was up-regulated, and the secretion level of amyloid beta was increased (P<0.001). Compared with SH-SY5Y/APPswe+shNC group, the expression level of miR-15b in SH-SY5Y/APPswe+shTLR7 group was up-regulated (P<0.001), the expression level of BACE1 mRNA was decreased (P<0.01), and the secretion level of amyloid beta was decreased. The mRNA expression and protein secretion levels of IL-1β, IL-6 and TNF-α were decreased (all P<0.05), and the NF-κB activity was decreased (P<0.01). Compared with SH-SY5Y/APPswe+shTLR7+ inhibitor NC group, the expression level of miR-15b was decreased in SH-SY5Y/APPswe+ SHTLR7 +miR-15b inhibitor group (P<0.001). BACE1 mRNA expression level increased (P<0.05), β-amyloid secretion level increased (P<0.01), IL-1β, IL-6 and TNF-α mRNA expression and protein secretion level increased (all P<0.05), NF-κB activity increased (P<0.01). Conclusion? TLR-7 regulates the NF-κB signaling pathway through miR-15b, and influences the secretion of amyloid beta and neuroinflammatory response in Alzheimer"s disease model cells. |
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