Pyruvate kinase isoform M2 impairs cognition in systemic lupus erythematosus by promoting microglial synaptic pruning via the β-catenin signaling pathway.
Pyruvate kinase isoform M2 impairs cognition in systemic lupus erythematosus by promoting microglial synaptic pruning via the β-catenin signaling pathway.
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丙酮酸激酶亚型 M2 通过 β-连环蛋白信号通路促进小胶质细胞突触修剪，损害系统性红斑狼疮的认知
DOI：
10.1186/s12974-021-02279-9
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发表时间：
2021-10-13
影响因子：
9.3
通讯作者：
Dou H
中科院分区：
文献类型：
--
作者：
Lu L;Wang H;Liu X;Tan L;Qiao X;Ni J;Sun Y;Liang J;Hou Y;Dou H
神经精神系统红斑狼疮（NPSLE）是一种严重的并发症，涉及大脑和认知功能的病理损害。然而，其确切的作用机制仍不清楚。在本研究中，我们探讨了小胶质细胞在NPSLE小鼠认知功能障碍中的作用。我们还分析和比较了狼疮模型小鼠和对照组小鼠海马组织中的代谢物。采用MRL/MpJ-Faslpr （MRL/lpr）雌性小鼠作为NPSLE小鼠模型。代谢组学用于评估海马糖酵解水平。ELISA法检测海马组织中葡萄糖、乳酸、IL-6、IL-1β的含量。基于糖酵解途径，我们发现海马中丙酮酸激酶异构体M2 （PKM2）明显增加。因此，通过qRT-PCR和Western blotting检测PKM2的表达，并通过免疫荧光染色评估PKM2在小胶质细胞（IBA-1+）或神经元（NeuN+）中的定位。流式细胞术检测小胶质细胞的数量和表型；在过表达PKM2的BV2细胞中检测小胶质细胞吞噬和β-连环蛋白信号通路的变化。在体内实验中，用AAV9-shPKM2处理MRL/lpr小鼠。2个月后，采用Morris水迷宫和条件恐惧实验考察小鼠的认知能力；H&E和免疫荧光染色评价脑损伤；流式细胞术检测小胶质细胞表型和功能；采用qRT-PCR、Western blotting和免疫荧光染色监测神经元突触损伤。MRL/lpr狼疮小鼠海马糖酵解升高，同时葡萄糖消耗和乳酸生成增加。此外，我们还观察到狼疮小鼠海马小胶质细胞中PKM2的激活。细胞实验表明，PKM2通过β-catenin信号通路促进小胶质细胞激活和过度激活小胶质细胞吞噬。在体内，通过阻断β-catenin信号通路，aav9 - shpkm2处理的小鼠显示小胶质细胞激活减少，神经元突触丢失减少。此外，小胶质细胞PKM2抑制后，MRL/lpr小鼠的认知功能障碍和脑损伤明显减轻。这些数据表明，小胶质细胞PKM2有可能成为治疗狼疮脑病的新靶点。在线版本包含补充材料，可在10.1186/s12974-021-02279-9获得。
Neuropsychiatric systemic lupus erythematosus (NPSLE) is a severe complication, which involves pathological damage to the brain and cognitive function. However, its exact mechanism of action still remains unclear. In this study, we explored the role of microglia in the cognitive dysfunction of NPSLE mice. We also analyzed and compared the metabolites in the hippocampal tissues of the lupus model and control mice. MRL/MpJ-Faslpr (MRL/lpr) female mice were used as the NPSLE mouse model. Metabolomics was used to assess hippocampal glycolysis levels. Glucose, lactic acid, IL-6, and IL-1β of the hippocampus were detected by ELISA. Based on the glycolysis pathway, we found that pyruvate kinase isoform M2 (PKM2) in the hippocampus was significantly increased. Thus, the expression of PKM2 was detected by qRT-PCR and Western blotting, and the localization of PKM2 in microglia (IBA-1+) or neurons (NeuN+) was assessed by immunofluorescence staining. Flow cytometry was used to detect the number and phenotype of microglia; the changes in microglial phagocytosis and the β-catenin signaling pathway were detected in BV2 cells overexpressing PKM2. For in vivo experiments, MRL/lpr mice were treated with AAV9-shPKM2. After 2 months, Morris water maze and conditional fear tests were applied to investigate the cognitive ability of mice; H&E and immunofluorescence staining were used to evaluate brain damage; flow cytometry was used to detect the phenotype and function of microglia; neuronal synapse damage was monitored by qRT-PCR, Western blotting, and immunofluorescence staining. Glycolysis was elevated in the hippocampus of MRL/lpr lupus mice, accompanied by increased glucose consumption and lactate production. Furthermore, the activation of PKM2 in hippocampal microglia was observed in lupus mice. Cell experiments showed that PKM2 facilitated microglial activation and over-activated microglial phagocytosis via the β-catenin signaling pathway. In vivo, AAV9-shPKM2-treated mice showed decreased microglial activation and reduced neuronal synapses loss by blocking the β-catenin signaling pathway. Furthermore, the cognitive impairment and brain damage of MRL/lpr mice were significantly relieved after microglial PKM2 inhibition. These data indicate that microglial PKM2 have potential to become a novel therapeutic target for treating lupus encephalopathy. The online version contains supplementary material available at 10.1186/s12974-021-02279-9.
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DOI：
10.1016/j.omtm.2021.01.006
发表时间：
2021-03-12
期刊：
Molecular therapy. Methods & clinical development
影响因子：
--
作者：
Eykens C;Rossaert E;Duqué S;Rué L;Bento-Abreu A;Hersmus N;Lenaerts A;Kerstens A;Corthout N;Munck S;Van Damme P;Holt MG;von Jonquires G;Klugmann M;Van Den Bosch L;Robberecht W
通讯作者：
Robberecht W
影响因子：
10.6
作者：
Crupi, Rosalia;Cambiaghi, Marco;Battaglia, Fortunato
通讯作者：
Battaglia, Fortunato
影响因子：
64.8
作者：
Bialas, Allison R.;Presumey, Jessy;Carroll, Michael C.
通讯作者：
Carroll, Michael C.
小胶质细胞在发育和晚年中枢神经系统病理学中发挥着关键作用。
影响因子：
12.7
作者：
Derecki NC;Katzmarski N;Kipnis J;Meyer-Luehmann M
通讯作者：
Meyer-Luehmann M
影响因子：
--
作者：
Brooks, William M.;Sibbitt, Wilmer L., Jr.;Kornfeld, Mario;Jung, Rex E.;Bankhurst, Arthur D.;Roldan, Carlos A.
通讯作者：
Roldan, Carlos A.