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Purpurogallin Reverses Neuronal Apoptosis and Enhances “M2” Polarization of Microglia Under Ischemia via Mediating the miR-124-3p/TRAF6/NF-κB Axis

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Abstract

Purpurogallin (PPG) has been demonstrated to exert an anti-inflammatory function in neurological diseases. This study aimed at investigating the role of PPG on microglial polarization post ischemic stroke as well as the underlying mechanism. Mouse hippocampal neurons HT-22 and microglial BV2 cells were treated by oxygen and glucose deprivation to simulate an in-vitro ischemia model. qRT-PCR and ELISA examined expression of cytokines in microglia. CCK8 and flow cytometry measured HT-22 cell viability and apoptosis, respectively. The levels of miR-124-3p and TRAF6/NF-κB were determined. A mouse cerebral ischemia model was set up using middle cerebral artery occlusion (MCAO) method. After being dealt with PPG, the neurological functions, brain edema, neuronal apoptosis, and microglia activation of the mice were evaluated. As suggested by the results, PPG transformed “M1” to “M2” polarization of BV2 cells, and abated HT-22 cell apoptosis. PPG enhanced the neurological functions, alleviated brain edema, and decreased neuroinflammatory responses, and neuronal apoptosis in the brain lesions of MCAO mice. Furthermore, PPG enhanced miR-124-3p and repressed the TRAF6/NF-κB pathway. miR-124-3p suppressed the TRAF6/NF-κB pathway by targeting TRAF6. Collectively, PPG alleviates ischemia-induced neuronal damage and microglial inflammation by modulating the miR-124-3p/TRAF6/NF-κB pathway.

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Data Availability

The data sets used and analyzed during the current study are available from the corresponding author on reasonable request.

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  1. Department of Neurology, Nanchang First Hospital, No.128 Xiangshan North Road, Donghu District, Nanchang, 330006, Jiangxi, China

    Zongxin Cheng, Xinming Li, Rong Yu & Youqing Deng

  2. Department of Oncology, Jiangxi Maternal and Child Health Hospital, Nanchang, 330006, Jiangxi, China

    Xiaohua Ye

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Conceptualization & Methodology: ZC; Formal analysis and investigation: ZC, XL, XY; Data curation & Resources: RY, YD; Writing—review and editing: ZC. All authors read and approved the final manuscript.

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11064_2022_3752_MOESM1_ESM.tif

Supplementary file1 Supplementary figure 1: PPG repressed neuronal apoptosis induced by microgliaA: The conditioned medium. B: CCK8 assay measured neuronal apoptosis. C: Western blot confirmed the protein profiles of pro-apoptotic proteins (Bax, Bad, and C-caspase3) in HT22 neurons. (TIF 1138 kb)

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Cheng, Z., Li, X., Ye, X. et al. Purpurogallin Reverses Neuronal Apoptosis and Enhances “M2” Polarization of Microglia Under Ischemia via Mediating the miR-124-3p/TRAF6/NF-κB Axis. Neurochem Res 48, 375–392 (2023). https://doi.org/10.1007/s11064-022-03752-4

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