Panax Notoginseng Saponins-induced Autophagy-mediated Microglial Polarization via Activation of the AMPK Signaling Pathway and Its Effects on Neurons in A Cellular Model of Neuropathic Pain

Ying TAN; Wendong CHEN; Lanqing PU; Kun YANG; Ziting REN; Jun WANG; Haixia JIANG

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Ying TAN, Wendong CHEN, Lanqing PU, Kun YANG, Ziting REN, Jun WANG, Haixia JIANG. Panax Notoginseng Saponins-induced Autophagy-mediated Microglial Polarization via Activation of the AMPK Signaling Pathway and Its Effects on Neurons in A Cellular Model of Neuropathic Pain[J]. Journal of Kunming Medical University.

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Panax Notoginseng Saponins-induced Autophagy-mediated Microglial Polarization via Activation of the AMPK Signaling Pathway and Its Effects on Neurons in A Cellular Model of Neuropathic Pain

Department of Anesthesiology，The First Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650032，China

Received Date: 2026-06-17
Available Online: 2026-09-30

Abstract

Abstract

Objective To investigate the effects of Panax notoginseng saponins （PNS） on microglial polarization and its effects on neuronal cells in a neuropathic pain （NP） cell model through autophagy regulation mediated by the AMPK signaling pathway. Methods BV2 cells were pretreated with PNS at different concentrations （0, 50, 100, 200, and 400 μg/mL）, the AMPK activator metformin, or the AMPK inhibitor dorsomorphin, followed by co-stimulation with LPS and IFN-γ for 24 h to establish an NP cell model. Flow cytometry, JC-1 staining, DCFH-DA fluorescent probe, Western blot, and ELISA were employed to assess the effects of PNS on BV2 cell polarization and its modulation of the AMPK/mTOR signaling pathway. Mouse hippocampal neurons （MHN） were co-cultured with BV2 cells subjected to the above treatments. The effects of BV2 cell polarization on MHN cell viability and apoptosis were evaluated using the CCK-8 assay and the Annexin V-FITC/PI apoptosis detection kit. Results LPS and IFN-γ treatment increased the proportion of INOS-positive BV2 cells （P < 0.001）, increased TNF-α and IL-1β levels （P < 0.001）, decreased the proportion of CD206-positive cells （P < 0.001） and the concentrations of IL-10 and IL-4 （P < 0.001）, reduced mitochondrial membrane potential （P < 0.001）, increased ROS levels and P62 protein expression （P < 0.001）, and inhibited LC3Ⅱ/Ⅰ expression （P < 0.001）. Meanwhile, this treatment inhibited p-AMPK expression （P<0.001）, increased p-mTOR expression （P < 0.001）, decreased the viability of cocultured MHNs （P < 0.001）, and increased their apoptosis rate （P < 0.001）. Pretreatment with 3-MA inhibited LC3 II/I expression （P < 0.05） and increased P62 protein expression （P < 0.01）. Pretreatment with PNS or metformin promoted p-AMPK expression （P < 0.01）, inhibited p-mTOR expression （P < 0.001）, decreased the proportion of iNOS-positive BV2 cells and the levels of TNF-α and IL-1β induced by LPS and IFN-γ （P < 0.001）, and increased the proportion of CD206-positive cells and the concentrations of IL-10 and IL-4 （P < 0.001）. It also increased mitochondrial membrane potential and autophagy in BV2 cells （P < 0.001）, reduced ROS levels （P < 0.01）, increased cocultured MHN viability, and reduced apoptosis （P < 0.05）. Pretreatment with dorsomorphin further enhanced the effects of LPS and IFN-γ on increasing the proportion of iNOS-positive BV2 cells and the levels of TNF-α and IL-1β （P < 0.05）, while reducing the proportion of CD206-positive cells and the levels of IL-10 and IL-4 （P < 0.05）. It also decreased mitochondrial membrane potential, promoted ROS generation （P < 0.01）, reduced cocultured MHN viability, and aggravated apoptosis （P < 0.05）. Combined pretreatment with PNS and metformin enhanced the protective effects of metformin on BV2 and MHN cells （P < 0.001）, whereas combined pretreatment with PNS and dorsomorphin partially reversed the deleterious effects of dorsomorphin （P < 0.05）. Conclusion PNS inhibits LPS/IFN-γ-induced M1 polarization of BV2 cells, alleviates mitochondrial injury, enhances MHN cell viability, and reduces their apoptosis by activating AMPK and inhibiting the mTOR signaling pathway.
- Microglia,
- Autophagy,
- Panax notoginseng saponins,
- AMPK signaling pathway,
- Neurons

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References(31)

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