6th Edition of Neurology World Conference 2026

Abstract

Background: Ginkgolide B (GB) has been shown to have a neuroprotective effect on ischemic stroke,  neurons against ischemia-induced apoptosis, white matter lesion caused by chronic cerebral hypoperfusion or NLRP3 inflammasome in the activated BV2 cells induced by Aβ1-42. However, it still remains unknown if Ginkgolide B can modulate the oxidative stress in the microglial activation induced by Lipopolysaccharide (LPS).

Objective: To know the effect of GB on microglial activation, like inducible nitric oxidative synthase (iNOS) and nitric oxide (NO)upregulation by LPS.

Method: The primary microglia were obtained from the primary mixed-glial cultures, which were derived from 2-3d SD rats, after 10-day growth using the shake-off method. The primary microglia were seeded into the 24-well plates at the density of 3.5 x10⁵ cells/well while BV2 were seeded at the density of 1x 10⁵ cells /well, and then they were incubated at 5% CO₂, 37℃ for 24h before any addition. Based on the experience in our previous exploration of GB on oxidative stress in the mixed glia, 10μM of Ginkgolide B was used to the activated microglia induced by 100ng/ml of LPS in this study. Additionally, SB inhibitors related to MAPK signaling pathways as exploring working mechanisms or Aspirin as a positive control were enrolled into in BV2 experiments. The cells or supernatant were harvested after co-incubation for 12h, 24h, or 36h. The production of nitric oxide in the supernatant released via microglia was detected by Griess assay, and iNOS expression in the cells was measured by immunofluorescent antibody.

Result: The results showed that both primary microglia or BV2 were activated by 100ng/ml of LPS at 12h, 24h, or 36h, which included the upregulation of NO and iNOS in a time-dependent manner. When GB was added into the cells for 12h, 24h, or 36h, NO and iNOS were down-regulated, which showed a time-dependent manner. Compared to LPS, NO value down-regulation by GB was significant (P＜0.05 at 24h- or 36h-treatment of GB). Interestingly, we measured some inflammatory molecules such as COX-2 or TNF-α in cell lysates at the same time points by Western Blotting analysis and found that GB could down-regulate the inflammatory response induced by LPS as well.

Conclusion: Ginkgolide B could down-regulate the oxidative stress in not only the primary mixed glia but also in the primary rat microglia or mouse microglial cell line BV2 induced by LPS, which suggested that GB potentially has pharmacological value in intervention to CNS disorders with oxidative stress and inflammation. This study provided more evidence to use GB in the therapy at the early stage of neurodegenerative diseases, including Alzheimer’s disease or PD.