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A Cell Culture Model for Studying the Role of Neuron-Glia Interactions in Ischemia
Published on: November 14, 2020
Enteric glial cells exert neuroprotection from hyperglycemia-induced damage via Akt/GSK3β pathway
1Department of Pharmacy, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan.
Objective:
Enteric glial cells (EGCs) can activate multiple pathways to inhibit the deleterious effects of acute and chronic insults. Our aim was to test the effect of EGCs on hyperglycemia-induced neuron damage and its underlying intracellular mechanisms.
Methods:
A coculture model composed of EGCs and neuroblastoma cells (SH-SY5Y) was established to examine glial-mediated neuroprotection under high glucose conditions. The cell counting assay kit CCK-8 was used to measure cell viability. Flow cytometry was used to measure the induction of reactive oxygen species (ROS), change of mitochondrial membrane potential (MMP), cell cycle distribution, and apoptosis. The expressions of cyclin D1, cyclin E2, Bax, cleaved caspase-3, AKT, p-AKT, GSK-3β, and p-GSK-3β were tested using western blot.
Results:
Exposure to high glucose (≥35 mM) reduced the viability of SH-SY5Y cells in a concentration- and time-dependent manner. Meanwhile, enhanced ROS generation and decrease of MMP were observed in SH-SY5Y cells when treated with high glucose. Furthermore, high glucose also caused SH-SY5Y cells arrest in G2 phase and apoptosis, accompanied by decreasing cyclin D1 and E2, and upregulating Bax and cleaved caspase-3. Coculture EGC lines or EGC-conditioned medium with SH-SY5Y prevented the neurotoxic effects. The p-AKT/AKT and p-GSK-3β/GSK-3β ratios were dramatically decreased in SH-SY5Y cells after high glucose incubation, which was restored after coculture with EGCs.
Conclusions:
EGCs can protect neurons from hyperglycemia-induced injury by activating the Akt/GSK-3β pathway.
Insights
Enteric glial cells (EGCs) protect neurons from high glucose damage by activating the Akt/GSK-3β pathway. This study reveals a novel mechanism for glial-mediated neuroprotection against hyperglycemia-induced injury.
Area of Science:
- Neuroscience
- Cell Biology
- Glial Cell Biology
Background:
- Enteric glial cells (EGCs) possess neuroprotective capabilities against various insults.
- Hyperglycemia is a known risk factor for neuronal damage, particularly in the enteric nervous system.
- Understanding the mechanisms of EGC-mediated neuroprotection is crucial for developing therapeutic strategies.
Purpose of the Study:
- To investigate the protective effects of EGCs against hyperglycemia-induced neuronal damage.
- To elucidate the intracellular signaling pathways involved in EGC-mediated neuroprotection.
Main Methods:
- A co-culture model of EGCs and SH-SY5Y neuroblastoma cells was established.
- Cell viability, reactive oxygen species (ROS) generation, mitochondrial membrane potential (MMP), cell cycle, and apoptosis were assessed.
- Western blotting was used to analyze the expression of key proteins in the Akt/GSK-3β pathway.
Main Results:
- High glucose significantly reduced SH-SY5Y cell viability, increased ROS, decreased MMP, and induced apoptosis and G2 cell cycle arrest.
- EGC co-culture or conditioned medium protected SH-SY5Y cells from high glucose-induced neurotoxicity.
- High glucose decreased p-AKT/AKT and p-GSK-3β/GSK-3β ratios, which were restored by EGCs.
Conclusions:
- EGCs effectively protect neurons against hyperglycemia-induced injury.
- The Akt/GSK-3β signaling pathway is a key mediator of EGC-mediated neuroprotection.
- EGCs represent a potential therapeutic target for managing hyperglycemia-related neurological complications.
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