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Updated: Mar 18, 2026
Characterization and Isolation of Mouse Primary Microglia by Density Gradient Centrifugation
Published on: February 16, 2018
Intermittent hypoxia reduces microglia proliferation and induces DNA damage in vitro
Song Liu1, Zhonghua Wang2, Bo Xu3
1Department of Respiratory Medicine, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China, 200092.
Objectives:
Intermittent hypoxia (IH), caused by obstructive sleep apnea (OSA), could cause hippocampus or neuron damage through multiple signaling pathways, while the underlying mechanisms are still unclear. Thus, the present study aimed to explore the effect of IH on the biological functions of microglia cells.
Materials And Methods:
Cell proliferation of BV2 cells after exposure to IH were observed by MTT assay and then DNA damage was detected by comet assay. RNA-sequencing assay was performed in cells under IH condition and normal conditions to find out the differentially expressed genes, which were further confirmed by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot assay.
Results:
As results, IH inhibited the proliferation of BV2 cells, as well as caused DNA damage. RNA-sequencing assay revealed 4 differentially expressed genes (p21, Cyclin D1, Cyclin E2, and Gadd45α) which were associated with the network of P53 signaling pathways in BV2 cells, among which, p21 and Gadd45α were dramatically increased while Cyclin D1 and Cyclin E2 were both decreased significantly. Moreover, inflammatory factors including IL-6, TNF-α and iNOS were significantly up-regulated in microglia cells under IH conditions for 8 hr.
Conclusion:
Our results indicated that IH could inhibit cyclin D1 and cyclin E2 expression via initiating multiple P53 pathways, which further blocked cell cycle transition and attenuated proliferative capability of BV2 cells. Meanwhile, IH activated inflammation reactions in BV2 cells. Present study elaborate the effects of IH on biological functions of microglia and provide theoretical foundation for further study on new therapy methods for OSA.
Insights
Intermittent hypoxia (IH) from obstructive sleep apnea (OSA) impairs microglia cell proliferation and DNA, activating inflammation. IH affects P53 pathways, altering cell cycle genes and providing insights into OSA-related neurological damage.
Area of Science:
- Neuroscience
- Cell Biology
- Sleep Medicine
Background:
- Obstructive sleep apnea (OSA) is linked to neurological damage via intermittent hypoxia (IH).
- The precise mechanisms by which IH affects microglia, key immune cells in the brain, remain largely unknown.
- Understanding microglia's response to IH is crucial for developing therapies for OSA-related cognitive impairment.
Purpose of the Study:
- To investigate the impact of intermittent hypoxia (IH) on the biological functions of microglia cells.
- To elucidate the molecular pathways involved in IH-induced changes in microglia.
- To explore the potential role of P53 signaling in mediating IH effects on microglia.
Main Methods:
- BV2 microglia cells were exposed to IH conditions.
- Cell proliferation was assessed using MTT assay.
- DNA damage was evaluated by comet assay.
- RNA sequencing identified differentially expressed genes, with validation by RT-PCR and Western blot.
- Inflammatory markers were quantified.
Main Results:
- IH significantly inhibited BV2 cell proliferation and induced DNA damage.
- RNA sequencing revealed differential expression of P53 pathway genes: p21 and Gadd45α were upregulated, while Cyclin D1 and Cyclin E2 were downregulated.
- IH exposure led to a significant increase in inflammatory factors (IL-6, TNF-α, iNOS).
Conclusions:
- IH inhibits microglia proliferation by downregulating Cyclin D1 and Cyclin E2 via P53 pathways, impeding cell cycle progression.
- IH activates inflammatory responses in microglia.
- These findings offer a theoretical basis for novel therapeutic strategies targeting microglia in OSA.
