PERK-mediated inhibition of endoplasmic reticulum stress in lipopolysaccharide-induced inflammatory responses in microglia

doi:10.16098/j.issn.0529-1356.2025.04.006

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Acta Anatomica Sinica ›› 2025, Vol. 56 ›› Issue (4) : 421-430. DOI: 10.16098/j.issn.0529-1356.2025.04.006

PERK-mediated inhibition of endoplasmic reticulum stress in lipopolysaccharide-induced inflammatory responses in microglia

DING Jia-xin¹ Lü Meng-jun² LU Lin-xia¹ JIFU Ci-li¹ XIA Jun¹ WANG Jing-tao^1*

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Abstract

Objective To explore the role of the protein kinase R-like endoplasmic reticulum kinase (PERK)-mediated endoplasmic reticulum stress pathway in a model of lipopolysaccharide(LPS)-induced microglia inflammation. Methods To investigate its effects on endoplasmic reticulum (ER) stress, an inflammation model of microglia was established by stimulating with LPS at gradient concentrations for 24 hours and with 1 mg/L LPS for different durations. Cell viability was assessed by the CCK-8 assay; The mRNA and protein expression levels of related inflammatory factors were measured by Real-time PCR and ELISA kits. Cellular oxidative stress was evaluated by detecting reactive oxygen species (ROS), and Realtime PCR and Western blotting were used to examine the mRNA and protein expression levels of ER stress pathway markers associated with inflammation. Results 1. The effects of different concentrations of LPS on cell viability and morphology were not statistically significant after acting on BV-2 cells for 24 hours (P>0.05); 2. 1 mg/L LPS incubated with BV-2 cells for different times and the cell viability decreased with the increase of time; 3. Compared with the 0 hour group, the levels of pro-inflammatory cytokine interleukin (IL)-1β, tumor necrosis factor-α (TNF-α) mRNA and protein expression increased significantly (P<0.05) in the LPS-stimulated 9 hours, 12 hours, and 24 hours groups, and the inflammation model was successfully established; 4. Compared with the 0 hour group, the protein and mRNA expression levels of the endoplasmic reticulum stress pathway-related indexes in the LPS-stimulated 9 hours, 12 hours, and 24 hours groups increased significantly (P<0.01), which showed the time-dependence; 5. After adding the PERK inhibitor GSK2606414, the mRNA and protein expression levels of endoplasmic reticulum stress-related indicators in the PERK inhibitor group were significantly reduced compared with those in the LPS group (P<0.05); 6. The mRNA and protein expression levels of pro-inflammatory cytokines and the fluorescence intensity of ROS in the PERK inhibitor group were significantly reduced compared with those in the LPS group (P<0.01). Conclusion Targeting PERK-mediated endoplasmic reticulum stress inhibits LPS-induced inflammatory responses in microglia.

Key words

Inflammation / Endoplasmic reticulum stress / Protein kinase R-like endoplasmic reticulum kinase / Microglia / Real-time PCR / Western blotting

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DING Jia-xin Lü Meng-jun LU Lin-xia JIFU Ci-li XIA Jun WANG Jing-tao. PERK-mediated inhibition of endoplasmic reticulum stress in lipopolysaccharide-induced inflammatory responses in microglia[J]. Acta Anatomica Sinica. 2025, 56(4): 421-430 https://doi.org/10.16098/j.issn.0529-1356.2025.04.006

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