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Activation of BV2 microglia by lipopolysaccharide triggers an inflammatory reaction in PC12 cell apoptosis through a toll-like receptor 4-dependent pathway

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Cell Stress and Chaperones Aims and scope

Abstract

Microglia play an important role in neuronal protection and damage. However, the molecular and cellular relationship between microglia and neurons is unclear. We carried out a prospective study to detect that activation of BV2 microglia induced PC12 cell apoptosis in vitro through the TLR4/adapter protein myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway. BV2 microglia were treated with different concentrations of LPS for 24 h. Western blot was utilized to detect the expression of TLR4 and the downstream signaling pathway. The level of inflammatory mediator was quantified using a specific ELISA kit. The supernatant of 10 μg/ml LPS-treated BV2 cells was used as conditioned medium (CM). PC12 cells were co-culture with CM for 24 h. Cell viability was determined by MTT assay and cell apoptosis was tested by flow cytometry. BV2 microglia were treated with 10, 20, or 30 μg/ml LPS for 24 h. The expression of TLR4, MyD88, and NF-κB significantly increased. When PC12 cells were co-cultured with CM for 24 h, cell viability decreased. CM up-regulated the Bax level and down-regulated the Bcl-2 protein level in PC12 cells. PC12 cells pretreated with interleukin-1 receptor antagonist (IL-1RA) for 30 min, significantly alleviated CM-induced PC12 cell apoptosis. These results suggest that BV2 microglia activated by LPS triggered TLR4/MyD88/NF-κB signaling pathway that induced the release of IL-1β and could participate in the PC12 cells injury.

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Acknowledgments

This project was supported by grants from the National Natural Science of China (81171041), the Natural Science foundation of Jiangsu Province (BK2011197), Key Subject of Colleges and Universities Natural Science Foundation of Jiangsu Province (13KJA320001), the Natural Science fund for colleges and universities of Jiangsu Province (12KJD320005), the Innovation Project of Nanjing Military Region (ZD13), the University Graduate Student Science and Technology Innovation Project of Jiangsu Province (XYYC-1214), and sponsored by Qing Lan Project.

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The authors have declared that no competing interests exist.

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Authors and Affiliations

  1. Jiangsu Province Key Laboratory of Anesthesiology, Xuzhou Medical College, 209 Tongshan Road, Xuzhou, Jiangsu, 221002, China

    Xiao-jing Dai, Na Li, Le Yu, Zi-yang Chen, Xia Qin & Yong-Mei Zhang

  2. Department of Emergency Medicine, the 97th Hospital of PLA, Xuzhou, Jiangsu, 221002, China

    Rong Hua

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  1. Xiao-jing Dai
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  2. Na Li
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  4. Zi-yang Chen
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Correspondence to Yong-Mei Zhang.

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Xiao-jing Dai and Na Li contributed equally to this work.

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Dai, Xj., Li, N., Yu, L. et al. Activation of BV2 microglia by lipopolysaccharide triggers an inflammatory reaction in PC12 cell apoptosis through a toll-like receptor 4-dependent pathway. Cell Stress and Chaperones 20, 321–331 (2015). https://doi.org/10.1007/s12192-014-0552-1

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