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TLR4 signaling is involved in the protective effect of propofol in BV2 microglia against OGD/reoxygenation

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Abstract

Propofol exhibits neuroprotective effects against hypoxic–ischemic brain injury, but the underlying mechanisms are still not clear. Toll-like receptor 4 (TLR4) plays a considerable role in the induction of innate immune and inflammatory responses. The purposes of this study are to investigate the effect of propofol on the oxygen and glucose deprivation (OGD)/reoxygenation (OGD/R) BV2 microglia and to explore the role of TLR4/myeloid differentiation protein 88 (MyD88)/nuclear factor-kappa B (NF-κB) pathway in the neuroprotective effects of propofol. BV2 microglia were placed into an airtight chamber and in glucose-free medium for OGD/reoxygenation. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay. TLR4 and its downstream signaling molecules, MyD88 and NF-κB expressions were detected by Western blotting. Level of tumor necrosis factor alpha (TNF-α) in culture medium was determined with enzyme-linked immunosorbent assay. BV2 microglia apoptosis was determined by flow cytometry. We found that pretreatment with propofol significantly alleviated the hypoxic injury in BV2 microglia. Propofol inhibited upregulation of TLR4, MyD88, and NF-κB expressions in BV2 microglia exposed to OGD/reoxygenation. Propofol pretreatment also significantly reduced the production of TNF-α and apoptosis in OGD/reoxygenation BV2 microglia. The results indicated that TLR4 and its downstream MyD88-dependent signaling pathway contributed to neuroprotection of propofol to microglia exposed to OGD/reoxygenation.

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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), the Natural Science fund for colleges and universities of Jiangsu Province (12KJD320005), the Social Development Science and Technology Project of Xuzhou city (xzzd1051), Qing Lan Project, the University Graduate Student Science and Technology Innovation Project of Jiangsu Province (cx2211-0752, xyyc-12214), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions of Jiangsu Province.

Competing interests

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, 221004, China

    Xia Qin, Zhen-Quan Sun, Xue-Wei Zhang, Xiao-Jing Dai, Shan-Shan Mao & Yong-Mei Zhang

  2. Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application Technology, Xuzhou, China

    Xia Qin, Zhen-Quan Sun, Xue-Wei Zhang, Xiao-Jing Dai, Shan-Shan Mao & Yong-Mei Zhang

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  1. Xia Qin
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Correspondence to Yong-Mei Zhang.

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X. Qin and Z.-Q. Sun contributed equally to this work.

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Fig. S1

No dramatic change occurred when propofol was added into BV2 microglia. 100 μM propofol was added into untreated control BV2 microglial cells, and cell viability was assessed using the MTT assay 15 h later. The results are expressed as MTT OD values. Each value indicates the mean ± SD and is representative of results obtained from four independent experiments. (JPEG 19 kb)

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Fig. S2

Propofol had no significant effect on control BV2 microglia in TLR4 expression. Twenty-five micromolars propofol was added into untreated control BV2 microglial cells, and TLR4 expression was assessed with Western blotting. a Cell lysates were immunoblotted with TLR4 subsequently reprobed with β-actin. These results are representative for two independent experiments. b Densitometric analysis of protein expression in cells with Image J. (JPEG 29 kb)

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Qin, X., Sun, ZQ., Zhang, XW. et al. TLR4 signaling is involved in the protective effect of propofol in BV2 microglia against OGD/reoxygenation. J Physiol Biochem 69, 707–718 (2013). https://doi.org/10.1007/s13105-013-0247-6

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  • DOI: https://doi.org/10.1007/s13105-013-0247-6

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