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D-beta-hydroxybutyrate protects against microglial activation in lipopolysaccharide-treated mice and BV-2 cells

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Abstract

Microglial activation is a key event in neuroinflammation, which, in turn, is a central process in neurological disorders. In this study, we investigated the protective effects of D-beta-hydroxybutyrate (BHB) against microglial activation in lipopolysaccharide (LPS)-treated mice and BV-2 cells. The effects of BHB in mice were assessed using behavioral testing, morphological analysis and immunofluorescence labeling for the microglial marker ionizing calcium-binding adaptor molecule 1 (IBA-1) and the inflammatory cytokine interleukin-6 (IL-6) in the hippocampus. Moreover, we examined the levels of the inflammatory IL-6 and tumor necrosis factor-α (TNF-α), as well as those of the neuroprotective brain-derived neurotrophic factor (BDNF) and transforming growth factor-β (TGF-β) in the brain. In addition, we examined the effects of BHB on IL-6, TNF-α, BDNF, TGF-β, reactive oxygen species (ROS) level and cell viability in LPS-stimulated BV-2 cells. BHB treatments attenuated behavioral abnormalities, reduced the number of IBA-1-positive cells and the intensity of IL-6 fluorescence in the hippocampus, with amelioration of microglia morphological changes in the LPS-treated mice. Furthermore, BHB inhibited IL-6 and TNF-α generation, but promoted BDNF and TGF-β production in the brain of LPS-treated mice. In vitro, BHB inhibited IL-6 and TNF-α generation, increased BDNF and TGF-β production, reduced ROS level, ameliorated morphological changes and elevated cell viability of LPS-stimulated BV-2 cells. Together, our findings suggest that BHB exerts protective effects against microglial activation in vitro and in vivo, thereby reducing neuroinflammation.

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Acknowledgements

This study was supported by grants from the National Natural Science Foundation of China (82170846) and the Natural Science Foundation of Hebei (H2020206386 and H2020206328). The authors thank Professor Haishui Shi (Neuroscience Research Center, Institute of Medical and Health Science, Hebei Medical University, Hebei, China) for guidance of behavioral test. We thank Barry Patel, PhD, from Liwen Bianji (Edanz) (www.liwenbianji.cn/), for editing the English text of a draft of this manuscript.

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  1. Yuping Zhang and Kun Liu contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology, Hebei Key Lab of Laboratory Animal Science, Hebei Medical University, No. 361 East Zhongshan Road, Shijiazhuang, 050017, Hebei, People’s Republic of China

    Yuping Zhang, Yunpeng Li, Yujie Ma, Yu Wang & Yanning Li

  2. Department of Biochemistry, College of Integrated Chinese and Western Medicine, Hebei Medical University, No. 361 East Zhongshan Road, Shijiazhuang, 050017, Hebei, People’s Republic of China

    Kun Liu, Zihan Fan & Jinsheng Qi

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  1. Yuping Zhang
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Contributions

Yanning Li designed the study, reviewed and edited the manuscript. Yuping Zhang and Kun Liu performed experiments and analyzed the data. Yunpeng Li and Yujie Ma performed experiments and wrote the manuscript. Yu Wang and Zihan Fan performed experiments. Jinsheng Qi discussed the results and commented on the manuscript. All authors read and approved the final manuscript.

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Supplementary Figure 1

Effects of BHB on microglial IL-6 in the hippocampus of LPS-treated mice. (A) Immunofluorescence of IL-6 (green) and microglial marker IBA-1 (red) was shown to reflect microglial IL-6. Scale bar, 10 μm. (B) The relative intensity of IL-6 fluorescence was calculated (n = 3). Data were means ± SD. * p < 0.05 versus Con; # p < 0.05 versus LPS; analyzed by one-way ANOVA and least significant difference test. (PNG 292 kb)

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Zhang, Y., Liu, K., Li, Y. et al. D-beta-hydroxybutyrate protects against microglial activation in lipopolysaccharide-treated mice and BV-2 cells. Metab Brain Dis 38, 1115–1126 (2023). https://doi.org/10.1007/s11011-022-01146-7

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