Atractylodes lancea (Thunb.) DC. [Asteraceae] rhizome-derived exosome-like nanoparticles suppress lipopolysaccharide-induced inflammation in murine microglial cells

Kei Kawada; Tomoaki Ishida; Shumpei Morisawa; Kohei Jobu; Youichirou Higashi; Fuka Aizawa; Kenta Yagi; Yuki Izawa-Ishizawa; Takahiro Niimura; Shinji Abe; Mitsuhiro Goda; Mitsuhiko Miyamura; Keisuke Ishizawa

doi:10.3389/fphar.2024.1302055

Atractylodes lancea (Thunb.) DC. [Asteraceae] rhizome-derived exosome-like nanoparticles suppress lipopolysaccharide-induced inflammation in murine microglial cells

Front Pharmacol. 2024 Apr 26:15:1302055. doi: 10.3389/fphar.2024.1302055. eCollection 2024.

Authors

Kei Kawada^{1

2}, Tomoaki Ishida³, Shumpei Morisawa³, Kohei Jobu³, Youichirou Higashi⁴, Fuka Aizawa^{1

5}, Kenta Yagi^{1

6}, Yuki Izawa-Ishizawa^{1

7}, Takahiro Niimura^{1

6}, Shinji Abe², Mitsuhiro Goda^{1

5}, Mitsuhiko Miyamura⁸, Keisuke Ishizawa^{1

6

7}

Affiliations

¹ Department of Clinical Pharmacology and Therapeutics, Tokushima University Graduate School of Biomedical Sciences, Tokushima, Japan.
² Department of Clinical Pharmacy Practice Pedagogy, Tokushima University Graduate School of Biomedical Sciences, Tokushima, Japan.
³ Department of Pharmacy, Kochi Medical School Hospital, Kochi, Japan.
⁴ Department of Pharmacology, Kochi Medical School, Kochi University, Kochi, Japan.
⁵ Department of Pharmacy, Tokushima University Hospital, Tokushima, Japan.
⁶ Clinical Research Center for Developmental Therapeutics, Tokushima University Hospital, Tokushima, Japan.
⁷ Department of General Medicine, Taoka Hospital, Tokushima, Japan.
⁸ Center for Regional Sustainability and Innovation, Kochi University, Kochi, Japan.

Abstract

Background: Exosome-like nanoparticles (ELNs) mediate interspecies intercellular communications and modulate gene expression.

Hypothesis/purpose: In this study, we isolated and purified ELNs from the dried rhizome of Atractylodes lancea (Thunb.) DC. [Asteraceae] (ALR-ELNs), a traditional natural medicine, and investigated their potential as neuroinflammatory therapeutic agents.

Methods: ALR-ELN samples were isolated and purified using differential centrifugation, and their physical features and microRNA contents were analyzed through transmission electron microscopy and RNA sequencing, respectively. BV-2 microglial murine cells and primary mouse microglial cells were cultured in vitro, and their ability to uptake ALR-ELNs was explored using fluorescence microscopy. The capacity of ALR-ELNs to modulate the anti-inflammatory responses of these cells to lipopolysaccharide (LPS) exposure was assessed through mRNA and protein expression analyses.

Results: Overall, BV-2 cells were found to internalize ALR-ELNs, which comprised three microRNAs (ath-miR166f, ath-miR162a-5p, and ath-miR162b-5p) that could have anti-inflammatory activity. Pretreatment of BV-2 cells with ALR-ELN prevented the pro-inflammatory effects of LPS stimulation by significantly reducing the levels of nitric oxide, interleukin-1β, interleukin-6, and tumor necrosis factor-α. Notably, the mRNA levels of Il1b, Il6, iNos, ccl2, and cxcl10 in BV-2 cells, which increased upon LPS exposure, were significantly reduced following ALR-ELN treatment. Moreover, the mRNA levels of heme oxygenase 1, Irf7, ccl12, and Irg1 also increased significantly following ALR-ELN treatment. In addition, pretreatment of primary mouse microglial cells with ALR-ELN prevented the pro-inflammatory effects of LPS stimulation by significantly reducing the levels of nitric oxide.

Conclusion: Our findings indicate that ALR-ELNs exhibit anti-inflammatory effects on murine microglial cells. Further validation may prove ALR-ELNs as a promising neuroinflammatory therapeutic agent.

Keywords: Atractylodes lancea (Thunb.) DC. [Asteraceae] rhizome; Kampo medicine; exosome-like nanoparticle; microRNA; microglia; neuroinflammation.

Grants and funding

The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This work was supported by the KAKENHI program of the Japan Society for the Promotion of Science (grant number 22K15304). The funding agencies had no role in the collection, analysis, and interpretation of data, in the writing of the report, or in the decision to submit the article for publication.