Dataset Information

Docosahexaenoic acid attenuates metabolic dysfunctions induced by lipopolysaccharide in BV-2 microglial cells

ABSTRACT: Microglial cell activation has been linked to many neurodegenerative diseases. Upon stimulation by lipopolysaccharide (LPS), a number of proteins involved in inflammatory and oxidative pathways are activated. Production of nitric oxide has been regarded as a signature marker of inflammatory responses. Our recent studies demonstrated the effects of docosahexaenoic acid (DHA) to inhibit the LPS-induced inflammatory responses in BV-2 microglial cells. DHA also can upregulate the anti-oxidative pathway involving nuclear factor erythroid 2-Like 2 (Nrf2) and synthesis of heme oxygenase-1 (HO-1), a potent anti-oxidative enzyme. In order to further understand the proteins involved, this study used a label-free quantitative proteomics approach to examine effects of DHA and LPS on proteins and signaling pathways in microglial cells.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Microglial Cell

SUBMITTER: Bo Yang

LAB HEAD: C. Michael Greenlief

PROVIDER: PXD013086 | Pride | 2020-04-20

REPOSITORIES: Pride

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Publications

Quantitative Proteomics Reveals Docosahexaenoic Acid-Mediated Neuroprotective Effects in Lipopolysaccharide-Stimulated Microglial Cells.

Yang Bo B Li Runting R Liu Pei N PN Geng Xue X Mooney Brian P BP Chen Chen C Cheng Jianlin J Fritsche Kevin L KL Beversdorf David Q DQ Lee James C JC Sun Grace Y GY Greenlief C Michael CM

Journal of proteome research 20200427 6

The high levels of docosahexaenoic acid (DHA) in cell membranes within the brain have led to a number of studies exploring its function. These studies have shown that DHA can reduce inflammatory responses in microglial cells. However, the method of action is poorly understood. Here, we report the effects of DHA on microglial cells stimulated with lipopolysaccharides (LPSs). Data were acquired using the parallel accumulation serial fragmentation method in a hybrid trapped ion mobility-quadrupole ...[more]

PMID: 32302149

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Project description:Background: Docosahexaenoic acid (DHA) is a natural compound with anticancer and anti-angiogenesis activity that is currently under investigation as both a preventative agent and an adjuvant to breast cancer therapy. However, the precise mechanisms of DHA’s anticancer activities are unclear. It is understood that the intercommunication between cancer cells and their microenvironment is essential to tumor angiogenesis. Exosomes are extracellular vesicles that are important mediators of intercellular communication and play a role in promoting angiogenesis. However, very little is known about the contribution of breast cancer exosomes to tumor angiogenesis or whether exosomes can mediate DHA’s anticancer action. Results: Exosomes were collected from MCF7 and MDA-MB-231 breast cancer cells after treatment with DHA. We observed an increase in exosome secretion and exosome microRNA contents from the DHA-treated cells. The expression of 83 microRNAs in the MCF7 exosomes was altered by DHA (>2-fold). The most abundant exosome microRNAs (let-7a, miR-23b, miR-27a/b, miR-21, let-7, and miR-320b) are known to have anti-cancer and/or anti-angiogenic activity. These microRNAs were also increased by DHA treatment in the exosomes from other breast cancer lines (MDA-MB-231, ZR751 and BT20), but not in exosomes from normal breast cells (MCF10A). When DHA-treated MCF7 cells were co-cultured with or their exosomes were directly applied to endothelial cell cultures, we observed an increase in the expression of these microRNAs in the endothelial cells. Furthermore, overexpression of miR-23b and miR-320b in endothelial cells decreased the expression of their pro-angiogenic target genes (PLAU, AMOTL1, NRP1 and ETS2) and significantly inhibited tube formation by endothelial cells, suggesting that the microRNAs transferred by exosomes mediate DHA’s anti-angiogenic action. These effects could be reversed by knockdown of the Rab GTPase, Rab27A, which controls exosome release. Conclusions: We conclude that DHA alters breast cancer exosome secretion and microRNA contents, which leads to the inhibition of angiogenesis. Our data demonstrate that breast cancer exosome signaling can be targeted to inhibit tumor angiogenesis and provide new insight into DHA’s anticancer action, further supporting its use in cancer therapy. Examination of small RNA populations in MCF7 cells and exosomes after DHA treatment.

Project description:Preclinical and clinical studies suggest that consumption of long chain omega-3 polyunsaturated fatty acids (PUFAs) reduces severity chronic and autoimmune diseases. These ameliorative effects have been conventionally linked in part to downregulation of expression of proinflammatory cytokine and chemokine genes and more recently, associated with inhibition of Type 1 interferon (IFN1)-regulated gene expression. Here we used single cell RNA sequencing (scRNAseq) to gain new mechanistic perspectives of how the omega-3 PUFA docosahexaenoic acid (DHA) influences TLR4-driven proinflammatory and IFN1-regulated gene expression in a novel self-renewing murine fetal liver-derived macrophage (FLM) model. FLMs were cultured with 25 µM DHA or vehicle for 24 h, treated with modest concentration of LPS (20 ng/ml) for 1 and 4 h, and then subjected to scRNAseq using 10X Chromium System. At 0 h (i.e., in the absence of LPS), DHA increased expression of genes associated with the NRF2 antioxidant response (Sqstm1, Hmox1, Chchd10) and metal homeostasis (Mt1, Mt2, Ftl1, Fth1) suggesting cells are polarized towards a more anti-inflammatory phenotype. At 1 h post-treatment, DHA inhibited LPS-induced cholesterol synthesis genes (Scd1, Scd2, Pmvk, Cyp51, Hmgcs1, and Fdps) which potentially could contribute to interference with TLR4-mediated inflammatory signaling. At 4 h post-treatment, LPS-treated FLMs reflected a robust proinflammatory response including upregulation of cytokine (Il1a, Il1b, Tnf) and chemokine (Ccl2, Ccl3, Ccl4, Ccl7) genes as well as IFN1-regulated (Irf7, Mx1, Oasl1, Ifit1) genes, many of which were suppressed by DHA. Using SCENIC analysis to identify gene expression networks, we found DHA modestly downregulated LPS-induced expression of NF-κB-target genes. Moreover, LPS induced a subset of FLMs simultaneously expressing NF-κB- and IRF7/STAT1/STAT2-target genes that were conspicuously absent in DHA-pretreated FLMs, suggesting that DHA more potently targets the IFN1 response over the NF-kB response. Altogether, scRNAseq generated a valuable dataset that provides new insights into multiple overlapping mechanisms by which DHA may transcriptionally or post-transcriptionally regulate LPS-induced proinflammatory and IFN1-driven responses in macrophages

Dataset Information

Docosahexaenoic acid attenuates metabolic dysfunctions induced by lipopolysaccharide in BV-2 microglial cells

Publications

Quantitative Proteomics Reveals Docosahexaenoic Acid-Mediated Neuroprotective Effects in Lipopolysaccharide-Stimulated Microglial Cells.

Similar Datasets

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