神经酰胺 C16:0激活甲酰肽受体 2 调控棕色脂肪组织产热的转录组学分析

C16:0 ceramide activates formyl peptide receptor 2 to regulate brown adipose tissue thermogenesis: a transcriptomics-based analysis

摘要

目的 探讨神经酰胺 C16:0激活甲酰肽受体 2(formyl peptide receptor 2, FPR2)调控棕色脂肪组织(brown adipose tissue, BAT)产热的下游关键基因及其分子机制。方法 以小鼠 BAT 为研究对象，使用神经酰胺 C16:0 [10 mg/(kg·d)]和空载体连续 3 d 给小鼠进行腹腔注射，之后在 4 ℃环境中暴露 24 h。采用 Western blotting 和 qPCR 技术检测 BAT 中解偶联蛋白 1(uncoupling protein 1, UCP1)的表达水平。利用转录组学测序技术进一步筛选神经酰胺 C16:0调控 BAT 产热的相关基因，并通过 qPCR 法对筛选的基因进行验证。结果 神经酰胺 C16:0可以降低 BAT 中 UCP1 的 mRNA 表达水平[(0.608±0.066) vs. (1.001±0.033), F=3.879, P=0.017]和蛋白水平[(0.725±0.036) vs. (1.000±0.043), F=1.365, P=0.008];转录组学测序共筛选出 909 个差异表达基因(620 个上调，289 个下调)。GO 和 KEGG 富集分析显示，神经酰胺 C16:0显著下调过氧化物酶体增殖物激活受体(peroxisome proliferator-activated receptor, PPAR)和叉头框蛋白 O(forkhead box O, FOXO)信号通路，从中筛选出 7 个基因(SLC27A2、HMGCs2、EHHADH、PCK1、IRS2、SGK2、PLK3)，在 Sox2-CreER^+/-Fpr2^fl/fl小鼠中验证发现，其表达受神经酰胺 C16:0-FPR2 轴特异性调控。结论 神经酰胺 C16:0通过 FPR2 下调小鼠 BAT 中 UCP1 表达，其机制与 PPAR 信号通路和 FOXO 信号通路密切相关，通过调控 SLC27A2、HMGCs2、EHHADH、PCK1、IRS2、SGK2 和 PLK3 基因表达抑制 BAT 产热。

Abstract

Objective To investigate the key downstream genes and molecular mechanisms through which C16:0 ceramide activates formyl peptide receptor 2 (FPR2) to regulate thermogenesis in brown adipose tissue (BAT). Methods Using mouse BAT as the research model, mice were intraperitoneally injected with C16:0 ceramide [10 mg/(kg·d)] or vehicle for three consecutive days, followed by exposure to 4 ℃ for 24 h. The expression level of uncoupling protein 1 (UCP1) in BAT was detected by Western blotting and qPCR. Transcriptome sequencing was further employed to identify genes involved in C16:0 ceramide-mediated regulation of BAT thermogenesis, and the screened genes were validated by qPCR. Results C16:0 ceramide significantly reduced both the mRNA expression level of UCP1 in BAT [(0.608±0.066) vs. (1.001±0.033), F=3.879, P=0.017] and its protein level [(0.725±0.036) vs. (1.000±0.043), F=1.365, P=0.008]. Transcriptome sequencing identified 909 differentially expressed genes (620 up-regulated and 289 down-regulated). GO and KEGG enrichment analyses showed that C16:0 ceramide significantly down-regulated the peroxisome proliferator-activated receptor (PPAR) and forkhead box O (FOXO) signaling pathways, from which seven genes (SLC27A2, HMGCs2, EHHADH, PCK1, IRS2, SGK2, and PLK3) were selected. Validation in Sox2-CreER^+/-Fpr2^fl/fl mice revealed that their expression was specifically regulated by the C16:0 ceramide-FPR2 axis. Conclusion C16:0 ceramide down-regulates UCP1 expression in mouse BAT via FPR2. This mechanism is closely associated with the PPAR and FoxO signaling pathways, and inhibits BAT thermogenesis by regulating the expression of SLC27A2, HMGCs2, EHHADH, PCK1, IRS2, SGK2 and PLK3.

关键词

甲酰肽受体 2 / 神经酰胺 C16:0 / 信号转导通路 / 棕色脂肪组织产热

Key words

Formyl peptide receptor 2 / C16:0 ceramide / Signal transduction pathway / Brown adipose tissue thermogenesis

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基金资助

国家自然科学基金(32130055, 82371629, 82304583)

山东省自然科学基金(ZR2024MC198)