INTRODUCTION Mesenchymal stem cells (MSCs) have endogenous reparative properties, and may constitute an exogenous therapeutic intervention in patients with chronic kidney disease. The microenvironment of metabolic syndrome (MetS) induces fat inflammation, with abundant expression of tumor necrosis factor (TNF)-α. MetS may also alter the content of adipose tissue-derived MSCs, and we hypothesized that the inflammatory profile of MetS manifests via upregulating MSC mRNAs and proteins of the TNF-α pathway. METHODS Domestic pigs were fed a 16-week Lean or MetS diet (n = 4 each). MSCs were harvested from abdominal subcutaneous fat, and their extracellular vesicles (EVs) isolated. Expression profiles of mRNAs and proteins in MSCs and EVs were obtained by high-throughput sequencing and proteomics. Nuclear translocation of the pro-inflammatory transcription factor (NF)-kB was evaluated in MSC and in pig renal tubular cells (TEC) co-incubated with EVs. RESULTS We found 13 mRNAs and 4 proteins in the TNF-α pathway upregulated in MetS- vs. Lean-MSCs (fold-change > 1.4, p < 0.05), mostly via TNF-α receptor-1 (TNF-R1) signaling. Three mRNAs were upregulated in MetS-EVs. MetS-MSCs, as well as TECs co-incubated with MetS-EVs, showed increased nuclear translocation of NF-kB. Using qPCR, JUNB, MAP2K7 and TRAF2 genes followed the same direction of RNA-sequencing findings. CONCLUSIONS MetS upregulates the TNF-α transcriptome and proteome in swine adipose tissue-derived MSCs, which are partly transmitted to their EV progeny, and are associated with activation of NF-kB in target cells. Hence, the MetS milieu may affect the profile of endogenous MSCs and their paracrine vectors and limit their use as an exogenous regenerative therapy. Anti-inflammatory strategies targeting the TNF-α pathway might be a novel strategy to restore MSC phenotype, and in turn function.

中文翻译：

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代谢综合征猪脂肪组织来源的间充质干细胞中肿瘤坏死因子-α转录组和蛋白质组上调


简介 间充质干细胞（MSC）具有内源性修复特性，可能构成慢性肾病患者的外源性治疗干预措施。代谢综合征（MetS）的微环境会诱导脂肪炎症，并大量表达肿瘤坏死因子（TNF）-α。 MetS 还可能改变脂肪组织来源的 MSC 的含量，我们假设 MetS 的炎症特征通过上调 MSC mRNA 和 TNF-α 途径的蛋白来表现。方法 家猪饲喂 16 周的瘦肉或 MetS 饮食（每只 n = 4）。从腹部皮下脂肪中收获 MSC，并分离其细胞外囊泡 (EV)。通过高通量测序和蛋白质组学获得了 MSC 和 EV 中 mRNA 和蛋白质的表达谱。在 MSC 和与 EV 共孵育的猪肾小管细胞 (TEC) 中评估了促炎转录因子 (NF)-kB 的核转位。结果我们发现，在 MetS- 与 Lean-MSC 中，TNF-α 通路中的 13 个 mRNA 和 4 个蛋白上调（倍数变化 > 1.4，p < 0.05），主要是通过 TNF-α 受体-1 (TNF-R1) 信号传导。 MetS-EV 中的 3 个 mRNA 上调。 MetS-MSC 以及与 MetS-EV 共孵育的 TEC 均表现出 NF-kB 核转位增加。使用 qPCR，JUNB、MAP2K7 和 TRAF2 基因遵循与 RNA 测序结果相同的方向。结论 MetS 上调猪脂肪组织来源的 MSC 中的 TNF-α 转录组和蛋白质组，这些转录组和蛋白质组部分传递给其 EV 后代，并且与靶细胞中 NF-kB 的激活相关。因此，MetS 环境可能会影响内源性 MSC 及其旁分泌载体的特征，并限制它们作为外源性再生疗法的用途。 针对 TNF-α 通路的抗炎策略可能是恢复 MSC 表型并进而恢复功能的新策略。