Mejia-Garcia Alejandro, Fernandez Geysson Javier, Echeverri Luis Fernando, Balcazar Norman, Acin Sergio
Grupo Genmol. Facultad de Ciencias Exactas y Naturales, Universidad de Antioquia UdeA, Medellín, Colombia.
Grupo Biología y Control de Enfermedades Infecciosas, Universidad de Antioquia UdeA, Medellín, Colombia.
Heliyon. 2024 Jan 10;10(2):e24382. doi: 10.1016/j.heliyon.2024.e24382. eCollection 2024 Jan 30.
Chronic inflammation is crucial in developing insulin resistance and type 2 diabetes. Previous studies have shown that a leaf extract of , with ursolic acid (UA), oleanolic acid (OA), and ursolic acid lactone (UAL) as the main molecules (78 %) mixed with unknown minor metabolites (22 %), provided superior anti-inflammatory, hypoglycemic, and hypolipidemic effects than reconstituted triterpenoid mixtures in macrophage cell lines and a pre-diabetic mouse model. Further identification of the molecular mechanisms of action of this mixture of triterpenes is required. This study aims to analyse the RNA expression profiles of mouse and human macrophage cell lines treated with the natural extract and its components. Activated macrophage cell lines were treated with the natural extract, UA, OA, UAL or a triterpene mixture (M1). RNA was extracted and sequenced using the DNBseq platform and the EnrichR software to perform gene enrichment analysis using the Gene Ontology database, Kyoto Encyclopedia of Genes and Genomes, and Reactome. To conduct clustering analysis, we standardised the normalised counts of each gene and applied k-means clustering. The combination of molecules in the natural extract has an additive or synergic effect that affects the expression of up-regulated genes by macrophage activation. Triterpenes (M1) regulated 76 % of human and 68 % of mouse genes, while uncharacterised minority molecules could regulate 24 % of human and 32 % of mouse genes. The extract inhibited the expression of many cytokines (IL6, IL1, OSM), chemokines (CXCL3), inflammatory mediators (MMP8 and MMP13) and the JAK-STAT signalling pathway in both models. The natural extract has a more powerful immunomodulatory effect than the triterpene mixture, increasing the number of genes regulated in mouse and human models. Our study shows that extract is a promising option for breaking the link between inflammation and insulin resistance.
慢性炎症在胰岛素抵抗和2型糖尿病的发展过程中起着关键作用。先前的研究表明，一种叶提取物，其主要成分（78%）为熊果酸（UA）、齐墩果酸（OA）和熊果酸内酯（UAL），并混合有未知的微量代谢物（22%），在巨噬细胞系和糖尿病前期小鼠模型中，比重组三萜混合物具有更优异的抗炎、降血糖和降血脂作用。需要进一步确定这种三萜混合物的分子作用机制。本研究旨在分析用天然提取物及其成分处理的小鼠和人类巨噬细胞系的RNA表达谱。用天然提取物、UA、OA、UAL或三萜混合物（M1）处理活化的巨噬细胞系。使用DNBseq平台提取RNA并进行测序，并使用EnrichR软件，利用基因本体数据库、京都基因与基因组百科全书和Reactome进行基因富集分析。为了进行聚类分析，我们对每个基因的标准化计数进行了标准化，并应用了k均值聚类。天然提取物中的分子组合具有加性或协同效应，可影响巨噬细胞激活后上调基因的表达。三萜（M1）调节了76%的人类基因和68%的小鼠基因，而未鉴定的微量分子可调节24%的人类基因和32%的小鼠基因。在两种模型中，该提取物均抑制了许多细胞因子（IL6、IL1、OSM）、趋化因子（CXCL3）、炎症介质（MMP8和MMP13）以及JAK-STAT信号通路的表达。天然提取物比三萜混合物具有更强的免疫调节作用，增加了在小鼠和人类模型中受调节的基因数量。我们的研究表明，该提取物是打破炎症与胰岛素抵抗之间联系的一个有前景的选择。
