Comas Ferran, Latorre Jèssica, Ortega Francisco, Oliveras-Cañellas Núria, Lluch Aina, Ricart Wifredo, Fernández-Real José Manuel, Moreno-Navarrete José María
Department of Diabetes, Endocrinology and Nutrition, Institut d'Investigació Biomèdica de Girona (IdIBGi), CIBEROBN (CB06/03/010) and Instituto de Salud Carlos III (ISCIII), Girona, Spain.
Department of Diabetes, Endocrinology and Nutrition, Institut d'Investigació Biomèdica de Girona (IdIBGi), CIBEROBN (CB06/03/010) and Instituto de Salud Carlos III (ISCIII), Girona, Spain; Department of Medicine, Universitat de Girona, Girona, Spain.
Redox Biol. 2021 Jun;42:101668. doi: 10.1016/j.redox.2020.101668. Epub 2020 Aug 2.
In the present study, we aimed to investigate the impact of permanent cystathionine-β-Synthase (CBS) gene knockdown in human telomerase reverse transcriptase (hTERT) immortalized human adipose-derived mesenchymal stem cells (ASC52telo) and in their capacity to differentiate into adipocytes. CBS gene KD in ASC52telo cells led to increased cellular inflammation (IL6, CXCL8, TNF) and oxidative stress markers (increased intracellular reactive oxygen species and decreased reduced glutathione levels) in parallel to decreased HS production and rejuvenation (LC3 and SIRT1)-related gene expression. In addition, CBS gene KD in ASC52telo cells resulted in altered mitochondrial respiratory function, characterised by decreased basal respiration (specifically proton leak) and spare respiratory capacity, without significant effects on cell viability and proliferation. In this context, shCBS-ASC52telo cells displayed enhanced adipogenic (FABP4, ADIPOQ, SLC2A4, CEBPA, PPARG)-, lipogenic (FASN, DGAT1)- and adipocyte (LEP, LBP)-related gene expression markers, decreased expression of proinflammatory cytokines, and increased intracellular lipid accumulation during adipocyte differentiation compared to control ASC52telo cells. Otherwise, the increased adipogenic potential of shCBS-ASC52telo cells was detrimental to the ability to differentiate into osteogenic linage. In conclusion, this study demonstrated that permanent CBS gene KD in ASC52telo cells promotes a cellular senescence phenotype with a very increased adipogenic potential, promoting a non-physiological enhanced adipocyte differentiation with excessive lipid storage.
在本研究中，我们旨在探究在人端粒酶逆转录酶（hTERT）永生化的人脂肪来源间充质干细胞（ASC52telo）中永久性胱硫醚-β-合酶（CBS）基因敲低的影响及其分化为脂肪细胞的能力。ASC52telo细胞中的CBS基因敲低导致细胞炎症（IL6、CXCL8、TNF）和氧化应激标志物增加（细胞内活性氧增加，还原型谷胱甘肽水平降低），同时硫酸乙酰肝素生成减少以及与年轻化相关的基因（LC3和SIRT1）表达降低。此外，ASC52telo细胞中的CBS基因敲低导致线粒体呼吸功能改变，其特征为基础呼吸（特别是质子泄漏）和备用呼吸能力降低，对细胞活力和增殖无显著影响。在此背景下，与对照ASC52telo细胞相比，shCBS-ASC52telo细胞在脂肪细胞分化过程中显示出增强的脂肪生成相关基因（FABP4、ADIPOQ、SLC2A4、CEBPA、PPARG）、脂质生成相关基因（FASN、DGAT1）和脂肪细胞相关基因（LEP、LBP）表达标志物，促炎细胞因子表达降低，细胞内脂质积累增加。否则，shCBS-ASC52telo细胞增加的脂肪生成潜能对其分化为成骨谱系的能力有害。总之，本研究表明ASC52telo细胞中永久性CBS基因敲低促进了具有显著增加的脂肪生成潜能的细胞衰老表型，促进了非生理性增强的脂肪细胞分化以及过量脂质储存。
