Agareva Margarita, Stafeev Iurii, Michurina Svetlana, Sklyanik Igor, Shestakova Ekaterina, Ratner Elizaveta, Hu Xiang, Menshikov Mikhail, Shestakova Marina, Parfyonova Yelena
Institute of Fine Chemical Technologies Named after M.V. Lomonosov, 119571 Moscow, Russia.
Department of Angiogenesis, National Medical Research Centre of Cardiology Named after Academician E.I. Chazov, 121552 Moscow, Russia.
Life (Basel). 2022 May 6;12(5):688. doi: 10.3390/life12050688.
Sedentary behavior with overnutrition provokes the development of obesity, insulin resistance, and type 2 diabetes mellitus (T2DM). The main progenitor cells of adipose tissue are adipose-derived stem cells (ADSCs) which can change differentiation, metabolic, and secretory phenotypes under obesity conditions. The purpose of this study was to evaluate ADSC osteogenesis activity among patients with obesity in normal glucose tolerance (NGT) and T2DM conditions.
In the study, ADSCs from donors with obesity were used. After clinical characterization, all patients underwent bariatric surgery and ADSCs were isolated from subcutaneous fat biopsies. ADSCs were subjected to osteogenic differentiation, stained with Alizarin Red S, and harvested for real-time PCR and Western blotting. Cell senescence was evaluated with a β-galactosidase-activity-based assay.
Our results demonstrated the significantly increased calcification of ADSC on day 28 of osteogenesis in the T2DM group. These data were confirmed by the statistically significant enhancement of RUNX2 gene expression, which is a master regulator of osteogenesis. Protein expression analysis showed the increased expression of syndecan 1 and collagen I before and during osteogenesis, respectively. Moreover, T2DM ADSCs demonstrated an increased level of cellular senescence.
We suggest that T2DM-associated cellular senescence can cause ADSC differentiation to shift toward osteogenesis, the impaired formation of new fat depots in adipose tissue, and the development of insulin resistance. The balance between ADSC adipo- and osteogenesis commitment is crucial for the determination of the metabolic fate of patients and their adipose tissue.
久坐行为与营养过剩会引发肥胖、胰岛素抵抗和2型糖尿病（T2DM）的发展。脂肪组织的主要祖细胞是脂肪来源干细胞（ADSCs），在肥胖条件下，这些细胞可改变分化、代谢和分泌表型。本研究的目的是评估正常糖耐量（NGT）和T2DM条件下肥胖患者的ADSC成骨活性。
在本研究中，使用了肥胖供体的ADSCs。在进行临床特征分析后，所有患者均接受了减肥手术，并从皮下脂肪活检中分离出ADSCs。将ADSCs进行成骨分化，用茜素红S染色，并收获用于实时PCR和蛋白质印迹分析。通过基于β-半乳糖苷酶活性的检测方法评估细胞衰老情况。
我们的结果表明，在成骨第28天，T2DM组ADSC的钙化显著增加。RUNX2基因表达的统计学显著增强证实了这些数据，RUNX2基因是成骨的主要调节因子。蛋白质表达分析显示，在成骨之前和过程中，syndecan 1和胶原蛋白I的表达分别增加。此外，T2DM的ADSCs表现出细胞衰老水平升高。
我们认为，T2DM相关的细胞衰老可导致ADSC分化向成骨方向转变、脂肪组织中新脂肪库形成受损以及胰岛素抵抗的发展。ADSC脂肪生成和成骨定向之间的平衡对于确定患者及其脂肪组织的代谢命运至关重要。
