Antiadipogenic and antiobesogenic effects of pterostilbene in 3T3-L1 preadipocyte models.

Birgül Gülnar, İpek Canatar, Sibel Özdaş
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Abstract

Since obesity causes at least 2.8 million death each year and is a major risk factor for many diseases, it is critical to evaluate alternative treatment approaches. In this context, studies on the research of natural product-based therapeutics in the fight against obesity are increasing. In this study, it was aimed to evaluate the antiadipogenic and antiobesogenic efficacy of pterostilbene a natural phenolic compound in 3T3-L1 cells. The mature 3T3-L1 adipocytes were exposed to pterostilbene at different concentrations and half-maximum inhibitory concentrations (IC50) were determined by MTT assay. Oil-Red-O staining was applied to determine lipid accumulation. Phase contrast microscopy, Giemsa, F-actin and DAPI staining were applied to examine the efficacy of pterostilbene on the morphology of 3T3-L1 adipocyte cells. Moreover, expressions of adinopectin and glucose transporter-4 (Glut-4) in relation to insulin resistance were evaluated using immunofluorescent staining and qRT-PCR. Pterostilbene caused no significant cytotoxicity towards preadipocytes at concentrations ≤7.5 -M and the percentage of viable cells remained above about 86% for 24 h, 48 h and 72 h (p > 0.05). Therefore, pterostilbene treatment at 5 and 7.5 -M was used in the subsequent experiments as safe dosages. In addition, it was observed that pterostilbene treatment reduced lipid accumulation in adipocyte differentiation. Adipocytes treated with a dose of 7.5 -M for 14 days showed less intense lipid deposition and a more spindle-like morphology compared to the adipocytes treated with a dose of 5 -M. Especially on the 14th day, actin filaments were filamentous in adipocytes treated with pterostilbene 7.5 -M compared to the adipocytes treated with a dose of 5 -M; the filaments were similarly oriented as in preadipocytes, and chromatin condensation was observed to be quite high. Our data suggests that the pterostilbene supplementation may help weight control and the antiadipogenic and that antiobesogenic activity is mediated in part by reduction of lipid accumulation and induction of Glut-4 and Adiponectin levels.

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紫檀芪在3T3-L1前脂肪细胞模型中的抗脂肪生成和抗肥胖作用。
由于肥胖每年至少造成280万人死亡,并且是许多疾病的主要危险因素,因此评估替代治疗方法至关重要。在此背景下,研究以天然产品为基础的治疗方法来对抗肥胖的研究正在增加。本研究旨在评价天然酚类化合物紫檀芪对3T3-L1细胞的抗脂肪和抗肥胖作用。将成熟的3T3-L1脂肪细胞暴露于不同浓度的紫檀芪中,采用MTT法测定半最大抑制浓度(IC50)。油-红- o染色测定脂质积累。采用相差显微镜、Giemsa、F-actin和DAPI染色检测紫檀芪对3T3-L1脂肪细胞形态的影响。此外,采用免疫荧光染色和qRT-PCR方法评估adinopectin和glucose transporter-4 (Glut-4)的表达与胰岛素抵抗的关系。在浓度≤7.5 -M时,紫檀芪对前脂肪细胞无明显的细胞毒性作用,24 h、48 h和72 h活细胞率均保持在86%以上(p > 0.05)。因此,在后续实验中,采用5和7.5 -M紫檀芪处理作为安全剂量。此外,还观察到紫檀芪处理可减少脂肪细胞分化过程中的脂质积累。与5 -M剂量处理的脂肪细胞相比,7.5 -M剂量处理14天的脂肪细胞显示出较少的脂质沉积和更多的纺锤状形态。特别是在第14天,与5 -M剂量的脂肪细胞相比,7.5 -M紫檀芪处理的脂肪细胞的肌动蛋白丝呈丝状;纤维的方向与前脂肪细胞相似,染色质浓缩程度很高。我们的数据表明,补充紫芪可能有助于控制体重和抗脂肪生成,而抗肥胖活性部分是通过减少脂质积累和诱导Glut-4和脂联素水平介导的。
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