Objective

To determine the involvement of TNF-α and glycine receptors in the inhibition of pro-inflammatory adipokines in 3T3-L1 cells.

Methods

RT-PCR evidenced glycine receptors in 3T3-L1 adipocytes. 3T3-L1 cells were transfected with siRNA for the glycine (Glrb) and TNF1a (Tnfrsf1a) receptors and confirmed by confocal microscopy. Transfected cells were treated with glycine (10 mM). The expressions of TNF-α and IL-6 mRNA were measured by qRT-PCR, while concentrations were quantified by ELISA.

Results

Glycine decreased the expression and concentration of TNF-α and IL-6; this effect did not occur in the absence of TNF-α receptor due to siRNA. In contrast, glycine produced only slight changes in the expression of TNF-α and IL-6 in the absence of the glycine receptor due to siRNA. A docking analysis confirmed the possibility of binding glycine to the TNF-α1a receptor.

Conclusion

These findings support the idea that glycine could partially inhibit the binding of TNF-α to its receptor and provide clues about the mechanisms by which glycine inhibits the secretion of pro-inflammatory adipokines in adipocytes through the TNF-α receptor.

中文翻译：

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甘氨酸是 TNF 受体的竞争性拮抗剂，介导 3T3-L1 脂肪细胞中炎性细胞因子的表达

客观的

确定 TNF-α 和甘氨酸受体在抑制 3T3-L1 细胞促炎脂肪因子中的作用。

方法

RT-PCR 证实了 3T3-L1 脂肪细胞中的甘氨酸受体。用针对甘氨酸 (Glrb) 和 TNF1a (Tnfrsf1a) 受体的 siRNA 转染 3T3-L1 细胞，并通过共聚焦显微镜进行确认。用甘氨酸 (10 mM) 处理转染的细胞。TNF-α和IL-6 mRNA的表达通过qRT-PCR测量，而浓度通过ELISA定量。

结果

甘氨酸降低TNF-α和IL-6的表达和浓度；由于siRNA，在不存在TNF-α受体的情况下不会发生这种效应。相比之下，在没有甘氨酸受体的情况下，由于siRNA，甘氨酸仅对TNF-α和IL-6的表达产生轻微变化。对接分析证实了甘氨酸与 TNF-α1a 受体结合的可能性。

结论

这些发现支持甘氨酸可以部分抑制 TNF-α 与其受体结合的观点，并提供有关甘氨酸通过 TNF-α 受体抑制脂肪细胞中促炎脂肪因子分泌的机制的线索。