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FOXO3a regulates lipid accumulation and adipocyte inflammation in adipocytes through autophagy
Inflammation Research ( IF 6.7 ) Pub Date : 2021-04-23 , DOI: 10.1007/s00011-021-01463-0
Xiaoyan Zhang 1 , Qiang Liu 2 , Xuane Zhang 1 , Kai Guo 1 , Xuelian Zhang 1 , Zunhai Zhou 1
Affiliation  

Background

FOXO3a is a widely studied transcription factor and plays an important role in a variety of biology. The purpose of this study was to explore the role and potential mechanism of FOXO3a on lipid accumulation and adipocyte inflammation in adipocytes through regulation of autophagy.

Methods

The obese mouse model was successfully induced by high-fat diet. SiRNA targeting FOXO3a was transfected into differentiation of 3T3-L1 adipocytes to reduce the expression of FOXO3a. The culture medium of RAW264.7 cells was added to the differentiated 3T3-L1 adipocytes to form a co-culture system. Subsequently, ELISA or AdipoRed assay was performed to measure the expression of triglyceride (TG) and cholesterol (TC) in mouse adipose tissue or differentiation of 3T3-L1 adipocytes. Adipocyte differentiation was detected by Oil Red O-staining. Ad-mCherry-GFP-LC3II was used to detect the level of autophagy in differentiation of 3T3-L1 adipocytes. Western blotting or qRT-PCR was used to detect the expression of FOXO3a, autophagy-related proteins (beclin 1, CEBPβ, PPARγ, ACC1 and KLF4), inflammatory cytokines (TNF-α, IL-1β, IL-6 and MCP1), NF-κB signal pathway-related proteins or adipokines (Adiponectin, AdipoR1 and resistin) in differentiated 3T3-L1 or RAW264.7 cells.

Results

The expression of FOXO3a and autophagy levels were significantly increased in visceral adipose tissue of obese mice and differentiation of 3T3-L1 adipocytes. Downregulation of FOXO3a significantly inhibited the autophagy and lipid accumulation in differentiation of 3T3-L1 adipocytes. In addition, FOXO3a knockdown significantly reduced Lipopolysaccharide (LPS)-induced inflammation and adipokines release in RAW264.7 cells treated with the culture medium of 3T3-L1 adipocytes. These above activity changes could be reversed by autophagy inducer rapamycin.

Conclusion

FOXO3a could promote lipid accumulation and inflammation in differentiated 3T3-L1 adipocytes by targeting autophagy. Our results provide a new theoretical basis for FOXO3a to regulate obesity.



中文翻译:

FOXO3a通过自噬调节脂肪细胞中的脂质积累和脂肪细胞炎症

背景

FOXO3a 是一种被广泛研究的转录因子,在多种生物学中发挥着重要作用。本研究的目的是探讨 FOXO3a 通过调节自噬对脂肪细胞中脂质积累和脂肪细胞炎症的作用和潜在机制。

方法

高脂饮食成功诱导肥胖小鼠模型。将靶向 FOXO3a 的 SiRNA 转染到分化的 3T3-L1 脂肪细胞中以降低 FOXO3a 的表达。将RAW264.7细胞培养基加入分化的3T3-L1脂肪细胞,形成共培养体系。随后,进行ELISA或AdipoRed测定以测量小鼠脂肪组织中甘油三酯(TG)和胆固醇(TC)的表达或3T3-L1脂肪细胞的分化。通过油红O染色检测脂肪细胞分化。Ad-mCherry-GFP-LC3II 用于检测 3T3-L1 脂肪细胞分化中的自噬水平。Western blotting 或 qRT-PCR 检测 FOXO3a、自噬相关蛋白(beclin 1、CEBPβ、PPARγ、ACC1 和 KLF4)、炎性细胞因子(TNF-α、IL-1β、IL-6 和 MCP1)的表达,

结果

FOXO3a的表达和自噬水平在肥胖小鼠内脏脂肪组织和3T3-L1脂肪细胞分化中显着增加。FOXO3a 的下调显着抑制了 3T3-L1 脂肪细胞分化中的自噬和脂质积累。此外,在用 3T3-L1 脂肪细胞培养基处理的 RAW264.7 细胞中,FOXO3a 敲低显着降低了脂多糖 (LPS) 诱导的炎症和脂肪因子释放。自噬诱导剂雷帕霉素可以逆转上述活性变化。

结论

FOXO3a 可以通过靶向自噬促进分化的 3T3-L1 脂肪细胞中的脂质积累和炎症。我们的研究结果为 FOXO3a 调节肥胖提供了新的理论基础。

更新日期:2021-04-23
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