Berberine increases expression of GATA-2 and GATA-3 during inhibition of adipocyte differentiation

• Published in: Phytomedicine (Stuttgart) 2009-09, Vol.16 (9), p.864-873
• Main Authors: Hu, Y., Davies, G.E.
• Format: Article
• Language: English
• Subjects: 3T3-L1; 3T3-L1 Cells; Adipocyte differentiation; Adipocytes - drug effects; Adipogenesis - drug effects; Animals; Berberine; Berberine - chemistry; Berberine - pharmacology; C/EBPα; CCAAT-Enhancer-Binding Protein-alpha - genetics; CCAAT-Enhancer-Binding Protein-alpha - metabolism; Cell differentiation; Cell Line; Control; Down-Regulation; Fat cells; GATA; GATA2 Transcription Factor - genetics; GATA2 Transcription Factor - metabolism; GATA3 Transcription Factor - genetics; GATA3 Transcription Factor - metabolism; Gene Expression - drug effects; Mice; Physiological aspects; Plant Extracts - pharmacology; PPAR gamma - genetics; PPAR gamma - metabolism; PPARγ; RNA, Messenger - metabolism
• ISSN: 0944-7113; 1618-095X
• DOI: 10.1016/j.phymed.2009.03.002

It is known that a number of transcription factors are key regulators in the complex process of adipocyte differentiation including peroxisome proliferator activated receptor γ (PPARγ) and the CCAAT enhancer binding protein α (C/EBPα). Studies have demonstrated that in pre-adipocyte 3T3-L1 cells constitutive expression of the DNA binding proteins GATA-2 and GATA-3 results in protein/protein interactions with C/EBPα resulting in down regulation of PPARγ and subsequent suppressed adipocyte differentiation with cells trapped at the pre-adipocyte stage. Thus it appears that GATA-2 and GATA-3 are of critical importance in regulating adipocyte differentiation through molecular interactions with PPARγ and C/EBPα. Recent reports suggest that berberine, an isoquinoline derivative alkaloid isolated from many medicinal herbs prevents differentiation of 3T3-L1 cells via a down regulation of PPARγ and C/EBPα expression. The aim of this study was to determine the effect of berberine on GATA-2 and 3 gene and protein expression levels during differentiation of 3T3-L1 cells. MTT (Methylthiazolyldiphenyl-tetrazolium bromide) was used to detect the cytotoxic effects of berberine on the viability of 3T3-L1 cells during proliferation and differentiation. Differentiation of 3T3-L1 cells was monitored by Oil Red O staining and RT-PCR of PPARγ and C/EBPα and the expression of GATA-2 and 3 was determined by RT-PCR and Western Blot. Results show that following treatment with 8 μM berberine the mRNA and protein expression levels of GATA-2 and 3 were elevated and accompanied by inhibited adipocyte differentiation. These results may lead to the use of berberine to target the induction of specific genes such as GATA-2 and GATA-3 which affect adipocyte differentiation.