Biochanin A induces a brown‐fat phenotype via improvement of mitochondrial biogenesis and activation ofAMPKsignaling in murineC3H10T1/2 mesenchymal stem cells

Abstract

In this study, we investigated the effect of Biochanin A (BioA), an O‐methylated isoflavone on the brown‐fat phenotype formation and on the associated thermogenic program including mitochondrial biogenesis and lipolysis in C3H10T1/2 MSCs. Our data demonstrates that Treatment with BioA in an adipogenic differentiation cocktail induced formation of brown‐fat–like adipocytes from C3H10T1/2 MSCs without treatment with a known browning inducer (rosiglitazone or T3) at an early stage of differentiation. The formation of brown‐fat–like adipocytes by BioA treatment was evidenced by upregulation of key thermogenic markers: Ucp1, Pgc1α, Prdm16, and Pparγ. BioA also increased the expression of beige (Cd137 and Fgf21) and brown (Elovl3 and Zic1)‐specific markers. Additionally, BioA treatment promoted mitochondrial biogenesis, judging by the upregulation of genes; Cox8b, Cidea, Dio2, Sirt1, Opa1, and Fis1. BioA treatment increased the amount of mitochondrial DNA and its encoded proteins: oxidative phosphorylation complexes (I–V); this change was associated with high oxygen consumption by C3H10T1/2 MSCs. A small‐interfering‐RNA–induced gene knockdown and experiments with dorsomorphin‐driven competitive inhibition revealed that BioA exerts the thermogenic action via activation of AMPK signaling. Our study shows the mechanism of BioA‐induced promotion of a brown‐fat phenotype. Nonetheless, clinical research is necessary to validate BioA as a brown‐fat‐like signature inducer.

Hepatic PTEN Signaling Regulates Systemic Metabolic Homeostasis through Hepatokines-Mediated Liver-to-Peripheral Organs Crosstalk

“…The fact that Angptl4 expression is induced by anti-diabetic drugs further fuels the controversy about the role of ANGPTL4 in metabolic homeostasis as discussed earlier. Finally, Lect2 expression was not found to be significantly sensitive to anti-diabetic drugs and phytoestrogens, while Fgf21 expression was strongly stimulated by rosiglitazone and genistein in MPH, in agreement with previous studies in hepatocytes or mesenchymal stem cells [ 125 , 126 ]. However, we could not confirm the induction of Fgf21 expression by metformin as previously suggested by other studies [ 127 , 128 ].…”

Obeticholic acid ameliorates obesity and hepatic steatosis by activating brown fat

“…The expression of Zic family member 1 (Zic1), myelin protein zero like 2 (Eva1) or LIM homeobox protein 8 (Lhx8) could not be detected by RT-qPCR in the present study. Therefore, an analysis of the changes between control and treatment groups was not technically possible for these genes in the present study, although previous studies have shown expression of Zic1 in adipogenesis differentiation of C3H10T1/2 cells ( 32 , 33 ). In the future, primary brown adipocyte cells will be used as a model for adipogenesis differentiation in vitro .…”

Bifidobacterium longum subsp. infantis YB0411 Inhibits Adipogenesis in 3T3-L1 Pre-adipocytes and Reduces High-Fat-Diet-Induced Obesity in Mice

“…Differentiated 3T3-L1 cells were prepared in RIPA lysis buffer (containing protease inhibitors), and total protein concentration was estimated using a Protein BCA Assay Kit (Rockford, IL, USA). Western blot analyses were performed as described in our previous study . An anti-β-actin antibody was used as the loading control.…”

“…3T3-L1 cells were transfected with 50 nM AMPKα small interfering RNA (siRNA) (siAMPKα, Santa Cruz Biotechnology) using Lipofectamine 2000 at 80–90% confluency in 6-well plates. The detailed experiments of siAMPKα transfection in 3T3-L1 cells were performed as described elsewhere . The siRNA transfection efficiency was confirmed by qRT-PCR using AMPKα primers (catalog number: sc-29674-PR, Santa Cruz Biotechnology).…”