Ferulic acid maintains the self-renewal capacity of embryo stem cells and adipose-derived mesenchymal stem cells in high fat diet-induced obese mice

Introduction

A mechanistic study of self-renewal and differentiation provides an understanding of ESCs, which can then be applied as a therapeutic strategy for ESCs
Leukemia inhibitory factor (LIF) is a key regulator in the differentiation and proliferation of both ESCs and somatic stem cells (SSCs)
Nanog is a homeodomain gene that maintains and regulates the development of both cells
Ferulic acid reduces the expression of IL-1β and TNF-α, as well as other known inflammatory genes in LPS activated monocyte-derived macrophages
Reduced the effects of obesity, insulin resistance, hyperlipidemia, hyperglycemia
Reduces rat metabolic syndrome in HFD and high-fructose diet-induced obesity
Obesity is observed as the abnormal distribution and massive accumulation of fat tissues

Mouse ESCs of E14 were cultured in Dulbecco's modified Eagle’s medium (DMEM) supplemented with 15% fetal bovine serum, 2% penicillin-streptomycin, 1% nucleoside mix, L-glutamine, 10−4 M of β-mercaptoethanol, and 103 U/ml of LIF.
All cells were incubated in an atmosphere containing 5% CO2 at 37°C, and natural products (Ferulic acid, xanthohumol, curcumin, ascorbic acid, and quercetin) were treated in a mouse ESCs culture system, followed by staining using an AP detection kit.

Mouse ADMSCs isolation was performed under a laminar flow hood using sterile techniques
Briefly, epididymal fat pads were dissected from each individual mouse, minced extensively in PBS, and subjected to enzymatic digestion in 0.2% Type I Collagenase for 45 min with repeated trituration.
After adding the inhibition solution (20% FBS in HBSS), the samples were spun for 5 min at 450 g and filtered through a strainer, 70 μm in size.

After fasting the mice for 16-18 h, a glucose tolerance test (GTT) was performed using an intraperitoneal (IP) injection (1.5 g/kg body weight) of glucose (Sigma-Aldrich, St. Louis, Missouri, USA).
Mouse blood samples were collected through a cut at the tip of the tail before and at 15, 30, 45, 60, and 120 min after glucose IP injection.

The screening data of AP+ staining with five natural compounds were analyzed by one-way analysis of variance (ANOVA) using Prims software (version 5, USA). The results are presented as the mean ± SE.
For data analysis in an animal experiment, the results are expressed as the means ± SD for 10 mice per group.

Five types of natural compounds (ferulic acid, xanthohumol, curcumin, ascorbic acid, and quercetin) were screened using an alkaline phosphatase (AP+) assay in a mouse ESCs culture system
Ferulic acid was chosen as the top-hit of screening
Among the compounds tested, ferulic acid induced the highest AP+ staining rate for the self-renewal of mouse ESC’s
To confirm this ability, the mice ESCs were cultured with or without LIF, and examined by AP + staining alone
The data showed that ferilic acid affects the expression of Nanog mRNA of mouse ESs, following LIF withdrawal

These results show that ferulic acid maintains self-renewal in mouse ESCs.
It upregulated Nanog mRNA expression and increased FGF2, FN1, and TGF mRNA expression in human ADMSCs, which suggest that it regulates the stem cell ability by not only NANOG, but also other related genes (e.g., TGFβ-1).
The combination of Ferulic acid with a HFD maintained the ADMSC populations in mice compared to HFD alone, suggesting that it may be a natural compound for a therapeutic approach with anti-obesity effects.

Reference

Summarized Research Articles