Modified SJH alleviates FFAs-induced hepatic steatosis through leptin signaling pathways

Abstract

Samjunghwan (SJH) is an herbal formula used in traditional Korean medicine. This prescription has long been used in treatment of aging and lifestyle diseases. The current study showed the effect and mechanisms of anti-hepatic steatosis action of modified SJH (mSJH) in vitro and in vivo. Treatment with mSJH resulted in significantly decreased intracellular lipid accumulation in steatosis-induced cells. Furthermore, mSJH triggered the phosphorylation of AMP-activated protein kinase and acetyl-CoA carboxylase as well as increased the expression of leptin at both protein and gene levels. In addition, C57BL6 mice fed high-fat diet (HFD) showed significant improvements in body, liver weights and fat weights; and serum, hepatic and fecal lipid parameters in response to the treatment with mSJH. Furthermore, mSJH showed favorable effects on the hepatic expression of several genes related to lipid metabolism. Betaine, one of constituents of mSJH exerted fundamental beneficial impact on FFAs-induced cells. However, the beneficial effects of mSJH were diminished upon blocking of leptin signaling by dexamethasone, suggesting the leptin signaling as a key component in mSJH-mediated modulation of lipid homeostasis. Our results suggest that mSJH exerts an anti-hepatic steatosis effect via activation of leptin and associated signaling cascades related to lipid metabolism.

A Controlled Fermented Samjunghwan Herbal Formula Ameliorates Non-alcoholic Hepatosteatosis in HepG2 Cells and OLETF Rats

“…The DPPH assay was performed to determine the free radical scavenging activity of SJH, FSJH, or PMM of bacterial strains, according to our previous report ( Lim et al, 2017 ). Briefly, 40 μl of SJH or FSJH extracts or PMM of bacterial strains were added to 760 μl ethanolic DPPH solution (0.3 mM), while an equal amount of ethanol without sample and DPPH served as control.…”

“…For cell viability analysis, HepG2 cells were seeded at a density of 5 × 10 4 cells (100 μL) in 96-well plates, cultured for 24 h. HepG2 cells were treated with extracts of SJH or FSJH (50, 100, 200, or 400 μg/ml) or PMM (1% or 2% v/v) for 24 h under the same culturing conditions described above. Following this, cell culture media was replaced with fresh media, after which the cell viability was measured using an Ez-cytox assay kit according to manufacturer’s instructions and our previous report ( Lim et al, 2017 ). For other experimental purposes, HepG2 cells were seeded at a density of 5 × 10 5 cells (1 mL) in 12-well plates, and then cultured for 24 h. The cell culture media was then replaced with fresh media, after which cells were induced with a mixture of FFAs (1 mM/ml; OA:PA = 2:1 ratio) and treated with extracts of SJH or FSJH (400 μg/ml), or PMM of bacterial strains (1% v/v) for 24 h under the same culturing conditions described above and a series of experimental analyses were performed as described below.…”

“…After treatment with extracts of SJH or FSJH, or PMM of bacterial strains for 24 h, HepG2 cells were carefully washed twice with cold DPBS without disturbing the cells and subsequently fixed with 10% formalin solution for 5 min at room temperature. The cells were then washed gently with 60% isopropanol, after which they were stained with the working solution of oil red o in 60% isopropanol for 15 min ( Lim et al, 2017 ). Next, stained cells were washed several times with cold distilled water and finally examined under an inverted microscope (DMI 6000, Leica, Jena, Germany) and images were acquired.…”

“…Mori fructus , the major constituent of SJH, has been shown to protect against obesity and related complications ( Peng et al, 2011 ; Jiao et al, 2014 ), and the hypolipidemic effects of this herb are thought to be mediated through modulation of lipid homeostasis ( Liu et al, 2009 ). We recently found that modified SJH alleviated FFAs-induced HS and that this effect was mediated through leptin signaling pathways ( Lim et al, 2017 ).…”

Lactate-Fortified Puerariae Radix Fermented by Bifidobacterium breve Improved Diet-Induced Metabolic Dysregulation via Alteration of Gut Microbial Communities

“…We also observed a significant reduction in the hepatic expression of PPARγ gene in the HFD+F group upon exposure to XEN and fPR, but not to PR. In an earlier study, using HepG2 cells as a model, we showed that down-regulation of PPARγ may contribute to the protective effect of herbal medicines against free fatty acids-induced hepatic steatosis [68]. On the other hand, AMPK, the function of which has been extensively studied in muscles and liver, has a central role in maintaining cellular energy homeostasis [8,39,69].…”