Objective To observe the effect of S-equol (S-Eq) on sodium oleate (NaOL)-induced steatosis in BRL cells, and to investigate its mechanism of action. Methods BRL cells were cultured in vitro, treated with different concentrations of S-Eq, NaOL and PHTPP for 48h. The appropriate concentration of NaOL for the induction of BRL cell steatosis and the appropriate concentration of S-Eq and PHTPP for intervention were determined by cell viability, oil red O staining and cell TG content. The cells were divided into control group, model group, model + low and high S-Eq concentration (10-6 mol/L, 10-5 mol/L) groups, model + estradiol (E2, 10-7 mol/L) group, model + S-Eq (10-5 mol/L) + Fulvestrant (10-6 mol/L) group. After 48 h of intervention, the contents of TG, TNF-α and IL-6 were detected in cells, the expression levels of TNF-α and IL-6 mRNA were detected by qRT-PCR, and the expression levels of ERβ, PI3k, AKT, p-AKT, p-p-p65 proteins were detected by Western blotting. Results Compared with the control group, when the NaOL concentration was 0.48 mmol/L, the viability of BRL cells did not decrease significantly, the content of TG was significantly increased, and a large number of lipid droplets were accumulated in the cells, which was determined as the appropriate concentration of NaOL for inducing steatosis. At the end of the intervention, compared with the model group, S-Eq or E2 intervention could significantly reduce the contents of TG, TNF-α and IL-6 (P<0.05), as well as the expression levels of TNF-α and IL-6 mRNA in cells (P<0.05). Meanwhile, the expressions of ERβ, PI3k, p-AKT were significant increased, and the expression level of p-p65 protein was significanty reduced (P<0.05). PHTPP could significantly inhibit the improvement effect of S-Eq on cell steatosis. Conclusion S-Eq could effectively improve NAOL-induced BRL cell steatosis, and part of the mechanism is related to the reduction of inflammatory response by S-Eq via regulating the PI3K/AKT/NF-κB signaling pathway through ERβ.
Key words
s-equol /
steatosis /
phosphatidylinositol 3-kinase/serine-threonine protein kinase /
nuclear factor kappa-B
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
References
[1] Bellentani S.The epidemiology of non-alcoholic fatty liver disease[J].Liver Int,2017,37:81–84.
[2] Buzzetti E,Pinzani M,Tsochatzis EA.The multiple-hit pathogenesis of non-alcoholic fatty liver disease (NAFLD)[J]. Metabolism,2016,65:1038–1048.
[3] Della Torre S.Non-alcoholic fatty liver disease as a canonical example of metabolic inflammatory-based liver disease showing a sex-specific prevalence: relevance of estrogen signaling[J]. Front Endocrinol (Lausanne), 2020,11:572490.
[4] Lee C,Kim J,Jung Y.Potential therapeutic application of estrogen in gender disparity of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis[J].Cells,2019,8:1259.
[5] Mayo B,Vazquez L,Florez AB.Equol:a bacterial metabolite from the daidzein isoflavone and its presumed beneficial health effects[J]. Nutrients,2019,11:2231.
[6] 张贵明, 倪向敏, 崔涵强, 等. 雌马酚干预对高脂饮食诱导的去卵巢大鼠非酒精性脂肪肝的影响[J]. 陆军军医大学学报, 2022, 44:2129–2137.
[7] 张新,陈文娜,宋囡,等. 基于分子互作网络探讨丹蒌片干预PI3K/AKT/NF-κB/TNF通路防治非酒精性脂肪肝病的机制[J]. 中国免疫学志,2022,38:1324–1332.
[8] Xu Z, Xu J, Li S, et al. S-Equol enhances osteoblastic bone formation and prevents bone loss through OPG/RANKL via the PI3K/Akt pathway in streptozotocin-induced diabetic rats[J].Front Nutr,2022,9:986192.
[9] Shingina A,DeWitt PE,Dodge JL, et al. Future trends in demand for liver transplant: birth cohort effects among patients with NASH and HCC[J]. Transplantation, 2019,103:140–148.
[10] Pradere JP,Hernandez C,Koppe C, ,et al. Negative regulation of NF-κB p65 activity by serine 536 phos-phorylation[J]. Sci Signal. Negative regulation of NF-κB p65 activity by serine 536 phos-phorylation[J]. Sci Signal, 2016,9:ra85.
[11] Zhang F,Ru N,Shang ZH, et al. Daidzein ameliorates spinal cord ischemia/reperfusion injury-induced neurological function deficits in Sprague-Dawley rats through PI3K/Akt signaling pathway[J]. Exp Ther Med,2017,14:4878–4886.
[12] Huang L,Tang H,Hu J.METTL3 attenuates inflammation in fusarium solani-induced keratitis via the PI3K/AKT signaling pathway[J]. Invest Ophthalmol Vis Sci, 2022,63:20.
[13] Meng Q,Li J,Chao Y, et al. β-estradiol adjusts intestinal function via ERβ and GPR30 mediated PI3K/ AKT signaling activation to alleviate postmenopausal dyslipidemia[J]. Biochem Pharmacol, 2020,180:114134.
[14] Huang LW,Huang TC,Hu YC, et al. S-equol protects chondrocytes against sodium nitroprusside-caused matrix loss and apoptosis through activating PI3K/Akt pathway[J].Int J Mol Sci,2021,22:7054.
[15] Muthyala RS,Ju YH, Sheng S, et al. Equol, a natural estrogenic metabolite from soy isoflavones: convenient preparation and resolution of R- and S-equols and their differing binding and biological activity through estrogen receptors alpha and beta[J]. Bioorg Med Chem, 2004,12:1559–1567.
[16] Weigt C,Hertrampf T,Kluxen FM, et al. Molecular effects of ER alpha- and beta-selective agonists on regulation of energy homeostasis in obese female Wistar rats[J]. Mol Cell Endocrinol,2013,377:147–158.
[17] Qin H,Song Z,Shaukat H, et al. Genistein regulates lipid metabolism via estrogen receptor β and its downstream signal Akt/mTOR in HepG2 cells[J]. Nutrients, 2021,13:4015.
[18] Eyster KM.The estrogen receptors: an overview from different perspectives[J]. Methods Mol Biol, 2016,1366:1–10.
