Salvianolic acid B alleviates oxidative stress in non-alcoholic fatty liver disease by mediating the SIRT3/FOXO1 signaling pathway

doi:10.5246/jcps.2022.09.059

Abstract

Abstract:

Salvianolic acid B (Sal B) is a polyphenolic antioxidant that has been shown to have anti-lipid accumulation, anti-inflammatory, and free oxygen radical scavenging activities in various diseases. Here, we aimed to examine whether Sal B could alleviate non-alcoholic fatty liver disease (NAFLD) and explore the possible mechanisms. Signaling pathways involved in oxidative stress, including SIRT3, SOD2, and FOXO1 pathways, were investigated by Western blotting analysis, RT-qPCR, and immunoprecipitation (IP). In the present study, oleic acid (OA) successfully induced lipid and peroxide accumulation, decreased SIRT3 expression, and increased FOXO1 acetylation. However, Sal B significantly reversed these trends. SIRT3 plasmid transfection further reduced the expression of acetylated FOXO1 and considerably enhanced the regulation of SIRT3 and acetylated FOXO1 induced by Sal B. Following SIRT3 siRNA transfection, Sal B-induced down-regulation of acetylated FOXO1 was blocked, suggesting that Sal B-mediated protection occurred through SIRT3-mediated FOXO1 deacetylation. The SIRT3/FOXO1 pathway was a critical therapeutic target for controlling oxidative stress in NAFLD, and Sal B conferred protection against OA-induced hepatic steatosis and oxidative stress through SIRT3-mediated FOXO1 deacetylation.

Key words: Oleic acid, Oxidative stress, Non-alcoholic fatty liver disease, SIRT3, Acetylated, Salvianolic acid B

Supporting:

Xiaoqing Chen, Bo Peng, Hongmei Jiang, Changxu Zhang, Haiyan Li, Ziyin Li. Salvianolic acid B alleviates oxidative stress in non-alcoholic fatty liver disease by mediating the SIRT3/FOXO1 signaling pathway[J]. Journal of Chinese Pharmaceutical Sciences, 2022, 31(9): 698-710.

Figures/Tables 5

References 42

[1]	Younossi, Z.; Anstee, Q.M.; Marietti, M.; Hardy, T.; Henry, L.; Eslam, M.; George, J.; Bugianesi, E. Global burden of NAFLD and NASH: trends, predictions, risk factors and prevention. Nat. Rev. Gastroenterol. Hepatol. 2018, 15, 11–20.
[2]	Buzzetti, E.; Pinzani, M.; Tsochatzis, E.A. The multiple-hit pathogenesis of non-alcoholic fatty liver disease (NAFLD). Metabolism. 2016, 65, 1038–1048.
[3]	Yan, T.T.; Yan, N.N.; Wang, P.; Xia, Y.L.; Hao, H.P.; Wang, G.J.; Gonzalez, F.J. Herbal drug discovery for the treatment of nonalcoholic fatty liver disease. Acta Pharm. Sin. B. 2020, 10, 3–18.
[4]	Ma, Z.L.; Zhang, W.W.; Fan, W.H.; Wu, Y.R.; Zhang, M.H.; Xu, J.; Li, W.Q.; Sun, L.; Liu, W.J.; Liu, W. Forkhead box O1-mediated ubiquitination suppresses RIG-I-mediated antiviral immune responses. Int. Immunopharmacol. 2021, 90, 107152.
[5]	Murtaza, G.; Khan, A.K.; Rashid, R.; Muneer, S.; Hasan, S.M.F.; Chen, J.X. FOXO transcriptional factors and long-term living. Oxidative Med. Cell Longev. 2017, 2017, 3494289.
[6]	Wang, S.; Xia, P.Y.; Huang, G.L.; Zhu, P.P.; Liu, J.; Ye, B.Q.; Du, Y.; Fan, Z.S. FoxO1-mediated autophagy is required for NK cell development and innate immunity. Nat. Commun. 2016, 7, 11023.
[7]	Rubio, B.; Mora, C.; Pintado, C.; Mazuecos, L.; Fernández, A.; López, V.; Andrés, A.; Gallardo, N. The nutrient sensing pathways FoxO1/3 and mTOR in the heart are coordinately regulated by central leptin through PPARβ/δ. Implications in cardiac remodeling. Metabolism. 2021, 115, 154453.
[8]	Li, P.P.; Song, X.L.; Zhang, D.W.; Guo, N.F.; Wu, C.W.; Chen, K.R.; Liu, Y.; Yuan, L.; Chen, X.L.; Huang, X.Z. Resveratrol improves left ventricular remodeling in chronic kidney disease via Sirt1-mediated regulation of FoxO1 activity and MnSOD expression. BioFactors. 2020, 46, 168–179.
[9]	Xing, Y.Q.; Li, A.; Yang, Y.; Li, X.X.; Zhang, L.N.; Guo, H.C. The regulation of FOXO1 and its role in disease progression. Life Sci. 2018, 193, 124–131.
[10]	Miyauchi, T.; Uchida, Y.; Kadono, K.; Hirao, H.; Kawasoe, J.; Watanabe, T.; Ueda, S.; Okajima, H.; Terajima, H.; Uemoto, S. Up-regulation of FOXO1 and reduced inflammation by β-hydroxybutyric acid are essential diet restriction benefits against liver injury. PNAS. 2019, 116, 13533–13542.
[11]	Dolinsky, V.W. The role of sirtuins in mitochondrial function and doxorubicin-induced cardiac dysfunction. Biol. Chem. 2017, 398, 955–974.
[12]	Xu, X.; Zhu, X.P.; Bai, J.Y.; Xia, P.; Li, Y.; Lu, Y.; Li, X.Y.; Gao, X. Berberine alleviates nonalcoholic fatty liver induced by a high-fat diet in mice by activating SIRT3. FASEB J. 2019, 33, 7289–7300.
[13]	Li, J.Y.; Chen, T.S.; Xiao, M.; Li, N.; Wang, S.J.; Su, H.Y.; Guo, X.B.; Liu, H.; Yan, F.Y.; Yang, Y.; Zhang, Y.; Bu, P.L. Mouse Sirt3 promotes autophagy in AngII-induced myocardial hypertrophy through the deacetylation of FoxO1. Oncotarget. 2016, 7, 86648–86659.
[14]	Hong, M.; Li, S.; Wang, N.; Tan, H.Y.; Cheung, F.; Feng, Y.B. A biomedical investigation of the hepatoprotective effect of radix salviae miltiorrhizae and network pharmacology-based prediction of the active compounds and molecular targets. Int. J. Mol. Sci. 2017, 18, 620.
[15]	Wang, Y.C.; Chen, J.; Kong, W.Z.; Zhu, R.P.; Liang, K.; Kan, Q.X.; Lou, Y.H.; Liu, X.Y. Regulation of SIRT3/FOXO1 signaling pathway in rats with non-alcoholic steatohepatitis by salvianolic acid B. Arch. Med. Res. 2017, 48, 506–512.
[16]	Tariq, Z.; Green, C.J.; Hodson, L. Are oxidative stress mechanisms the common denominator in the progression from hepatic steatosis towards non-alcoholic steatohepatitis (NASH)? Liver Int. 2014, 34, e180–e190.
[17]	Zhang, Z.F.; Chen, H.S.; Li, Z.R. Simultaneous determination of five bioactive phenolic acids in Salvia yunnanensis and Salvia miltiorrhiza by HPLC. J. Chin. Pharm. Sci. 2010, 19, 271–278.
[18]	Li, C.L.; Liu, B.; Wang, Z.Y.; Xie, F.; Qiao, W.; Cheng, J.; Kuang, J.Y.; Wang, Y.; Zhang, M.X.; Liu, D.S. Salvianolic acid B improves myocardial function in diabetic cardiomyopathy by suppressing IGFBP₃. J. Mol. Cell Cardiol. 2020, 139, 98–112.
[19]	Shi, Y.N.; da Pan, Yan, L.H.; Chen, H.Y.; Zhang, X.M.; Yuan, J.H.; Mu, B. Salvianolic acid B improved insulin resistance through suppression of hepatic ER stress in ob/ob mice. Biochem. Biophys. Res. Commun. 2020, 526, 733–737.
[20]	Li, L.; Li, R.; Zhu, R.Y.; Chen, B.B.; Tian, Y.M.; Zhang, H.; Xia, B.K.; Jia, Q.Q.; Wang, L.L.; Zhao, D.D.; Mo, F.F.; Li, Y.; Zhang, S.J.; Gao, S.H.; Zhang, D.W.; Guo, S.Z. Salvianolic acid B prevents body weight gain and regulates gut microbiota and LPS/TLR4 signaling pathway in high-fat diet-induced obese mice. Food Funct. 2020, 11, 8743–8756.
[21]	Wang, L.; Liu, X.L.; Nie, J.; Zhang, J.; Kimball, S.R.; Zhang, H.; Zhang, W.J.; Jefferson, L.S.; Cheng, Z.N.; Ji, Q.H.; Shi, Y.G. ALCAT1 controls mitochondrial etiology of fatty liver diseases, linking defective mitophagy to steatosis. Hepatology. 2015, 61, 486–496.
[22]	Jha, S.K.; Jha, N.K.; Kumar, D.; Ambasta, R.K.; Kumar, P. Linking mitochondrial dysfunction, metabolic syndrome and stress signaling in Neurodegeneration. Biochim. Biophys. Acta BBA Mol. Basis Dis. 2017, 1863, 1132–1146.
[23]	Ajith, T.A. Role of mitochondria and mitochondria-targeted agents in non-alcoholic fatty liver disease. Clin. Exp. Pharmacol. Physiol. 2018, 45, 413–421.
[24]	Wang, P.J.; Xu, S.Y.; Li, W.Z.; Wang, F.; Yang, Z.; Jiang, L.C.; Wang, Q.L.; Huang, M.Q.; Zhou, P. Salvianolic acid B inhibited PPARγ expression and attenuated weight gain in mice with high-fat diet-induced obesity. Cell Physiol. Biochem. 2014, 34, 288–298.
[25]	Wang, R.; Yu, X.Y.; Guo, Z.Y.; Wang, Y.J.; Wu, Y.; Yuan, Y.F. Inhibitory effects of salvianolic acid B on CCl₄-induced hepatic fibrosis through regulating NF-κB/IκBα signaling. J. Ethnopharmacol. 2012, 144, 592–598.
[26]	Zhai, J.H.; Tao, L.N.; Zhang, Y.M.; Gao, H.; Qu, X.Y.; Song, Y.Q.; Zhang, S.X. Salvianolic acid B attenuates apoptosis of HUVEC cells treated with high glucose or high fat via Sirt1 activation. Evid. Based Complementary Altern. Med. 2019, 2019, 9846325.
[27]	Ma, Z.G.; Xia, H.Q.; Cui, S.L.; Yu, J. Attenuation of renal ischemic reperfusion injury by salvianolic acid B via suppressing oxidative stress and inflammation through PI3K/Akt signaling pathway. Braz. J. Med. Biol. Res. 2017, 50, e5954.
[28]	Zhang, X.S.; Wu, Q.; Lu, Y.; Wan, J.R.; Dai, H.B.; Zhou, X.M.; Lv, S.Y.; Chen, X.M.; Zhang, X.; Hang, C.H.; Wang, J. Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways. Free. Radic. Biol. Med. 2018, 124, 504–516.
[29]	Li, J.; Li, X.F.; Li, Z.K.; Zhang, L.; Liu, Y.G.; Ding, H.; Yin, S.Y. Isofraxidin, a coumarin component improves high-fat diet induced hepatic lipid homeostasis disorder and macrophage inflammation in mice. Food Funct. 2017, 8, 2886–2896.
[30]	Yoo, A.; Narayan, V.P.; Hong, E.Y.; Whang, W.K.; Park, T. Scopolin ameliorates high-fat diet induced hepatic steatosis in mice: potential involvement of SIRT1-mediated signaling cascades in the liver. Sci. Rep. 2017, 7, 2251.
[31]	Nassir, F.; Arndt, J.J.; Johnson, S.A.; Ibdah, J.A. Regulation of mitochondrial trifunctional protein modulates nonalcoholic fatty liver disease in mice. J. Lipid Res. 2018, 59, 967–973.
[32]	Kwon, D.N.; Park, W.J.; Choi, Y.J.; Gurunathan, S.; Kim, J.H. Oxidative stress and ROS metabolism via down-regulation of sirtuin 3 expression in Cmah-null mice affect hearing loss. Aging. 2015, 7, 579–594.
[33]	Ommer, J.; Selfe, J.L.; Wachtel, M.; O'Brien, E.M.; Laubscher, D.; Roemmele, M.; Kasper, S.; Delattre, O.; Surdez, D.; Petts, G.; Kelsey, A.; Shipley, J.; Schäfer, B.W. Aurora A kinase inhibition destabilizes PAX3-FOXO1 and MYCN and synergizes with navitoclax to induce rhabdomyosarcoma cell death. Cancer Res. 2020, 80, 832–842.
[34]	Elkhwanky, M.S.; Hakkola, J. Extranuclear sirtuins and metabolic stress. Antioxid. Redox Signal. 2018, 28, 662–676.
[35]	Zhang, L.Q.; Zhang, Z.G.; Li, C.B.; Zhu, T.T.; Gao, J.; Zhou, H.; Zheng, Y.Z.; Chang, Q.; Wang, M.S.; Wu, J.Y.; Ran, L.Y.; Wu, Y.J.; Miao, H.L.; Zou, X.J.; Liang, B. S100A11 promotes liver steatosis via FOXO1-mediated autophagy and lipogenesis. Cell Mol. Gastroenterol. Hepatol. 2021, 11, 697–724.
[36]	Kwon, D.N.; Park, W.J.; Choi, Y.J.; Gurunathan, S.; Kim, J.H. Oxidative stress and ROS metabolism via down-regulation of sirtuin 3 expression in Cmah-null mice affect hearing loss. Aging. 2015, 7, 579–594.
[37]	Tiwari, H.S.; Misra, U.K.; Kalita, J.; Mishra, A.; Shukla, S. Oxidative stress and glutamate excitotoxicity contribute to apoptosis in cerebral venous sinus thrombosis. Neurochem. Int. 2016, 100, 91–96.
[38]	Chang, Y.W.; Zhao, Y.F.; Cao, Y.L.; Gu, X.F.; Li, Z.Q.; Wang, S.Q.; Miao, J.H.; Zhan, H.S. Liver X receptor a inhibits osteosarcoma cell proliferation through up-regulation of FoxO1. Cell Physiol. Biochem. 2013, 32, 180–186.
[39]	Matsuzaki, H.; Lee, S.N.; Maeda, M.; Kumagai-Takei, N.; Nishimura, Y.; Otsuki, T. FoxO1 regulates apoptosis induced by asbestos in the MT-2 human T-cell line. J. Immunotoxicol. 2016, 13, 620–627.
[40]	Liu, L.H.; Zheng, L.D.; Zou, P.; Brooke, J.; Smith, C.; Long, Y.C.; Almeida, F.A.; Liu, D.M.; Cheng, Z.Y. FoxO1 antagonist suppresses autophagy and lipid droplet growth in adipocytes. Cell Cycle Georget. Tex. 2016, 15, 2033–2041.
[41]	Li, J.Y.; Chen, T.S.; Xiao, M.; Li, N.; Wang, S.J.; Su, H.Y.; Guo, X.B.; Liu, H.; Yan, F.Y.; Yang, Y.; Zhang, Y.; Bu, P.L. Mouse Sirt3 promotes autophagy in AngII-induced myocardial hypertrophy through the deacetylation of FoxO1. Oncotarget. 2016, 7, 86648–86659.
[42]	Koltai, E.; Bori, Z.; Osvath, P.; Ihasz, F.; Peter, S.; Toth, G.; Degens, H.; Rittweger, J.; Boldogh, I.; Radak, Z. Master athletes have higher miR-7, SIRT3 and SOD2 expression in skeletal muscle than age-matched sedentary controls. Redox Biol. 2018, 19, 46–51.