Yu Gan Long Ameliorates Hepatic Fibrosis by Inhibiting PI3K/AKT, Ras/ERK and JAK1/STAT3 Signaling Pathways in CCl₄-induced Liver Fibrosis Rats

Abstract

Yu Gan Long (YGL) is a Chinese traditional herbal formula which has been reported to attenuate liver fibrosis for many years and we have explored its anti-fibrotic mechanism through blocking transforming growth factor (TGF-β) in the previous study. But the mechanisms associated with platelet-derived growth factor (PDGF)-BB remain obscure. In this study, we further investigated the mechanism of YGL reducing carbon tetrachloride (CCl₄)-induced liver fibrosis in rats. Our results showed that YGL suppressed CCl₄-induced upregulation of collagen IV (Col IV), type HI precollagen (PCHI), hyaluronuc acid (HA) and laminin (LN), which are implicated in liver fibrosis. Also, YGL reduced the α-smooth muscle actin (α-SMA) expression, which acts as the indicator of liver fibrosis. Furthermore, YGL decreased the serum levels of hepatic stellate cell (HSC) mitogen PDGF-BB and inflammation cytokines, including TNF-α, IL-1β, IL-6. Markers involved in liver fibrosis, such as Ras, p-Raf-1, p-ERK1/2, p-JNK, p-P38, p-PI3K, p-AKT, p-JAKl, p-STAT3 were downregulated significantly after treatment with YGL. Our results indicated that YGL ameliorated CCl₄-induced liver fibrosis by reducing inflammation cytokines production, and suppressing Ras/ERK, PI3K/AKT, and JAK1/STAT3 signaling pathways, which provided further evidence towards elucidation of the anti-fibrotic mechanism of YGL.

Keywords

Yu Gan Long / traditional Chinese herbal therapy / liver fibrosis / Ras/ERK / PI3K/AKT / JAK1/STAT3

References

[1] RockeyD, FriedmanS. Hepatic Fibrosis and Cirrhosis. Dig Dis, 2006, 33(33): 87-109

[2] BatallerR, BrennerDA. Liver fibrosis. J Clin Invest, 2005, 115(2): 209-218

[3] LiuJY, ChenCC, WangWH, et al.. The protective effects of Hibiscus sabdariffa extract on CCl₄-induced liver fibrosis in rats. Food Chem Toxicol, 2006, 44(3): 336-343

[4] WangQL, YuanLJ, TaoYY, et al.. Fuzheng Huayu recipe and vitamin E reverse renal interstitial fibrosis through counteracting TGF-β1-induced epithelial-to-mesenchymal transition. J Ethnopharmacol, 2010, 127(3): 631-640

[5] LiuC, HuY, XuL, et al.. Effect of Fuzheng Huayu formula and its actions against liver fibrosis. Chin Med, 2009, 4: 12

[6] Wang DM, Chang FR, Shen MQ, et al. Treatment of 60 Cases of Hepatitis B Virus Carrier with “Yuganlong” and Its Experimental Study. Jiangsu J Tradit Chin Med (Chinese), 1999(08):45–46

[7] Cui XY, Zhang LJ. The clinical experience of treating 50 cases of hepatitis b with yuganlong oral liquid. Qinghai Med J (Chinese), 1998(12):44–45

[8] XiaY, YuB, MaC, et al.. Yu Gan Long reduces rat liver fibrosis by blocking TGF-β1/Smad pathway and modulating the immunity. Biomed Pharmacother, 2018, 106: 1332-1338

[9] DirksRP, BloemersHP. Signals controlling the expression of PDGF. Mol Biol Rep, 1995, 22(1): 1-24

[10] ZhouJ, DengY, YanL, et al.. Serum platelet-derived growth factor BB levels: A potential biomarker for the assessment of liver fibrosis in patients with chronic hepatitis B. Int J Infect Dis, 2016, 49: 94-99

[11] WangY, GaoJ, ZhangD, et al.. New insights into the antifibrotic effects of sorafenib on hepatic stellate cells and liver fibrosis. J Hepatol, 2010, 53(1): 132-144

[12] Borkham-KamphorstE, HerrmannJ, StollD, et al.. Dominant-negative soluble PDGF-receptor inhibits hepatic stellate cell activation and attenuates liver fibrosis. Lab Invest, 2004, 84(6): 766-777

[13] PengR, YuanY. Antifibrotic effects of tanshinol in experimental hepatic fibrosis by targeting PI3K/AKT/mTOR/p70S6K1 signaling pathways. Discov Med, 2017, 23(125): 81-94

[14] WangX, IkejimaK, KonK, et al.. Ursolic acid ameliorates hepatic fibrosis in the rat by specific induction of apoptosis in hepatic stellate cells. J Hepatol, 2010, 55(2): 379-387

[15] CalvisiD, LaduS, GordenA, et al.. Ubiquitous Activation of Ras and Jak/Stat Pathways in Human HCC. Gastroenterology, 2006, 130(4): 1117-1128

[16] YuZ, TangL, ChenL, et al.. Capillarisin Suppresses Lipopolysaccharide-Induced Inflammatory Mediators in BV2 Microglial Cells by Suppressing TLR4-Mediated NF-κB and MAPKs Signaling Pathway. Neurochem Res, 2015, 40(6): 1095-1101

[17] LuJ, ZhongY, LinZ, et al.. Baicalin alleviates radiation-induced epithelial-mesenchymal transition of primary type II alveolar epithelial cells via TGF-β and ERK/GSK3β signaling pathways. Biomed Pharmacother, 2017, 95: 1219-1224

[18] HuangX, HeY, ChenY, et al.. Baicalin attenuates bleomycin-induced pulmonary fibrosis via adenosine A2a receptor related TGF-β1-induced ERK1/2 signaling pathway. BMC Pulm Med, 2016, 16(1): 132

[19] MaC, ZhuL, WangJ, et al.. Anti-inflammatory effects of water extract of Taraxacum mongolicum hand.-Mazz on lipopolysaccharide-induced inflammation in acute lung injury by suppressing PI3K/Akt/mTOR signaling pathway. J Ethnopharmacol, 2015, 168: 349-355

[20] MaC, ZouL, XiaY, et al.. Extracts of Coleus forskohlii relieves cough and asthma symptoms via modulating inflammation and the extracellular matrix. J Cell Biochem, 2019, 120(6): 9648-9655

[21] SmythR, Munday MichaelR, York MalcolmJ, et al.. Comprehensive characterization of serum clinical chemistry parameters and the identification of urinary superoxide dismutase in a carbon tetrachloride-induced model of hepatic fibrosis in the female Hanover Wistar rat. Int J Exp Pathol, 2007, 88(5): 361-376

[22] JayakumarT, RameshE, GeraldineP. Antioxidant activity of the oyster mushroom, Pleurotus ostreatus, on CCl₄-induced liver injury in rats. Food Chem Toxicol, 2006, 44(12): 1989-1996

[23] CarpinoG, MoriniS, GinannicorradiniS, et al.. Alpha-SMA expression in hepatic stellate cells and quantitative analysis of hepatic fibrosis in cirrhosis and in recurrent chronic hepatitis after liver transplantation. Dig Liver Dis, 2005, 37(5): 349-356

[24] McCubreyJA, SteelmanLS, ChappellWH, et al.. Roles of the Raf/MEK/ERK pathway in cell growth, malignant transformation and drug resistance. Biochim Biophys Acta, 2007, 1773(8): 1263-1284

[25] PoliG. Pathogenesis of liver fibrosis: role of oxidative stress. Mol Aspects Med, 2000, 21(3): 49-98

[26] HuB, WangSS, DuQ. Traditional Chinese medicine for prevention and treatment of hepatocarcinoma: From bench to bedside. World J Hepatol, 2015, 7(9): 1209-1232

[27] Borkham-KamphorstE, WeiskirchenR. The PDGF system and its antagonists in liver fibrosis. Cytokine Growth Factor Rev, 2016, 28: 53-61

[28] YoshidaS, IkenagaN, LiuSB, et al.. Extrahepatic platelet-derived growth factor-beta, delivered by platelets, promotes activation of hepatic stellate cells and biliary fibrosis in mice. Gastroenterology, 2014, 147(6): 1378-1392

[29] CampoJAD, GallegoP, GrandeL. Role of inflammatory response in liver diseases: Therapeutic strategies. World J Hepatol, 2018, 10(1): 1-7

[30] YinC, EvasonKJ, AsahinaK, et al.. Hepatic stellate cells in liver development, regeneration, and cancer. J Clin Invest, 2013, 123(5): 1902-1910

[31] AhnJ, ChaeHS, ChinYW, et al.. Furylhydroquinones and miscellaneous compounds from the roots of Lithospermum erythrorhizon and their anti-inflammatory effect in HaCaT cells. Nat Prod Res, 2018, 31: 1-8

[32] ParkEJ, ChunJN, KimSH, et al.. Schisandrin B suppresses TGFβ1 signaling by inhibiting Smad2/3 and MAPK pathways. Biochem Pharmacol, 2012, 83(3): 378-384

[33] KidgerAM, SipthorpJ, CookSJ. ERK1/2 inhibitors: New weapons to inhibit the RAS-regulated RAF-MEK1/2-ERK1/2 pathway. Pharmacol Ther, 2018, 187: 45-60

[34] FooNP, LinSH, LeeYH, et al.. α-Lipoic acid inhibits liver fibrosis through the attenuation of ROS-triggered signaling in hepatic stellate cells activated by PDGF and TGF-β. Toxicology, 2011, 282(1–2): 39-46

[35] SonG, HinesI, LindquistJ, et al.. Inhibition of Phosphatidylinositol 3-Kinase Signaling in Hepatic Stellate Cells Blocks the Progression of Hepatic Fibrosis. Hepatology, 2009, 50(5): 1512-1523

[36] PiguetAC, DufourJF. PI(3)K/PTEN/AKT pathway. J Hepatol, 2011, 54(6): 1317-1319