Estrogen receptor‑β‑dependent effects of saikosaponin‑d on the suppression of oxidative stress‑induced rat hepatic stellate cell activation

Que,Renye; Shen,Yanting; Ren,Jianlin; Tao,Zhihui; Zhu,Xiaoyan; Li,Yong

doi:10.3892/ijmm.2017.3349

Estrogen receptor‑β‑dependent effects of saikosaponin‑d on the suppression of oxidative stress‑induced rat hepatic stellate cell activation

Authors:
- Renye Que
- Yanting Shen
- Jianlin Ren
- Zhihui Tao
- Xiaoyan Zhu
- Yong Li
View Affiliations

Affiliations: Department of Gastroenterology, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200071, P.R. China, Department of Scientific Research, Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200071, P.R. China, Department of Physiology, The Second Military Medical University, Shanghai 200433, P.R. China
Published online on: December 22, 2017 https://doi.org/10.3892/ijmm.2017.3349
Pages: 1357-1364
Copyright: © Que et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Saikosaponin-d (SSd) is one of the major triterpenoid saponins derived from Bupleurum falcatum L., which has been reported to possess antifibrotic activity. At present, there is little information regarding the potential target of SSd in hepatic stellate cells (HSCs), which serve an important role in excessive extracellular matrix (ECM) deposition during the pathogenesis of hepatic fibrosis. Our recent study indicated that SSd may be considered a novel type of phytoestrogen with estrogen‑like actions. Therefore, the present study aimed to investigate the effects of SSd on the proliferation and activation of HSCs, and the underlying mechanisms associated with estrogen receptors. In the present study, a rat HSC line (HSC‑T6) was used and cultured with dimethyl sulfoxide, SSd, or estradiol (E2; positive control), in the presence or absence of three estrogen receptor (ER) antagonists [ICI‑182780, methylpiperidinopyrazole (MPP) or (R,R)-tetrahydrochrysene (THC)], for 24 h as pretreatment. Oxidative stress was induced by exposure to hydrogen peroxide for 4 h. Cell proliferation was assessed by MTT growth assay. Malondialdehyde (MDA), CuZn-superoxide dismutase (CuZn-SOD), tissue inhibitor of metalloproteinases-1 (TIMP-1), matrix metalloproteinase-1 (MMP-1), transforming growth factor-β1 (TGF-β1), hydroxyproline (Hyp) and collagen-1 (COL1) levels in cell culture supernatants were determined by ELISA. Reactive oxygen species (ROS) was detected by ﬂow cytometry. Total and phosphorylated mitogen-activated protein kinases (MAPKs) and α-smooth muscle actin (α-SMA) were examined by western blot analysis. TGF-β1 mRNA expression was determined by RT-quantitative (q)PCR. SSd and E2 were able to significantly suppress oxidative stress‑induced proliferation and activation of HSC‑T6 cells. Furthermore, SSd and E2 were able to reduce ECM deposition, as demonstrated by the decrease in transforming growth factor‑β1, hydroxyproline, collagen‑1 and tissue inhibitor of metalloproteinases‑1, and by the increase in matrix metalloproteinase‑1. These results suggested that the possible molecular mechanism could involve downregulation of the reactive oxygen species/mitogen‑activated protein kinases signaling pathway. Finally, the effects of SSd and E2 could be blocked by co‑incubation with ICI‑182780 or THC, but not MPP, thus indicating that ERβ may be the potential target of SSd in HSC‑T6 cells. In conclusion, these findings suggested that SSd may suppress oxidative stress‑induced activation of HSCs, which relied on modulation of ERβ.

View Figures

View References

1	Bataller R and Brenner DA: Liver fibrosis. J Clin Invest. 115:209–218. 2005. View Article : Google Scholar : PubMed/NCBI
2	Sánchez-Valle V, Chávez-Tapia NC, Uribe M and Méndez-Sánchez N: Role of oxidative stress and molecular changes in liver fibrosis: A review. Curr Med Chem. 19:4850–4860. 2012. View Article : Google Scholar : PubMed/NCBI
3	Li JT, Liao ZX, Ping J, Xu D and Wang H: Molecular mechanism of hepatic stellate cell activation and antifibrotic therapeutic strategies. J Gastroenterol. 43:419–428. 2008. View Article : Google Scholar : PubMed/NCBI
4	Schon HT, Bartneck M, Borkham-Kamphorst E, Nattermann J, Lammers T, Tacke F and Weiskirchen R: Pharmacological Intervention in Hepatic Stellate Cell Activation and Hepatic Fibrosis. Front Pharmacol. 7:332016. View Article : Google Scholar : PubMed/NCBI
5	Ghatak S, Biswas A, Dhali GK, Chowdhury A, Boyer JL and Santra A: Oxidative stress and hepatic stellate cell activation are key events in arsenic induced liver fibrosis in mice. Toxicol Appl Pharmacol. 251:59–69. 2011. View Article : Google Scholar
6	Mormone E, George J and Nieto N: Molecular pathogenesis of hepatic fibrosis and current therapeutic6 approaches. Chem Biol Interact. 193:225–231. 2011. View Article : Google Scholar : PubMed/NCBI
7	Yasuda M, Shimizu I, Shiba M and Ito S: Suppressive effects of estradiol on dimethylnitrosamine-induced fibrosis of the liver in rats. Hepatology. 29:719–727. 1999. View Article : Google Scholar : PubMed/NCBI
8	Xu JW, Gong J, Chang XM, Luo JY, Dong L, Hao ZM, Jia A and Xu GP: Estrogen reduces CCl₄-induced liver fibrosis in rats. World J Gastroenterol. 8:883–887. 2002. View Article : Google Scholar : PubMed/NCBI
9	Shimizu I, Mizobuchi Y, Yasuda M, Shiba M, Ma YR, Horie T, Liu F and Ito S: Inhibitory effect of oestradiol on activation of rat hepatic stellate cells in vivo and in vitro. Gut. 44:127–136. 1999. View Article : Google Scholar
10	Itagaki T, Shimizu I, Cheng X, Yuan Y, Oshio A, Tamaki K, Fukuno H, Honda H, Okamura Y and Ito S: Opposing effects of oestradiol and progesterone on intracellular pathways and activation processes in the oxidative stress induced activation of cultured rat hepatic stellate cells. Gut. 54:1782–1789. 2005. View Article : Google Scholar : PubMed/NCBI
11	Cuzick J: Hormone replacement therapy and the risk of breast cancer. Eur J Cancer. 44:2344–2349. 2008. View Article : Google Scholar : PubMed/NCBI
12	Barnes PJ: Corticosteroids: The drugs to beat. Eur J Pharmacol. 533:2–14. 2006. View Article : Google Scholar : PubMed/NCBI
13	Lee JK, Kim JH and Shin HK: Therapeutic effects of the oriental herbal medicine Sho-saiko-to on liver cirrhosis and carcinoma. Hepatol Res. 41:825–837. 2011. View Article : Google Scholar : PubMed/NCBI
14	Deng G, Kurtz RC, Vickers A, Lau N, Yeung KS, Shia J and Cassileth B: A single arm phase II study of a Far-Eastern traditional herbal formulation (sho-sai-ko-to or xiao-chai-hu-tang) in chronic hepatitis C patients. J Ethnopharmacol. 136:83–87. 2011. View Article : Google Scholar : PubMed/NCBI
15	Qin F, Liu JY and Yuan JH: Chaihu-Shugan-San, an oriental herbal preparation, for the treatment of chronic gastritis: A meta-analysis of randomized controlled trials. J Ethnopharmacol. 146:433–439. 2013. View Article : Google Scholar : PubMed/NCBI
16	Fan J, Li X, Li P, Li N, Wang T, Shen H, Siow Y, Choy P and Gong Y: Saikosaponin-d attenuates the development of liver fibrosis by preventing hepatocyte injury. Biochem Cell Biol. 85:189–195. 2007. View Article : Google Scholar : PubMed/NCBI
17	Dang SS, Wang BF, Cheng YA, Song P, Liu ZG and Li ZF: Inhibitory effects of saikosaponin-d on CCl₄-induced hepatic fibrogenesis in rats. World J Gastroenterol. 13:557–563. 2007. View Article : Google Scholar : PubMed/NCBI
18	Chen MF, Huang CC, Liu PS, Chen CH and Shiu LY: Saikosaponin a and saikosaponin d inhibit proliferation and migratory activity of rat HSC-T6 cells. J Med Food. 16:793–800. 2013. View Article : Google Scholar : PubMed/NCBI
19	Wang P, Ren J, Tang J, Zhang D, Li B and Li Y: Estrogen-like activities of saikosaponin-d in vitro: A pilot study. Eur J Pharmacol. 626:159–165. 2010. View Article : Google Scholar
20	Richter DU, Mylonas I, Toth B, Scholz C, Briese V, Friese K and Jeschke U: Effects of phytoestrogens genistein and daidzein on progesterone and estrogen (estradiol) production of human term trophoblast cells in vitro. Gynecol Endocrinol. 25:32–38. 2009. View Article : Google Scholar : PubMed/NCBI
21	Gille JJ and Joenje H: Cell culture models for oxidative stress: Superoxide and hydrogen peroxide versus normobaric hyperoxia. Mutat Res. 275:405–414. 1992. View Article : Google Scholar : PubMed/NCBI
22	Kiyoshima T, Enoki N, Kobayashi I, Sakai T, Nagata K, Wada H, Fujiwara H, Ookuma Y and Sakai H: Oxidative stress caused by a low concentration of hydrogen peroxide induces senescence-like changes in mouse gingival fibroblasts. Int J Mol Med. 30:1007–1012. 2012. View Article : Google Scholar : PubMed/NCBI
23	Coates D, Zafar S and Milne T: Quantitative real-time gene profiling of human alveolar osteoblasts. Methods Mol Biol. 1537:447–459. 2017. View Article : Google Scholar
24	Lechuga CG, Hernández-Nazara ZH, Domínguez Rosales JA, Morris ER, Rincón AR, Rivas-Estilla AM, Esteban-Gamboa A and Rojkind M: TGF-beta1 modulates matrix metalloproteinase-13 expression in hepatic stellate cells by complex mechanisms involving p38MAPK, PI3-kinase, AKT, and p70S6k. Am J Physiol Gastrointest Liver Physiol. 287:G974–G987. 2004. View Article : Google Scholar : PubMed/NCBI
25	Iredale JP: Hepatic stellate cell behavior during resolution of liver injury. Semin Liver Dis. 21:427–436. 2001. View Article : Google Scholar : PubMed/NCBI
26	Zhang BZ, Guo XT, Chen JW, Zhao Y, Cong X, Jiang ZL, Cao RF, Cui K, Gao SS and Tian WR: Saikosaponin-D attenuates heat stress-induced oxidative damage in LLC-PK1 cells by increasing the expression of anti-oxidant enzymes and HSP72. Am J Chin Med. 42:1261–1277. 2014. View Article : Google Scholar : PubMed/NCBI
27	Zhao L, Zhang H, Bao J, Liu J and Ji Z: Saikosaponin-d protects renal tubular epithelial cell against high glucose induced injury through modulation of SIRT3. Int J Clin Exp Med. 8:6472–6481. 2015.PubMed/NCBI
28	Hong IH, Park SJ, Goo MJ, Lee HR, Park JK, Ki MR, Kim SH, Lee EM, Kim AY and Jeong KS: JNK1 and JNK2 regulate α-SMA in hepatic stellate cells during CCl₄-induced fibrosis in the rat liver. Pathol Int. 63:483–491. 2013. View Article : Google Scholar : PubMed/NCBI
29	Varela-Rey M, Montiel-Duarte C, Osés-Prieto JA, López-Zabalza MJ, Jaffrèzou JP, Rojkind M and Iraburu MJ: p38 MAPK mediates the regulation of alpha1(I) procollagen mRNA levels by TNF-alpha and TGF-beta in a cell line of rat hepatic stellate cells(1). FEBS Lett. 528:133–138. 2002. View Article : Google Scholar : PubMed/NCBI
30	Friedman SL: Liver fibrosis - from bench to bedside. J Hepatol. 38(Suppl 1): S38–S53. 2003. View Article : Google Scholar
31	Bhaskar ME: Management of cirrhosis and ascites. N Engl J Med. 351:300–301; author reply 300-301, 2004. PubMed/NCBI
32	O'Beirne JP, Foxton MR and Heneghan MA: Management of cirrhosis and ascites. N Engl J Med. 351:300–301; author reply 300-301, 2004. PubMed/NCBI
33	Xu JW, Gong J, Chang XM, Luo JY, Dong L, Jia A and Xu GP: Effects of estradiol on liver estrogen receptor-alpha and its mRNA expression in hepatic fibrosis in rats. World J Gastroenterol. 10:250–254. 2004. View Article : Google Scholar : PubMed/NCBI
34	Li JF, Chen BC, Lai DD, Jia ZR, Andersson R, Zhang B, Yao JG and Yu Z: Soy isoflavone delays the progression of thioacetamide-induced liver fibrosis in rats. Scand J Gastroenterol. 46:341–349. 2011. View Article : Google Scholar
35	Lee ES, Shin MO, Yoon S and Moon JO: Resveratrol inhibits dimethylnitrosamine-induced hepatic fibrosis in rats. Arch Pharm Res. 33:925–932. 2010. View Article : Google Scholar : PubMed/NCBI
36	Demiroren K, Dogan Y, Kocamaz H, Ozercan IH, Ilhan S, Ustundag B and Bahcecioglu IH: Protective effects of L-carnitine, N-acetylcysteine and genistein in an experimental model of liver fibrosis. Clin Res Hepatol Gastroenterol. 38:63–72. 2014. View Article : Google Scholar
37	Novo E, Busletta C, Bonzo LV, Povero D, Paternostro C, Mareschi K, Ferrero I, David E, Bertolani C, Caligiuri A, et al: Intracellular reactive oxygen species are required for directional migration of resident and bone marrow-derived hepatic pro-fibrogenic cells. J Hepatol. 54:964–974. 2011. View Article : Google Scholar
38	Marra F, Arrighi MC, Fazi M, Caligiuri A, Pinzani M, Romanelli RG, Efsen E, Laffi G and Gentilini P: Extracellular signal-regulated kinase activation differentially regulates platelet-derived growth factor's actions in hepatic stellate cells, and is induced by in vivo liver injury in the rat. Hepatology. 30:951–958. 1999. View Article : Google Scholar : PubMed/NCBI
39	Zhong W, Shen WF, Ning BF, Hu PF, Lin Y, Yue HY, Yin C, Hou JL, Chen YX, Zhang JP, et al: Inhibition of extracellular signal-regulated kinase 1 by adenovirus mediated small interfering RNA attenuates hepatic fibrosis in rats. Hepatology. 50:1524–1536. 2009. View Article : Google Scholar : PubMed/NCBI
40	Frémin C, Bessard A, Ezan F, Gailhouste L, Régeard M, Le Seyec J, Gilot D, Pagès G, Pouysségur J, Langouët S, et al: Multiple division cycles and long-term survival of hepatocytes are distinctly regulated by extracellular signal-regulated kinases ERK1 and ERK2. Hepatology. 49:930–939. 2009. View Article : Google Scholar : PubMed/NCBI
41	Kluwe J, Pradere JP, Gwak GY, Mencin A, De Minicis S, Österreicher CH, Colmenero J, Bataller R and Schwabe RF: Modulation of hepatic fibrosis by c-Jun-N-terminal kinase inhibition. Gastroenterology. 138:347–359. 2010. View Article : Google Scholar :
42	Zhou Y, Shimizu I, Lu G, Itonaga M, Okamura Y, Shono M, Honda H, Inoue S, Muramatsu M and Ito S: Hepatic stellate cells contain the functional estrogen receptor beta but not the estrogen receptor alpha in male and female rats. Biochem Biophys Res Commun. 286:1059–1065. 2001. View Article : Google Scholar : PubMed/NCBI
43	Li Y, Liu J, Lin L, Que R, Shen Y and Tao Z: Regulation of saikosaponin-d on the level of of ERα and ERβ protein in hepatic stellate cells. Trad Chin Drug Res Clin Pharmacol. 27:58–61. 2016.In Chinese.
44	Nadal- Serrano M, Pons DG, Sastre- Serra J, Blanquer-Rosselló MM, Roca P and Oliver J: Genistein modulates oxidative stress in breast cancer cell lines according to ERα/ERβ ratio: Effects on mitochondrial functionality, sirtuins, uncoupling protein 2 and antioxidant enzymes. Int J Biochem Cell Biol. 45:2045–2051. 2013. View Article : Google Scholar
45	Giddabasappa A, Bauler M, Yepuru M, Chaum E, Dalton JT and Eswaraka J: 17-β estradiol protects ARPE-19 cells from oxidative stress through estrogen receptor-β. Invest Ophthalmol Vis Sci. 51:5278–5287. 2010. View Article : Google Scholar : PubMed/NCBI
46	Ortmann J, Veit M, Zingg S, Di Santo S, Traupe T, Yang Z, Völzmann J, Dubey RK, Christen S and Baumgartner I: Estrogen receptor-a but not -β or GPER inhibits high glucose-induced human VSMC proliferation: Potential role of ROS and ERK. J Clin Endocrinol Metab. 96:220–228. 2011. View Article : Google Scholar