Differential Inhibitory Effects of Salvianolic Acids on Activation of Rat Hepatic Stellate Cells by Platelet-Derived Growth Factor

Abstract

Platelet-derived growth factor (PDGF) induces cell proliferation together with oxidative stress. The present study investigated the effects of salvianolic acid A (Sal A) and B (Sal B) on the PDGF-induced signaling cascades in hepatic stellate cells (HSCs). HSC-T6, a rat hepatic stellate cell line, was stimulated with PDGF (10 ng/mL). The inhibitory effects of Sal A and B on oxidative stress-related signaling pathways were assessed in vitro. The protein levels were measured by Western blotting. FACS analysis was applied to detect the thioredoxin (Trx) level. Sal A and B showed different inhibitory abilities on the PDGF-related pathway. Sal A inhibited 70-kDa ribosomal S6 kinase (p70^s6k) and associated proteins. Sal B attenuated PDGF-induced c-jun-N-terminal kinase (JNK), p38, and PKC-δ phosphorylations. Both Sal A and B diminished the activation of PKD, Trx, heme-oxygenase (HO)-1, and Nrf2. Taken together, our results showed that Sal A and B attenuated PDGF-induced ROS formation in HSCs, possibly through different signaling pathways.

Key words

hepatic stellate cells - salvianolic acid - platelet-derived growth factor - oxidative stress - hepatic fibrosis - Salvia miltiorrhiza - Lamiaceae

References

• 1 Friedman S L. Hepatic stellate cells: protean, multifunctional, and enigmatic cells of the liver.  Physiol Rev. 2008;  88 125-172
• 2 Friedman S L. Mechanisms of hepatic fibrogenesis.  Gastroenterology. 2008;  134 1655-1669
• 3 Adachi T, Togashi H, Suzuki A, Kasai S, Ito J, Sugahara K, Kawata S. NAD(P)H oxidase plays a crucial role in PDGF-induced proliferation of hepatic stellate cells.  Hepatology. 2005;  41 1272-1281
• 4 Griner E M, Kazanietz M G. Protein kinase C and other diacylglycerol effectors in cancer.  Nat Rev Cancer. 2007;  7 281-294
• 5 Fan C Y, Katsuyama M, Yabe-Nishimura C. PKCdelta mediates up-regulation of NOX1, a catalytic subunit of NADPH oxidase, via transactivation of the EGF receptor: possible involvement of PKCdelta in vascular hypertrophy.  Biochem J. 2005;  390 761-767
• 6 Ha K S, Exton J H. Differential translocation of protein kinase C isozymes by thrombin and platelet-derived growth factor. A possible function for phosphatidylcholine-derived diacylglycerol.  J Biol Chem. 1993;  268 10534-10539
• 7 Mitchell F E, Marais R M, Parker P J. The phosphorylation of protein kinase C as a potential measure of activation.  Biochem J. 1989;  261 131-136
• 8 Johannes F J, Prestle J, Eis S, Oberhagemann P, Pfizenmaier K. PKCu is a novel, atypical member of the protein kinase C family.  J Biol Chem. 1994;  269 6140-6148
• 9 Rozengurt E, Rey O, Waldron R T. Protein kinase D signaling.  J Biol Chem. 2005;  280 13205-13208
• 10 Zugaza J L, Waldron R T, Sinnett-Smith J, Rozengurt E. Bombesin, vasopressin, endothelin, bradykinin, and platelet-derived growth factor rapidly activate protein kinase D through a protein kinase C-dependent signal transduction pathway.  J Biol Chem. 1997;  272 23952-23960
• 11 Gebhardt R. Oxidative stress, plant-derived antioxidants and liver fibrosis.  Planta Med. 2002;  68 289-296
• 12 Lee T F, Lin Y L, Huang Y T. Studies on antiproliferative effects of phthalides from Ligusticum chuanxiong in hepatic stellate cells.  Planta Med. 2007;  73 527-534
• 13 Lin Y L, Wu C F, Huang Y T. Phenols from the roots of Rheum palmatum attenuate chemotaxis in rat hepatic stellate cells.  Planta Med. 2008;  74 1246-1252
• 14 Tsai M K, Lin Y L, Huang Y T. Effects of salvianolic acids on oxidative stress and hepatic fibrosis in rats.  Toxicol Appl Pharmacol. 2010;  242 155-164
• 15 Lin Y L, Lee T F, Huang Y J, Huang Y T. Antiproliferative effect of salvianolic acid A on rat hepatic stellate cells.  J Pharm Pharmacol. 2006;  58 933-939
• 16 Lin Y L, Wu C H, Luo M H, Huang Y J, Wang C N, Shiao M S, Huang Y T. In vitro protective effects of salvianolic acid B on primary hepatocytes and hepatic stellate cells.  J Ethnopharmacol. 2006;  105 215-222
• 17 Nilsson J, Soderberg O, Nilsson K, Rosen A. Thioredoxin prolongs survival of B-type chronic lymphocytic leukemia cells.  Blood. 2000;  95 1420-1426
• 18 Kobayashi Y, Bridle K R, Ramm G A, O'Neill R, Britton R S, Bacon B R. Effect of phorbol ester and platelet-derived growth factor on protein kinase C in rat hepatic stellate cells.  Liver Int. 2007;  27 1066-1075
• 19 Liu Y, Wen X M, Lui E L, Friedman S L, Cui W, Ho N P, Li L, Ye T, Fan S T, Zhang H. Therapeutic targeting of the PDGF and TGF-beta-signaling pathways in hepatic stellate cells by PTK787/ZK22258.  Lab Invest. 2009;  89 1152-1160
• 20 Clempus R E, Griendling K K. Reactive oxygen species signaling in vascular smooth muscle cells.  Cardiovasc Res. 2006;  71 216-225
• 21 Bataller R, Schwabe R F, Choi Y H, Yang L, Paik Y H, Lindquist J, Qian T, Schoonhoven R, Hagedorn C H, Lemasters J J, Brenner D A. NADPH oxidase signal transduces angiotensin II in hepatic stellate cells and is critical in hepatic fibrosis.  J Clin Invest. 2003;  112 1383-1394
• 22 Schulze P C, De Keulenaer G W, Yoshioka J, Kassik K A, Lee R T. Vitamin D3-upregulated protein-1 (VDUP-1) regulates redox-dependent vascular smooth muscle cell proliferation through interaction with thioredoxin.  Circ Res. 2002;  91 689-695
• 23 Grogan T M, Fenoglio-Prieser C, Zeheb R, Bellamy W, Frutiger Y, Vela E, Stemmerman G, Macdonald J, Richter L, Gallegos A, Powis G. Thioredoxin, a putative oncogene product, is overexpressed in gastric carcinoma and associated with increased proliferation and increased cell survival.  Hum Pathol. 2000;  31 475-481
• 24 Gabele E, Reif S, Tsukada S, Bataller R, Yata Y, Morris T, Schrum L W, Brenner D A, Rippe R A. The role of p 70S6K in hepatic stellate cell collagen gene expression and cell proliferation.  J Biol Chem. 2005;  280 13374-13382
• 25 Storz P, Toker A. Protein kinase D mediates a stress-induced NF-kappaB activation and survival pathway.  EMBO J. 2003;  22 109-120
• 26 Storz P, Doppler H, Toker A. Protein kinase Cdelta selectively regulates protein kinase D-dependent activation of NF-kappaB in oxidative stress signaling.  Mol Cell Biol. 2004;  24 2614-2626
• 27 Niture S K, Jain A K, Jaiswal A K. Antioxidant-induced modification of INrf2 cysteine 151 and PKC-delta-mediated phosphorylation of Nrf2 serine 40 are both required for stabilization and nuclear translocation of Nrf2 and increased drug resistance.  J Cell Sci. 2009;  122 4452-4464
• 28 Surh Y J. Cancer chemoprevention with dietary phytochemicals.  Nat Rev Cancer. 2003;  3 768-780
• 29 Was H, Cichon T, Smolarczyk R, Rudnicka D, Stopa M, Chevalier C, Leger J J, Lackowska B, Grochot A, Bojkowska K, Ratajska A, Kieda C, Szala S, Dulak J, Jozkowicz A. Overexpression of heme oxygenase-1 in murine melanoma: increased proliferation and viability of tumor cells, decreased survival of mice.  Am J Pathol. 2006;  169 2181-2198
• 30 Huang H C, Nguyen T, Pickett C B. Regulation of the antioxidant response element by protein kinase C-mediated phosphorylation of NF-E2-related factor 2.  Proc Natl Acad Sci USA. 2000;  97 12475-12480
• 31 Churchman A T, Anwar A A, Li F Y, Sato H, Ishii T, Mann G E, Siow R C. Transforming growth factor-beta elicits Nrf2-mediated antioxidant responses in aortic smooth muscle cells.  J Cell Mol Med. 2009;  13 2282-2292
• 32 Malec V, Gottschald O R, Li S, Rose F, Seeger W, Hanze J. HIF-1 alpha signaling is augmented during intermittent hypoxia by induction of the Nrf2 pathway in NOX1-expressing adenocarcinoma A549 cells.  Free Radic Biol Med. 2010;  48 1626-1635
• 33 Catarzi S, Biagioni C, Giannoni E, Favilli F, Marcucci T, Iantomasi T, Vincenzini M T. Redox regulation of platelet-derived-growth-factor-receptor: role of NADPH-oxidase and c-Src tyrosine kinase.  Biochim Biophys Acta. 2005;  1745 166-175