Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 392, Issue 9, pp 1169–1180 | Cite as

Salvianolic acid B protects against ANIT-induced cholestatic liver injury through regulating bile acid transporters and enzymes, and NF-κB/IκB and MAPK pathways

  • Shengnan Li
  • Rong Wang
  • Bin Wu
  • Yuanyuan Wang
  • Fuxing Song
  • Yanqiu Gu
  • Yongfang YuanEmail author
Original Article


The purpose of this study was to investigate the pharmacological effects of salvianolic acid B (SA-B) on α-naphthylisothiocyanate (ANIT)-induced cholestatic liver injury with the focus on bile acid homeostasis and anti-inflammatory pathways. Rats were randomly assigned into four groups. The control group was given normal saline (i.p.) for 7 consecutive days and on the 5th day was given the vehicle (i.g.). Model group was treated with normal saline (i.p.) for 7 days and administrated with ANIT (75 mg/kg, i.g.) on the 5th day. The SA-B groups were treated with SA-B (15 mg/kg and 30 mg/kg, i.p.) for 7 consecutive days as well as ANIT (75 mg/kg, i.g.) on the 5th day. We found that the serum levels of ALT, γ-GT, TBA, and other liver function indexes were found to be lower in the SA-B treatment groups than in the model group. SA-B also upregulated the transporters and enzymes involved in bile acid homeostasis such as Bsep, Oatp2, and Cyp3a2 in rats and BSEP, CYP3A4, and OATP2 in human cell lines. Moreover, SA-B suppressed NF-κB translocation into the nucleus, inhibited phosphorylation of p38 and JNK, and inhibited inflammation markers including IL-1β, IL-6, TGF-β, TNF-α, and COX-2 to extenuate cholestatic liver injury both in vivo and vitro. Taken together, our findings suggest that anti-cholestatic effects of SA-B may be associated with its ability to regulate NF-κB/IκB and MAPK inflammatory signaling pathways to inhibit inflammation and regulate transporters and enzymes to maintain bile acid homeostasis.


Cholestatic liver injury SA-B Inflammation Bile acid homeostasis 



Salvianolic acid B




Bile salt export pump


Multidrug resistance-associate protein 3


Cytochrome P450 3A4


Uridine diphosphate-5′-glucuronosyltransferase 1A1






Tumor necrosis factor-α


Organic anion transporting polypeptide 2


Transforming growth factor-β


Cyclooxygenase 2


Alanine aminotransferase


Aspartate aminotransferase


Cytochrome P450 3a2


Alkaline phosphatase


γ-Glutamyl transferase


Direct bilirubin


Total bilirubin


Total bile acid


Authors’ contributions

S.L., R.W. and F.Y. conceived and designed the research. S.L. and B.W. conducted all experiments. Y.W. and R.W. gave comments in the experimental design and manuscript. F.S. and Y.G. carried out data interpretation and discussion. S.L., R.W., and F.Y. wrote the manuscript. All authors read and approved the manuscript.


This work was financially supported by the National Natural Science Foundation of China (81803815) and the Fund of Shanghai Science and Technology Committee (17401900800).

Compliance with ethical standards

All animal procedures in this work were conducted according to the Animal Ethics Committee, Shanghai 9th People’s Hospital, Shanghai Jiao Tong University School of Medicine, China.

Conflict of interest

The authors declare that they have no conflicts of interest.


  1. Allen K, Jaeschke H, Copple BL (2011) Bile acids induce inflammatory genes in hepatocytes : a novel mechanism of inflammation during Obstructive Cholestasis. Am J Pathol 178:175–186CrossRefGoogle Scholar
  2. Baek HS, Park N, Kwon YJ, Ye DJ, Shin S, Chun YJ (2017) Annexin A5 suppresses cyclooxygenase-2 expression by downregulating the protein kinase C-ζ-nuclear factor-κB signaling pathway in prostatfe cancer cells. Oncotarget 8:74263–74275Google Scholar
  3. Cao W, Guo XW, Zheng HZ, Li DP, Jia GB, Wang J (2012) Current progress of research on pharmacologic actions of salvianolic acid B. Chin J Integr Med 18:316–320CrossRefGoogle Scholar
  4. Chen WY, Lin SY, Pan HC, Liao SL, Chuang YH, Yen YJ, Lin SY, Chen CJ (2012) Beneficial effect of docosahexaenoic acid on cholestatic liver injury in rats. J Nutr Biochem 23:252–264CrossRefGoogle Scholar
  5. Cheng Y, Woolf TF, Gan J, He K (2016) In vitro model systems to investigate bile salt export pump (BSEP) activity and drug interactions: a review. Chem Biol Interact 255:23–30CrossRefGoogle Scholar
  6. Cingolani F, Czaja M (2018) Oxidized albumin - a Trojan horse for p38 MAPK-mediated inflammation in decompensated cirrhosis. Hepatology 68:1678–1680CrossRefGoogle Scholar
  7. Donepudi AC, Aleksunes LM, Driscoll MV, Seeram NP, Slitt AL (2012) The traditional ayurvedic medicine, Eugenia jambolana (Jamun fruit), decreases liver inflammation, injury and fibrosis during cholestasis. Liver Int 32:560–573CrossRefGoogle Scholar
  8. Dong Z, Su L, Esmaili S, Iseli TJ, Ramezani-Moghadam M, Hu L, Xu A, George J, Wang J (2015) Adiponectin attenuates liver fibrosis by inducing nitric oxide production of hepatic stellate cells. J Mol Med 93:1327–1339CrossRefGoogle Scholar
  9. Gäbele E, Froh M, Arteel GE, Uesugi T, Hellerbrand C, Schölmerich J, Brenner DA, Thurman RG, Rippe RA (2009) TNFα is required for cholestasis-induced liver fibrosis in the mouse. Biochem Biophys Res Commun 378:348–353CrossRefGoogle Scholar
  10. Geier A, Wagner M, Dietrich CG, Trauner M (2007) Principles of hepatic organic anion transporter regulation during cholestasis, inflammation and liver regeneration. Biochim Biophys Acta 1773:283–308CrossRefGoogle Scholar
  11. Gowert NS, Klier M, Reich M, Reusswig F, Donner L, Keitel V, Häussinger D, Elvers M (2017) Defective platelet activation and bleeding complications upon cholestasis in mice. Cell Physiol Biochem 41:2133–2149CrossRefGoogle Scholar
  12. Higuchi H, Grambihler A, Canbay A, Bronk SF, Gores GJ (2004) Bile acids up-regulate death receptor 5/TRAIL-receptor 2 expression via a c-Jun N-terminal kinase-dependent pathway involving Sp1. J Biol Chem 279:51–60CrossRefGoogle Scholar
  13. Hirohashi T, Suzuki H, Takikawa H, Sugiyama Y (2000) ATP-dependent transport of bile salts by rat multidrug resistance-associated protein 3 (Mrp3). J Biol Chem 275:2905–2910CrossRefGoogle Scholar
  14. Hirschfield GM, Chapman RW, Karlsen TH, Lammert F, Lazaridis KN, Mason AL (2013) The genetics of complex cholestatic disorders. Gastroenterology 144:1357–1374CrossRefGoogle Scholar
  15. Ho HC, Hong CY (2011) Salvianolic acids: small compounds with multiple mechanisms for cardiovascular protection. J Biomed Sci 18:30–30CrossRefGoogle Scholar
  16. Hong M, Li S, Wang N, Tan HY, Fan C, Feng Y (2017) 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 18:620CrossRefGoogle Scholar
  17. Jin F et al (2013) Anti-inflammatory and anti-oxidative effects of corilagin in a rat model of acute cholestasis. BMC Gastroenterol 13:1–10CrossRefGoogle Scholar
  18. Kerb R, Hoffmeyer S, Brinkmann U (2001) ABC drug transporters: hereditary polymorphisms and pharmacological impact in MDR1, MRP1 and MRP2. Pharmacogenomics 2:51–64CrossRefGoogle Scholar
  19. Kim EK, Choi EJ (2010) Pathological roles of MAPK signaling pathways in human diseases. Biochim Biophys Acta 1802:396–405CrossRefGoogle Scholar
  20. Krajarng A, Imoto M, Tashiro E, Fujimaki T, Shinjo S, Watanapokasin R (2015) Apoptosis induction associated with the ER stress response through up-regulation of JNK in HeLa cells by gambogic acid. BMC Complement Altern Med 15:26CrossRefGoogle Scholar
  21. Kyriakis JM, Avruch J (2001) Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev 81:807–869CrossRefGoogle Scholar
  22. Lam P, Wang R, Ling V (2005) Bile acid transport in sister of P-glycoprotein (ABCB11) knockout mice. Biochemistry 44:12598CrossRefGoogle Scholar
  23. Li D, Zimmerman TL, Thevananther S, Lee HY, Kurie JM, Karpen SJ (2002) Interleukin-1 beta-mediated suppression of RXR:RAR transactivation of the Ntcp promoter is JNK-dependent. J Biol Chem 277:31416–31422CrossRefGoogle Scholar
  24. Li YR, Lin CC, Huang CY, Wong YH, Hsieh CH, Wu HW, Chen JJW, Wu YS (2017) Study of the inhibitory effects on TNF-α-induced NF-κB activation of IMD0354 analogs. Chem Biol Drug Des 90:1307–1311CrossRefGoogle Scholar
  25. Liu YQ, Yuan LM, Gao ZZ, Xiao YS, Sun HY, Yu LS, Zeng S (2016) Dimerization of human uridine diphosphate glucuronosyltransferase allozymes 1A1 and 1A9 alters their quercetin glucuronidation activities. Sci Rep 6:23763CrossRefGoogle Scholar
  26. Liu Q, Shi X, Tang L, Xu W, Jiang S, Ding W, Feng Q, Chu H, Ma Y, Li Y, Lu J, Pu W, Zhou X, Jin L, Wang J, Wu W (2018) Salvianolic acid B attenuates experimental pulmonary inflammation by protecting endothelial cells against oxidative stress injury. Eur J Pharmacol 840:9–19CrossRefGoogle Scholar
  27. Meyer UA (1996) Overview of enzymes of drug metabolism. J Pharmacokinet Biopharm 24:449–459CrossRefGoogle Scholar
  28. Miyake JH, Wang SL, Davis RA (2000) Bile acid induction of cytokine expression by macrophages correlates with repression of hepatic cholesterol 7alpha-hydroxylase. J Biol Chem 275:21805–21808CrossRefGoogle Scholar
  29. Napetschnig J, Wu H (2013) Molecular basis of NF-κB signaling. Annu Rev Biophys 42:443CrossRefGoogle Scholar
  30. Ou QQ, Qian XH, Li DY, Zhang YX, Pei XN, Chen JW, Yu L (2015) Yinzhihuang attenuates ANIT-induced intrahepatic cholestasis in rats through upregulation of Mrp2 and Bsep expressions. Pediatr Res 79:589CrossRefGoogle Scholar
  31. Poupko R, Müller K, Krieger C, Zimmermann H, Luz Z (2004) The two NF-kappa B activation pathways and their role in innate and adaptive immunity. Trends Immunol 25:280–288CrossRefGoogle Scholar
  32. Qi Z et al (2015) Sodium houttuyfonate inhibits inflammation by blocking the MAPKs/NF-κB signaling pathways in bovine endometrial epithelial cells. Res Vet Sci 100:245–251CrossRefGoogle Scholar
  33. Rong W, Hai Z, Wang Y, Song F, Yuan Y (2017) Inhibitory effects of quercetin on the progression of liver fibrosis through the regulation of NF-кB/IкBα, p38 MAPK, and Bcl-2/Bax signaling. Int Immunopharmacol 47:126–133CrossRefGoogle Scholar
  34. Sakurai M, Saito F, Ohata Y, Yabe Y, Nishi T (2011) Organic anion transporting polypeptides (OATPs): regulation of expression and function. Curr Drug Metab 12:139CrossRefGoogle Scholar
  35. Shelby MK, Cherrington NJ, Vansell NR, Klaassen CD (2003) Tissue mRNA expression of the rat UDP-glucuronosyltransferase gene family. Drug Metab Dispos 31:326–333CrossRefGoogle Scholar
  36. Slopianka M, Herrmann A, Pavkovic M, Ellinger-Ziegelbauer H, Ernst R, Mally A, Keck M, Riefke B (2017) Quantitative targeted bile acid profiling as new markers for DILI in a model of methapyrilene-induced liver injury in rats. Toxicology 386:1–10CrossRefGoogle Scholar
  37. Stieger B (2011) The role of the sodium-taurocholate cotransporting polypeptide (NTCP) and of the bile salt export pump (BSEP) in physiology and pathophysiology of bile formation. Handb Exp Pharmacol 201:205–259CrossRefGoogle Scholar
  38. Sun J, Nan G (2016) The mitogen-activated protein kinase (MAPK) signaling pathway as a discovery target in stroke. J Mol Neurosci 59:1–1CrossRefGoogle Scholar
  39. Trauner M, Meier PJ, Boyer JL (1998) Molecular pathogenesis of cholestasis. N Engl J Med 339:1217–1227CrossRefGoogle Scholar
  40. Wang R, Yu XY, Guo ZY, Wang YJ, Wu Y, Yuan YF (2012) Inhibitory effects of salvianolic acid B on CCl(4)-induced hepatic fibrosis through regulating NF-κB/IκBα signaling. J Ethnopharmacol 144:592–598CrossRefGoogle Scholar
  41. Wang D, Qiao J, Zhao X, Chen T, Guan D (2015a) Thymoquinone inhibits IL-1β-induced inflammation in human osteoarthritis chondrocytes by suppressing NF-κB and MAPKs signaling pathway. Inflammation 38:2235–2241CrossRefGoogle Scholar
  42. Wang Y, Wang R, Wang Y, Peng R, Wu Y, Yuan Y (2015b) Ginkgo biloba extract mitigates liver fibrosis and apoptosis by regulating p38 MAPK, NF-κB/IκBα, and Bcl-2/Bax signaling. Drug Des Dev Ther 9:6303–6317Google Scholar
  43. Wang LL, Wu GX, Wu FH, Jiang N, Lin Y (2017) Geniposide attenuates ANIT-induced cholestasis through regulation oftransporters and enzymes involved in bile acids homeostasis in rat. J Ethnopharmacol 196:178–185CrossRefGoogle Scholar
  44. Wang R, Wang J, Song F, Li S, Yuan Y (2018) Tanshinol ameliorates CCl4-induced liver fibrosis in rats through the regulation of Nrf2/HO-1 and NF-κB/IκBα signaling pathway. Drug Des Dev Ther 12:1281–1292CrossRefGoogle Scholar
  45. Weerachayaphorn J, Luo Y, Mennone A, Soroka CJ, Harry K, Boyer JL (2014) Deleterious effect of oltipraz on extrahepatic cholestasis in bile duct-ligated mice. J Hepatol 60:160–166CrossRefGoogle Scholar
  46. Weston CR, Davis RJ (2002) The JNK signal transduction pathway. Curr Opin Genet Dev 12:14–21CrossRefGoogle Scholar
  47. Woolbright BL, Antoine DJ, Jenkins RE, Bajt ML, Park BK, Jaeschke H (2013) Plasma biomarkers of liver injury and inflammation demonstrate a lack of apoptosis during obstructive cholestasis in mice. Toxicol Appl Pharmacol 273:524–531CrossRefGoogle Scholar
  48. Xia ZB, Yuan YJ, Zhang QH, Li H, Dai JL, Min JK (2018) Salvianolic acid B suppresses inflammatory mediator levels by downregulating NF-κB in a rat model of rheumatoid arthritis. Med Sci Monit 24:2524–2532CrossRefGoogle Scholar
  49. Xie W, Radominska-Pandya A, Shi Y, Simon CM, Nelson MC, Ong ES, Waxman DJ, Evans RM (2001) An essential role for nuclear receptors SXR/PXR in detoxification of cholestatic bile acids. Proc Natl Acad Sci U S A 98:3375–3380CrossRefGoogle Scholar
  50. Xu L, Sheng T, Liu X, Zhang T, Wang Z, Han H (2017) Analyzing the hepatoprotective effect of the Swertia cincta Burkillextract against ANIT-induced cholestasis in rats by modulating the expression of transporters and metabolic enzymes. J Ethnopharmacol 209:91–99CrossRefGoogle Scholar
  51. Yan JY, Ai G, Zhang XJ, Xu HJ, Huang ZM (2015) Investigations of the total flavonoids extracted from flowers of Abelmoschus manihot (L.) medic against α-naphthylisothiocyanate-induced cholestatic liver injury in rats. J Ethnopharmacol 172:202–213CrossRefGoogle Scholar
  52. Yang T, Mei H, Xu D, Zhou W, Zhu X, Sun L, Huang X, Wang X, Shu T, Liu J, Ding J, Hassan HM, Zhang L, Jiang Z (2017) Early indications of ANIT-induced cholestatic liver injury: alteration of hepatocyte polarization and bile acid homeostasis. Food and chemical toxicology: an international journal published for the. Br Ind Biol Res Assoc 110:1–12. Google Scholar
  53. Yu F, Lu Z, Chen B, Wu X, Dong P, Zheng J (2015) Salvianolic acid B-induced microRNA-152 inhibits liver fibrosis by attenuating DNMT1-mediated Patched1 methylation. J Cell Mol Med 19:2617–2632CrossRefGoogle Scholar
  54. Yue S, Hu B, Wang Z, Yue Z, Wang F, Zhao Y, Yang Z, Shen M (2014) Salvia miltiorrhiza compounds protect the liver from acute injury by regulation of p38 and NFκB signaling in Kupffer cells. Pharm Biol 52:1278–1285CrossRefGoogle Scholar
  55. Zeng H, Jiang Y, Chen P, Fan X, Li D, Liu A, Ma X, Xie W, Liu P, Gonzalez FJ, Huang M, Bi H (2017) Schisandrol B protects against cholestatic liver injury through pregnane X receptors. Br J Pharmacol 174:672–688CrossRefGoogle Scholar
  56. Zhao Y, He X, Ma X, Wen J, Li P, Wang J, Li R, Zhu Y, Wei S, Li H, Zhou X, Li K, Liu H, Xiao X (2017) Paeoniflorin ameliorates cholestasis via regulating hepatic transporters and suppressing inflammation in ANIT-fed rats. Biomed Pharmacother 89:61–68CrossRefGoogle Scholar
  57. Zhou F, Xu X, Wang D, Wu J, Wang J (2017) Identification of novel NF-ĸB transcriptional targets in TNFα-treated HeLa and HepG2 cells. Cell Biol Int 41:555–569CrossRefGoogle Scholar
  58. Zhou Y et al (2018) SB203580 attenuates acute lung injury and inflammation in rats with acute pancreatitis in pregnancy. InflammopharmacologyGoogle Scholar
  59. Zollner G, Marschall HU, Wagner M, Trauner M (2006) Role of nuclear receptors in the adaptive response to bile acids and cholestasis: pathogenetic and therapeutic considerations. Mol Pharm 3:231–251CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Shengnan Li
    • 1
  • Rong Wang
    • 1
  • Bin Wu
    • 1
  • Yuanyuan Wang
    • 1
  • Fuxing Song
    • 1
  • Yanqiu Gu
    • 1
  • Yongfang Yuan
    • 1
    Email author
  1. 1.Department of Pharmacy, Shanghai 9th People’s HospitalShanghai Jiao Tong University School of MedicineShanghaiChina

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