Advertisement

Human Cell

, Volume 31, Issue 3, pp 199–209 | Cite as

Acetylshikonin from Zicao ameliorates renal dysfunction and fibrosis in diabetic mice by inhibiting TGF-β1/Smad pathway

  • Zezhao Li
  • Zhen Hong
  • Zhiqing Peng
  • Yongcai Zhao
  • Rusheng Shao
Research Article
  • 85 Downloads

Abstract

Diabetic nephropathy (DN) is the major cause of end-stage renal disease in diabetic patients. Zicao, a well-known Chinese traditional medicine, has attracted much attention due to its beneficial effects in various medical fields. In this study, we attempted to investigate the effects and mechanisms of action of acetylshikonin, the main ingredient of Zicao, on renal dysfunction in DN. Our results showed that administration with acetylshikonin not only decreased blood urea nitrogen, urine creatinine and the mean kidney-to-body weight ratio in streptozotocin-induced diabetic mice, but also restored the loss of body weight, whereas the blood glucose was not changed. Masson’s trichrome staining showed that acetylshikonin treatment resulted in a marked decrease in kidney fibrosis from diabetic mice. The increased expression of fibrosis proteins, such as plasminogen activator inhibitor type 1 (PAI-1), connective tissue growth factor, and collagen III and IV, were reduced after acetylshikonin administration. In addition, the expressions of interleukin-1β, interleukin-6, monocyte chemoattractant protein-1, intercellular adhesion molecule 1 and infiltration of macrophages in kidney tissues were decreased in acetylshikonin-treated diabetic mice. Acetylshikonin led to a reduction of transforming growth factor-β1 (TGF-β1) expression and Smad-2/3 phosphorylation, as accompanied by increased Smad7 expression. Furthermore, in vitro treatment with acetylshikonin markedly attenuated TGF-β1-induced the PAI-1, collagen III and IV, and Smad-2/3 phosphorylation in HK2 immortalized human proximal tubule epithelial cells. Acetylshikonin also prevented epithelial-to-mesenchymal transition induced by TGF-β1. Collectively, our study provides evidences that acetylshikonin attenuates renal fibrosis though inhibiting TGF-β1/Smad signaling pathway, suggesting that acetylshikonin may be a novel therapeutic agent for the treatment of DN.

Keywords

Diabetic nephropathy Acetylshikonin Inflammation Renal fibrosis TGF-β1/Smad 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

13577_2017_192_MOESM1_ESM.docx (91 kb)
Supplementary material 1 (DOCX 90 kb)

References

  1. 1.
    Mochizuki Y, Tanaka H, Matsumoto K, Sano H, Shimoura H, Ooka J, Sawa T, Ryo-Koriyama K, Hirota Y, Ogawa W, Hirata K. Impaired mechanics of left ventriculo-atrial coupling in patients with diabetic nephropathy. Circ J. 2016;80(9):1957–64.  https://doi.org/10.1253/circj.CJ-16-0488.CrossRefPubMedGoogle Scholar
  2. 2.
    Ritz E, Rychlik I, Locatelli F, Halimi S. End-stage renal failure in type 2 diabetes: a medical catastrophe of worldwide dimensions. Am J Kidney Dis. 1999;34(5):795–808.  https://doi.org/10.1016/S0272-6386(99)70035-1.CrossRefPubMedGoogle Scholar
  3. 3.
    Ni WJ, Tang LQ, Zhou H, Ding HH, Qiu YY. Renoprotective effect of berberine via regulating the PGE2-EP1-Galphaq-Ca(2+) signalling pathway in glomerular mesangial cells of diabetic rats. J Cell Mol Med. 2016;20(8):1491–502.  https://doi.org/10.1111/jcmm.12837.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Deckert T, Poulsen JE, Larsen M. Prognosis of diabetics with diabetes onset before the age of thirty-one. I. Survival, causes of death, and complications. Diabetologia. 1978;14(6):363–70.CrossRefPubMedGoogle Scholar
  5. 5.
    Liu Y. Epithelial to mesenchymal transition in renal fibrogenesis: pathologic significance, molecular mechanism, and therapeutic intervention. J Am Soc Nephrol. 2004;15(1):1–12.CrossRefPubMedGoogle Scholar
  6. 6.
    Wang Z, Han Z, Tao J, Wang J, Liu X, Zhou W, Xu Z, Zhao C, Wang Z, Tan R, Gu M. Role of endothelial-to-mesenchymal transition induced by TGF-beta1 in transplant kidney interstitial fibrosis. J Cell Mol Med. 2017;.  https://doi.org/10.1111/jcmm.13157.Google Scholar
  7. 7.
    Matsushita Y, Ogawa D, Wada J, Yamamoto N, Shikata K, Sato C, Tachibana H, Toyota N, Makino H. Activation of peroxisome proliferator-activated receptor delta inhibits streptozotocin-induced diabetic nephropathy through anti-inflammatory mechanisms in mice. Diabetes. 2011;60(3):960–8.  https://doi.org/10.2337/db10-1361.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Navarro-Gonzalez JF, Mora-Fernandez C. The role of inflammatory cytokines in diabetic nephropathy. J Am Soc Nephrol. 2008;19(3):433–42.  https://doi.org/10.1681/ASN.2007091048.CrossRefPubMedGoogle Scholar
  9. 9.
    Sanz AB, Sanchez-Nino MD, Ramos AM, Moreno JA, Santamaria B, Ruiz-Ortega M, Egido J, Ortiz A. NF-kappaB in renal inflammation. J Am Soc Nephrol. 2010;21(8):1254–62.  https://doi.org/10.1681/ASN.2010020218.CrossRefPubMedGoogle Scholar
  10. 10.
    Kawai T, Rakugi H. Which indexes are the most important risk factor for cardiorenal events in type 2 diabetic patients? Circ J. 2013;77(11):2700–1.CrossRefPubMedGoogle Scholar
  11. 11.
    Rosolowsky ET, Skupien J, Smiles AM, Niewczas M, Roshan B, Stanton R, Eckfeldt JH, Warram JH, Krolewski AS. Risk for ESRD in type 1 diabetes remains high despite renoprotection. J Am Soc Nephrol. 2011;22(3):545–53.  https://doi.org/10.1681/ASN.2010040354.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Guo C, Liu Y, Zhao W, Wei S, Zhang X, Wang W, Zeng X. Apelin promotes diabetic nephropathy by inducing podocyte dysfunction via inhibiting proteasome activities. J Cell Mol Med. 2015;19(9):2273–85.  https://doi.org/10.1111/jcmm.12619.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Chennasamudram SP, Kudugunti S, Boreddy PR, Moridani MY, Vasylyeva TL. Renoprotective effects of (+)-catechin in streptozotocin-induced diabetic rat model. Nutr Res. 2012;32(5):347–56.  https://doi.org/10.1016/j.nutres.2012.03.015.CrossRefPubMedGoogle Scholar
  14. 14.
    Yang QH, Liang Y, Xu Q, Zhang Y, Xiao L, Si LY. Protective effect of tetramethylpyrazine isolated from Ligusticum chuanxiong on nephropathy in rats with streptozotocin-induced diabetes. Phytomedicine. 2011;18(13):1148–52.  https://doi.org/10.1016/j.phymed.2011.05.003.CrossRefPubMedGoogle Scholar
  15. 15.
    Su ML, He Y, Li QS, Zhu BH. Efficacy of Acetylshikonin in preventing obesity and hepatic steatosis in db/db mice. Molecules. 2016;.  https://doi.org/10.3390/molecules21080976.Google Scholar
  16. 16.
    Zorman J, Susjan P, Hafner-Bratkovic I. Shikonin suppresses NLRP3 and AIM2 inflammasomes by direct inhibition of caspase-1. PLoS One. 2016;11(7):e0159826.  https://doi.org/10.1371/journal.pone.0159826.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Andujar I, Rios JL, Giner RM, Recio MC. Pharmacological properties of shikonin—a review of literature since 2002. Planta Med. 2013;79(18):1685–97.  https://doi.org/10.1055/s-0033-1350934.CrossRefPubMedGoogle Scholar
  18. 18.
    Zeng Z, Zhu BH. Arnebin-1 promotes the angiogenesis of human umbilical vein endothelial cells and accelerates the wound healing process in diabetic rats. J Ethnopharmacol. 2014;154(3):653–62.  https://doi.org/10.1016/j.jep.2014.04.038.CrossRefPubMedGoogle Scholar
  19. 19.
    Cheng YW, Chang CY, Lin KL, Hu CM, Lin CH, Kang JJ. Shikonin derivatives inhibited LPS-induced NOS in RAW 264.7 cells via downregulation of MAPK/NF-kappaB signaling. J Ethnopharmacol. 2008;120(2):264–71.  https://doi.org/10.1016/j.jep.2008.09.002.CrossRefPubMedGoogle Scholar
  20. 20.
    Su M, Huang W, Zhu B. Acetylshikonin from zicao prevents obesity in rats on a high-fat diet by inhibiting lipid accumulation and inducing lipolysis. PLoS One. 2016;11(1):e0146884.  https://doi.org/10.1371/journal.pone.0146884.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Pan Y, Huang Y, Wang Z, Fang Q, Sun Y, Tong C, Peng K, Wang Y, Miao L, Cai L, Zhao Y, Liang G. Inhibition of MAPK-mediated ACE expression by compound C66 prevents STZ-induced diabetic nephropathy. J Cell Mol Med. 2014;18(2):231–41.  https://doi.org/10.1111/jcmm.12175.CrossRefPubMedGoogle Scholar
  22. 22.
    Ozcan F, Ozmen A, Akkaya B, Aliciguzel Y, Aslan M. Beneficial effect of myricetin on renal functions in streptozotocin-induced diabetes. Clin Exp Med. 2012;12(4):265–72.  https://doi.org/10.1007/s10238-011-0167-0.CrossRefPubMedGoogle Scholar
  23. 23.
    Pohlers D, Brenmoehl J, Loffler I, Muller CK, Leipner C, Schultze-Mosgau S, Stallmach A, Kinne RW, Wolf G. TGF-beta and fibrosis in different organs—molecular pathway imprints. Biochem Biophys Acta. 2009;1792(8):746–56.  https://doi.org/10.1016/j.bbadis.2009.06.004.PubMedGoogle Scholar
  24. 24.
    Chen HY, Huang XR, Wang W, Li JH, Heuchel RL, Chung AC, Lan HY. The protective role of Smad7 in diabetic kidney disease: mechanism and therapeutic potential. Diabetes. 2011;60(2):590–601.  https://doi.org/10.2337/db10-0403.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Tian W, Lei H, Guan R, Xu Y, Li H, Wang L, Yang B, Gao Z, Xin Z. Icariside II ameliorates diabetic nephropathy in streptozotocin-induced diabetic rats. Drug Des Dev Ther. 2015;9:5147–57.  https://doi.org/10.2147/DDDT.S90060.Google Scholar
  26. 26.
    He Y, Li Q, Su M, Huang W, Zhu B. Acetylshikonin from Zicao exerts antifertility effects at high dose in rats by suppressing the secretion of GTH. Biochem Biophys Res Commun. 2016;476(4):560–5.  https://doi.org/10.1016/j.bbrc.2016.05.162.CrossRefPubMedGoogle Scholar
  27. 27.
    Kim DJ, Lee JH, Park HR, Choi YW. Acetylshikonin inhibits growth of oral squamous cell carcinoma by inducing apoptosis. Arch Oral Biol. 2016;70:149–57.  https://doi.org/10.1016/j.archoralbio.2016.06.020.CrossRefPubMedGoogle Scholar
  28. 28.
    Vaughan DE, Rai R, Khan SS, Eren M, Ghosh AK. Plasminogen activator inhibitor-1 is a marker and a mediator of senescence. Arterioscler Thromb Vasc Biol. 2017;.  https://doi.org/10.1161/ATVBAHA.117.309451.PubMedGoogle Scholar
  29. 29.
    Weston BS, Wahab NA, Mason RM. CTGF mediates TGF-beta-induced fibronectin matrix deposition by upregulating active alpha5beta1 integrin in human mesangial cells. J Am Soc Nephrol. 2003;14(3):601–10.CrossRefPubMedGoogle Scholar
  30. 30.
    Wolf G. Growth factors and the development of diabetic nephropathy. Curr Diab Rep. 2003;3(6):485–90.CrossRefPubMedGoogle Scholar
  31. 31.
    Sonnylal S, Shi-Wen X, Leoni P, Naff K, Van Pelt CS, Nakamura H, Leask A, Abraham D, Bou-Gharios G, de Crombrugghe B. Selective expression of connective tissue growth factor in fibroblasts in vivo promotes systemic tissue fibrosis. Arthritis Rheum. 2010;62(5):1523–32.  https://doi.org/10.1002/art.27382.CrossRefPubMedGoogle Scholar
  32. 32.
    Liu Y. New insights into epithelial-mesenchymal transition in kidney fibrosis. J Am Soc Nephrol. 2010;21(2):212–22.  https://doi.org/10.1681/ASN.2008121226.CrossRefPubMedGoogle Scholar
  33. 33.
    Lan HY. Transforming growth factor-beta/Smad signalling in diabetic nephropathy. Clin Exp Pharmacol Physiol. 2012;39(8):731–8.  https://doi.org/10.1111/j.1440-1681.2011.05663.x.CrossRefPubMedGoogle Scholar
  34. 34.
    Kavsak P, Rasmussen RK, Causing CG, Bonni S, Zhu H, Thomsen GH, Wrana JL. Smad7 binds to Smurf2 to form an E3 ubiquitin ligase that targets the TGF beta receptor for degradation. Mol Cell. 2000;6(6):1365–75.CrossRefPubMedGoogle Scholar
  35. 35.
    Hong SW, Isono M, Chen S, Iglesias-De La Cruz MC, Han DC, Ziyadeh FN. Increased glomerular and tubular expression of transforming growth factor-beta1, its type II receptor, and activation of the Smad signaling pathway in the db/db mouse. Am J Pathol. 2001;158(5):1653–63.CrossRefPubMedGoogle Scholar

Copyright information

© Japan Human Cell Society and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  • Zezhao Li
    • 1
  • Zhen Hong
    • 2
  • Zhiqing Peng
    • 3
  • Yongcai Zhao
    • 4
  • Rusheng Shao
    • 2
  1. 1.Department of Traditional Chinese MedicineCangzhou Central HospitalCangzhouPeople’s Republic of China
  2. 2.Department of NeurologyCangzhou Central HospitalHebeiPeople’s Republic of China
  3. 3.Department of Education AdministrationCangzhou Central HospitalHebeiPeople’s Republic of China
  4. 4.Department of EndocrinologyCangzhou Central HospitalHebeiPeople’s Republic of China

Personalised recommendations