Journal of Natural Medicines

, Volume 72, Issue 2, pp 390–398 | Cite as

Berberine protects HK-2 cells from hypoxia/reoxygenation induced apoptosis via inhibiting SPHK1 expression

  • Jianrao LuEmail author
  • Yang Yi
  • Ronghua PanEmail author
  • Chuanfu Zhang
  • Haiyan Han
  • Jie Chen
  • Wenrui Liu
Original Paper


Renal ischemia reperfusion injury (RIRI) refers to the irreversible damage for renal function when blood perfusion is recovered after ischemia for an extended period, which is common in clinical surgeries and has been regarded as a major risk for acute renal failures (ARF) that is accompanied with unimaginably high morbidity and mortality. Hypoxia during ischemia followed by reoxygenation via reperfusion serves as a major event contributing to cell apoptosis, which has been widely accepted as the vital pathogenesis in RIRI. Preventing apoptosis in renal tubular epithelial cell has been considered as effective method for blocking RIRI. In this paper, we established a hypoxia/reoxygenation (H/R) injury model in human proximal tubular epithelial HK-2 cells. Here, we found increased SPHK1 levels in H/R injured HK-2 cells, which could be significantly down regulated after berberine treatment. Berberine has been reported to exert a protective effect on H/R-induced apoptosis of HK-2 cells. So, in our present study, we planned to investigate whether SPHK1 participated in the anti-apoptosis process of berberine in H/R injured HK-2 cells. Our study confirmed the protective effect of berberine against H/R-induced apoptosis in HK-2 cells through promoting cells viability, inhibiting cells apoptosis, and down-regulating p-P38, caspase-3, caspase-9 as well as SPHK1, while up regulating the ratio of Bcl-2/Bax. However, SPHK1 overexpression in HK-2 cells induced severe apoptosis, which can be significantly ameliorated with additional berberine treatment. We concluded that berberine could remarkably prevent H/R-induced apoptosis in HK-2 cells through down-regulating SPHK1 expression levels, and the mechanisms included the suppression of p38 MAPK activation and mitochondrial stress pathways.


H/R-reduced apoptosis HK-2 cells Berberine SPHK1 



The study was supported by the grant 15401930600 from Shanghai Science and Technology Committee.


  1. 1.
    Weight SC, Bell PRF, Nicholson ML (1996) Renal ischemia-reperfusion injury. Br J Surg 83:162–170CrossRefGoogle Scholar
  2. 2.
    Zhou W, Farrar CA, Abe K, Pratt JR, Marsh JE, Wang Y, Stahl GL, Sacks SH (2000) Predominant role for C5b-9 in renal ischemia/reperfusion injury. J Clin Investig 105:1363–1371CrossRefGoogle Scholar
  3. 3.
    Yu Y, Li M, Su N, Zhang Z, Zhao H, Yu H, Xu Y (2016) Honokiol protects against renal ischemia/reperfusion injury via the suppression of oxidative stress, iNOS, inflammation and STAT3 in rats. Mol Med Rep 13:1353–1360CrossRefGoogle Scholar
  4. 4.
    Burne-Taney MJ, Kofler J, Yokota N, Weisfeldt M, Traystman RJ, Rabb H (2003) Acute renal failure after whole body ischemia is characterized by inflammation and T cell-mediated injury. Am J Physiol Renal Physiol 285:F87–F94CrossRefGoogle Scholar
  5. 5.
    Zhang ZX, Wang S, Huang X, Min WP, Sun H, Liu W, Garcia B, Jevnikar AM (2008) NK cells induce apoptosis in tubular epithelial cells and contribute to renal ischemia-reperfusion injury. J Immunol 181:7489–7498CrossRefGoogle Scholar
  6. 6.
    De VB, Matthijsen RA, Wolfs TG, Van Bijnen AA, Heeringa P, Buurman WA (2003) Inhibition of complement factor C5 protects against renal ischemia-reperfusion injury: inhibition of late apoptosis and inflammation. Transplantation 75:375–382CrossRefGoogle Scholar
  7. 7.
    Du C, Wang S, Diao H, Guan Q, Zhong R, Jevnikar AM (2006) Increasing resistance of tubular epithelial cells to apoptosis by shRNA therapy ameliorates renal ischemia-reperfusion injury. Am J Transplant 6:2256–2267CrossRefGoogle Scholar
  8. 8.
    Song H, Han IY, Kim Y, Kim YH, Choi IW, Seo SK, Jung SY, Park S, Kang MS (2015) The NADPH oxidase inhibitor DPI can abolish hypoxia-induced apoptosis of human kidney proximal tubular epithelial cells through Bcl2 up-regulation via ERK activation without ROS reduction. Life Sci 126:69–75CrossRefGoogle Scholar
  9. 9.
    Kim J, Jung KJ, Park KM (2010) Reactive oxygen species differently regulate renal tubular epithelial and interstitial cell proliferation after ischemia and reperfusion injury. Am J Physiol Renal Physiol 298:F1118–F1129CrossRefGoogle Scholar
  10. 10.
    Yu W, Sheng M, Xu R, Yu J, Cui K, Tong J, Shi L, Ren H, Du H (2013) Berberine protects human renal proximal tubular cells from hypoxia/reoxygenation injury via inhibiting endoplasmic reticulum and mitochondrial stress pathways. J Transl Med 11:1–10CrossRefGoogle Scholar
  11. 11.
    Oguz E, Yilmaz Z, Ozbilge H, Baba F, Tabur S, Yerer MB, Hekimoglu A (2015) Effects of melatonin on the serum levels of pro-inflammatory cytokines and tissue injury after renal ischemia reperfusion in rats. Ren Fail 37:1–5CrossRefGoogle Scholar
  12. 12.
    Guan, Qiunong, Nguan, Christopher YC, Du, Caigan (2010) Expression of transforming growth factor-β1 limits renal ischemia-reperfusion injury. Transplantation 89:1320–1327CrossRefGoogle Scholar
  13. 13.
    Jia Z, Qian C, Qin H (2012) Ischemia-induced apoptosis of intestinal epithelial cells correlates with altered integrin distribution and disassembly of F-actin triggered by calcium overload. Biomed Res Int 2012:617539Google Scholar
  14. 14.
    Paugh SW, Paugh BS, Rahmani M, Kapitonov D, Almenara JA, Kordula T, Milstien S, Adams JK, Zipkin RE, Grant S (2008) sphingosine kinase 1, S1P, leukemia. Blood 112:1382–1391CrossRefGoogle Scholar
  15. 15.
    Gude DR, Alvarez SE, Paugh SW, Mitra P, Yu J, Griffiths R, Barbour SE, Milstien S, Spiegel S (2008) Apoptosis induces expression of sphingosine kinase 1 to release sphingosine-1-phosphate as a “come-and-get-me” signal. Faseb J Off Publ Fed Am Soc Exp Biol 22:2629–2638Google Scholar
  16. 16.
    Jin ZQ, Zhang J, Huang Y, Hoover HE, Vessey DA, Karliner JS (2007) A sphingosine kinase 1 mutation sensitizes the myocardium to ischemia/reperfusion injury. Cardiovasc Res 76:41–50CrossRefGoogle Scholar
  17. 17.
    Su D, Cheng Y, Li S, Dai D, Zhang W, Lv M (2017) Sphk1 mediates neuroinflammation and neuronal injury via TRAF2/NF-κB pathways in activated microglia in cerebral ischemia reperfusion. J Neuroimmunol 305:35–41CrossRefGoogle Scholar
  18. 18.
    Yu H, Che X, Xu X, Zheng M, Zhao Y, He W, Yu J, Xiong J, Li W (2013) Insulin protects apoptotic cardiomyocytes from hypoxia/reoxygenation injury through the sphingosine kinase/sphingosine 1-phosphate axis. PLoS One 8:e80644CrossRefGoogle Scholar
  19. 19.
    Iwayama H, Ueda N (2013) Role of mitochondrial Bax, caspases, and MAPKs for ceramide-induced apoptosis in renal proximal tubular cells. Mol Cell Biochem 379:37–42CrossRefGoogle Scholar
  20. 20.
    Lee HT, Otasetlik A, Fu Y, Nasr SH, Emala CW (2004) Differential protective effects of volatile anesthetics against renal ischemia-reperfusion injury in vivo. Anesthesiology 101:1313–1324CrossRefGoogle Scholar
  21. 21.
    Peng J, Ren X, Lan T, Chen Y, Shao Z, Yang C (2016) Renoprotective effects of ursolic acid on ischemia/reperfusion-induced acute kidney injury through oxidative stress, inflammation and the inhibition of STAT3 and NF-κB activities. Mol Med Rep 14:3397–3402CrossRefGoogle Scholar
  22. 22.
    Jang SM, Yee ST, Choi J, Choi MS, Do GM, Jeon SM, Yeo J, Kim MJ, Seo KI, Lee MK (2009) Ursolic acid enhances the cellular immune system and pancreatic beta-cell function in streptozotocin-induced diabetic mice fed a high-fat diet. Int Immunopharmacol 9:113–119CrossRefGoogle Scholar
  23. 23.
    He W, Fang T, Tu P (2009) Research progress on pharmacological activities of echinacoside. China J Chin Materia Med 34:476–479Google Scholar
  24. 24.
    Wang Y, Hu Z, Sun B, Xu J, Jiang J, Luo M (2015) Ginsenoside Rg3 attenuates myocardial ischemia/reperfusion injury via Akt/endothelial nitric oxide synthase signaling and the B-cell lymphoma/B-cell lymphoma-associated X protein pathway. Mol Med Rep 11:4518–4524CrossRefGoogle Scholar
  25. 25.
    Tan S, Wang G, Guo Y, Gui D, Wang N (2013) Preventive effects of a natural anti-inflammatory agent, astragaloside IV, on ischemic acute kidney injury in rats. Evid Based Complement Altern Med 2013:284025Google Scholar
  26. 26.
    Liu CM, Ma JQ, Sun YZ (2012) Puerarin protects rat kidney from lead-induced apoptosis by modulating the PI3K/Akt/eNOS pathway. Toxicol Appl Pharmacol 258:330CrossRefGoogle Scholar
  27. 27.
    Zhu M, Hospital HC (2016) Study on the effects and the mechanism of puerarin on oxidative stress injury induced by renal ischemia-reperfusion in rats. Mod J Integr Tradit Chin West Med 6:585–589Google Scholar
  28. 28.
    Feng L, Ke N, Cheng F, Guo Y, Li S, Li Q, Li Y (2011) The protective mechanism of ligustrazine against renal ischemia/reperfusion injury. J Surg Res 166:298–305CrossRefGoogle Scholar
  29. 29.
    Da X, Yong X, Wang J, Zeng J, Li H, Hu Y, Zheng DH, Lin YT (2017) Berberine nanoparticles protects tubular epithelial cells from renal ischemia-reperfusion injury. Oncotarget 8:24154–24162CrossRefGoogle Scholar

Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan KK, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of NephrologySeventh People’s Hospital Affiliated to Shanghai University of Traditional Chinese MedicineShanghaiChina
  2. 2.Department of Nephrology, Jingan District Central Hospital/Jingan BranchHuashan Hospital affiliated to Fudan UniversityShanghaiChina
  3. 3.Department of NephrologyLiyang Hospital of traditional Chinese medicineJiangsu ProvinceChina

Personalised recommendations