Abstract
Our recent study demonstrated that sodium arsenite at a clinically relevant dose induced nephrotoxicity in human renal proximal tubular epithelial cell line HK-2, which could be inhibited by natural product 2,3,5,6-tetramethylpyrazine (TMP) with antioxidant activity. The present study demonstrated that arsenic exposure resulted in protein and enzymatic induction of heme oxygenase-1 (HO-1) in dose- and time-dependent manners in HK-2 cells. Blocking HO-1 enzymatic activity by zinc protoporphyrin (ZnPP) augmented arsenic-induced apoptosis, ROS production and mitochondrial dysfunction, suggesting a critical role for HO-1 as a renal protectant in this procession. On the other hand, TMP, upstream of HO-1, inhibited arsenic-induced ROS production and ROS-dependent HO-1 expression. TMP also prevented mitochondria dysfunction and suppressed activation of the intrinsic apoptotic pathway in HK-2 cells. Our results revealed that the regulation of arsenic-induced HO-1 expression was performed through multiple ROS-dependent signal pathways and the corresponding transcription factors, including p38 MAPK and JNK (but not ERK), AP-1, Nrf2 and NF-κB. TMP inhibited arsenic-induced activations of JNK, p38 MAPK, ERK, AP-1 and Nrf2 and block HO-1 protein expression. The present study, furthermore, demonstrated arsenic-induced expression of arsenic response protein 2 (ARS2) that was regulated by p38 MAPK, ERK and NF-κB. To our knowledge, this is the first report showing that ARS2 involved in arsenic-induced nephrotoxicity, while TMP pretreatment prevented such an up-regulation of ARS2 in HK-2 cells. Given ARS2 and HO-1 sharing the similar regulation mechanism, we speculated that ARS2 might also mediate cell survival in this procession. In summary, our study highlighted a role of HO-1 in the protection against arsenic-induced cytotoxicity downstream from the primary targets of TMP and further indicated that TMP may be used as a potential therapeutic agent in the treatment of arsenic-induced nephrotoxicity.
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Abbreviations
- AG:
-
Aminoguanidine
- AKI:
-
Acute kidney injury
- ARS2:
-
Arsenic response protein 2
- As:
-
Arsenic
- Bay:
-
Bay 11-7082
- CKD:
-
Chronic kidney disease
- DHE:
-
Dihydroethidium
- FACS:
-
Fluorescence-activated cell sorter
- HO-1:
-
Heme oxygenase-1
- MAPK:
-
Mitogen-activated protein kinase
- Nrf2:
-
Nuclear factor erythroid derived-2
- NAC:
-
N-acetylcysteine
- NF-κB:
-
Nuclear factor-κB
- PARP:
-
Poly ADP-ribose polymerase
- PI:
-
Propidium iodide
- ROS:
-
Reactive oxygen species
- SB:
-
SB203580
- SDH:
-
Sucinate dehydrogenase
- SP:
-
SP600125
- TMP:
-
Tetramethylpyrazine
- TUNEL:
-
Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling
- U0:
-
U0126
- ZnPP:
-
Zinc protoporphyrin
References
Andreu-Agullo C, Maurin T, Thompson CB, Lai EC (2012) Ars2 maintains neural stem-cell identity through direct transcriptional activation of Sox2. Nature 481(7380):195–198. doi:10.1038/nature10712
Billings IF, Yu C, Byrne JG, Petracek MR, Pretorius M (2014) Heme oxygenase-1 and acute kidney injury following cardiac surgery. Cardiorenal Med 4(1):12–21. doi:10.1159/000357871
Chang CF, Liu XM, Peyton KJ, Durante W (2014) Heme oxygenase-1 counteracts contrast media-induced endothelial cell dysfunction. Biochem Pharmacol 87(2):303–311. doi:10.1016/j.bcp.2013.11.002
Chen JW, Chen HY, Li WF, Liou SH, Chen CJ, Wu JH, Wang SL (2011) The association between total urinary arsenic concentration and renal dysfunction in a community-based population from central Taiwan. Chemosphere 84(1):17–24. doi:10.1016/j.chemosphere.2011.02.091
Chen WJ, Huang YL, Shiue HS, Chen TW, Lin YF, Huang CY, Lin YC, Han BC, Hsueh YM (2014) Renin-angiotensin-aldosterone system related gene polymorphisms and urinary total arsenic is related to chronic kidney disease. Toxicol Appl Pharmacol 279(2):95–102. doi:10.1016/j.taap.2014.05.011
Deshane J, Kim J, Bolisetty S, Hock TD, Hill-Kapturczak N, Agarwal A (2010) Sp1 regulates chromatin looping between an intronic enhancer and distal promoter of the human heme oxygenase-1 gene in renal cells. J Biol Chem 285(22):16476–16486. doi:10.1074/jbc.M109.058586
Fan SF, Chao PL, Lin AM (2010) Arsenite induces oxidative injury in rat brain: synergistic effect of iron. Ann N Y Acad Sci 1199:27–35. doi:10.1111/j.1749-6632.2009.05170.x
Garnier N, Petruccelli LA, Molina MF, Kourelis M, Kwan S, Diaz Z, Schipper HM, Gupta A, del Rincon SV, Mann KK, Miller WH Jr (2013) The novel arsenical Darinaparsin circumvents BRG1-dependent, HO-1-mediated cytoprotection in leukemic cells. Leukemia 27(11):2220–2228. doi:10.1038/leu.2013.54
Gong X, Celsi G, Carlsson K, Norgren S, Chen M (2010) N-acetylcysteine amide protects renal proximal tubular epithelial cells against iohexol-induced apoptosis by blocking p38 MAPK and iNOS signaling. Am J Nephrol 31(2):178–188. doi:10.1159/000268161
Gong X, Ivanov VN, Davidson MM, Hei TK (2014) Tetramethylpyrazine (TMP) protects against sodium arsenite-induced nephrotoxicity by suppressing ROS production, mitochondrial dysfunction, pro-inflammatory signaling pathways and programed cell death. Arch Toxicol. doi:10.1007/s00204-014-1302-y
Goodman AI, Olszanecki R, Yang LM, Quan S, Li M, Omura S, Stec DE, Abraham NG (2007) Heme oxygenase-1 protects against radiocontrast-induced acute kidney injury by regulating anti-apoptotic proteins. Kidney Int 72(8):945–953. doi:10.1038/sj.ki.5002447
Gruber JJ, Zatechka DS, Sabin LR, Yong J, Lum JJ, Kong M, Zong WX, Zhang Z, Lau CK, Rawlings J, Cherry S, Ihle JN, Dreyfuss G, Thompson CB (2009) Ars2 links the nuclear cap-binding complex to RNA interference and cell proliferation. Cell 138(2):328–339. doi:10.1016/j.cell.2009.04.046
Hsieh HL, Lin CC, Chan HJ, Yang CM (2014) c-Src-dependent EGF receptor transactivation contributes to ET-1-induced COX-2 expression in brain microvascular endothelial cells. J Neuroinflamm 9:152. doi:10.1186/1742-2094-9-152
Huang JX, Kaeslin G, Ranall MV, Blaskovich MA, Becker B, Butler MS, Little MH, Lash LH, Cooper MA (2015) Evaluation of biomarkers for in vitro prediction of drug-induced nephrotoxicity: comparison of HK-2, immortalized human proximal tubule epithelial, and primary cultures of human proximal tubular cells. Pharmacol Res Perspect 3(3):e00148. doi:10.1002/prp2.148
Ivanov VN, Hei TK (2004) Arsenite sensitizes human melanomas to apoptosis via tumor necrosis factor alpha-mediated pathway. J Biol Chem 279(21):22747–22758. doi:10.1074/jbc.M314131200
Ivanov VN, Hei TK (2005) Combined treatment with EGFR inhibitors and arsenite upregulated apoptosis in human EGFR-positive melanomas: a role of suppression of the PI3K-AKT pathway. Oncogene 24(4):616–626. doi:10.1038/sj.onc.1208125
Ivanov VN, Hei TK (2013) Induction of apoptotic death and retardation of neuronal differentiation of human neural stem cells by sodium arsenite treatment. Exp Cell Res 319(6):875–887. doi:10.1016/j.yexcr.2012.11.019
Kang SH, Song JH, Kang HK, Kang JH, Kim SJ, Kang HW, Lee YK, Park DB (2003) Arsenic trioxide-induced apoptosis is independent of stress-responsive signaling pathways but sensitive to inhibition of inducible nitric oxide synthase in HepG2 cells. Exp Mol Med 35(2):83–90. doi:10.1038/emm.2003.12
Kilic U, Kilic E, Tuzcu Z, Tuzcu M, Ozercan IH, Yilmaz O, Sahin F, Sahin K (2013) Melatonin suppresses cisplatin-induced nephrotoxicity via activation of Nrf-2/HO-1 pathway. Nutr Metab (Lond) 10(1):7. doi:10.1186/1743-7075-10-7
Kiriyama M, Kobayashi Y, Saito M, Ishikawa F, Yonehara S (2009) Interaction of FLASH with arsenite resistance protein 2 is involved in cell cycle progression at S phase. Mol Cell Biol 29(17):4729–4741. doi:10.1128/MCB.00289-09
Lee JE, Park JH, Shin IC, Koh HC (2012) Reactive oxygen species regulated mitochondria-mediated apoptosis in PC12 cells exposed to chlorpyrifos. Toxicol Appl Pharmacol 263(2):148–162. doi:10.1016/j.taap.2012.06.005
Li H, Zhang L, Wang F, Shi Y, Ren Y, Liu Q, Cao Y, Duan H (2011) Attenuation of glomerular injury in diabetic mice with tert-butylhydroquinone through nuclear factor erythroid 2-related factor 2-dependent antioxidant gene activation. Am J Nephrol 33(4):289–297. doi:10.1159/000324694
Li B, Li X, Zhu B, Zhang X, Wang Y, Xu Y, Wang H, Hou Y, Zheng Q, Sun G (2013) Sodium arsenite induced reactive oxygen species generation, nuclear factor (erythroid-2 related) factor 2 activation, heme oxygenase-1 expression, and glutathione elevation in Chang human hepatocytes. Environ Toxicol 28(7):401–410. doi:10.1002/tox.20731
Lim JL, Wilhelmus MM, de Vries HE, Drukarch B, Hoozemans JJ, van Horssen J (2014) Antioxidative defense mechanisms controlled by Nrf2: state-of-the-art and clinical perspectives in neurodegenerative diseases. Arch Toxicol 88(10):1773–1786. doi:10.1007/s00204-014-1338-z
Michael HA (2013) Geochemistry. An arsenic forecast for China. Science 341(6148):852–853. doi:10.1126/science.1242212
Miyagi MY, Seelaender M, Castoldi A, de Almeida DC, Bacurau AV, Andrade-Oliveira V, Enjiu LM, Pisciottano M, Hayashida CY, Hiyane MI, Brum PC, Camara NO, Amano MT (2014) Long-term aerobic exercise protects against cisplatin-induced nephrotoxicity by modulating the expression of IL-6 and HO-1. PLoS ONE 9(10):e108543. doi:10.1371/journal.pone.0108543
Mossman BT, Lounsbury KM, Reddy SP (2006) Oxidants and signaling by mitogen-activated protein kinases in lung epithelium. Am J Respir Cell Mol Biol 34(6):666–669. doi:10.1165/rcmb.2006-0047SF
Peraza MA, Carter DE, Gandolfi AJ (2003) Toxicity and metabolism of subcytotoxic inorganic arsenic in human renal proximal tubule epithelial cells (HK-2). Cell Biol Toxicol 19(4):253–264
Peraza MA, Cromey DW, Carolus B, Carter DE, Gandolfi AJ (2006) Morphological and functional alterations in human proximal tubular cell line induced by low level inorganic arsenic: evidence for targeting of mitochondria and initiated apoptosis. J Appl Toxicol 26(4):356–367. doi:10.1002/jat.1149
Pi J, Diwan BA, Sun Y, Liu J, Qu W, He Y, Styblo M, Waalkes MP (2008) Arsenic-induced malignant transformation of human keratinocytes: involvement of Nrf2. Free Radic Biol Med 45(5):651–658. doi:10.1016/j.freeradbiomed.2008.05.020
Rodriguez-Lado L, Sun G, Berg M, Zhang Q, Xue H, Zheng Q, Johnson CA (2013) Groundwater arsenic contamination throughout China. Science 341(6148):866–868. doi:10.1126/science.1237484
Rossman TG, Wang Z (1999) Expression cloning for arsenite-resistance resulted in isolation of tumor-suppressor fau cDNA: possible involvement of the ubiquitin system in arsenic carcinogenesis. Carcinogenesis 20(2):311–316
Ruiz-Hernandez A, Kuo CC, Rentero-Garrido P, Tang WY, Redon J, Ordovas JM, Navas-Acien A, Tellez-Plaza M (2015) Environmental chemicals and DNA methylation in adults: a systematic review of the epidemiologic evidence. Clin Epigenetics 7(1):55. doi:10.1186/s13148-015-0055-7
Schreiber E, Matthias P, Muller MM, Schaffner W (1989) Rapid detection of octamer binding proteins with ‘mini-extracts’, prepared from a small number of cells. Nucleic Acids Res 17(15):6419
Shen ZX, Chen GQ, Ni JH, Li XS, Xiong SM, Qiu QY, Zhu J, Tang W, Sun GL, Yang KQ, Chen Y, Zhou L, Fang ZW, Wang YT, Ma J, Zhang P, Zhang TD, Chen SJ, Chen Z, Wang ZY (1997) Use of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients. Blood 89(9):3354–3360
Teng YC, Tai YI, Lee YH, Lin AM (2013) Role of HO-1 in the arsenite-induced neurotoxicity in primary cultured cortical neurons. Mol Neurobiol 48(2):281–287. doi:10.1007/s12035-013-8492-9
Wang J, Dore S (2007) Heme oxygenase-1 exacerbates early brain injury after intracerebral haemorrhage. Brain 130(Pt 6):1643–1652
Wang XJ, Sun Z, Chen W, Eblin KE, Gandolfi JA, Zhang DD (2007) Nrf2 protects human bladder urothelial cells from arsenite and monomethylarsonous acid toxicity. Toxicol Appl Pharmacol 225(2):206–213. doi:10.1016/j.taap.2007.07.016
Wang L, Kou MC, Weng CY, Hu LW, Wang YJ, Wu MJ (2012) Arsenic modulates heme oxygenase-1, interleukin-6, and vascular endothelial growth factor expression in endothelial cells: roles of ROS, NF-kappaB, and MAPK pathways. Arch Toxicol 86(6):879–896. doi:10.1007/s00204-012-0845-z
Wang Q, Wu L, Wang J (2013a) Reciprocal regulation of cyclooxygenase 2 and heme oxygenase 1 upon arsenic trioxide exposure in normal human lung fibroblast. J Biochem Mol Toxicol 27(6):323–329. doi:10.1002/jbt.21491
Wang S, Wei Q, Dong G, Dong Z (2013b) ERK-mediated suppression of cilia in cisplatin-induced tubular cell apoptosis and acute kidney injury. Biochim Biophys Acta 1832(10):1582–1590. doi:10.1016/j.bbadis.2013.05.023
Wilson MD, Wang D, Wagner R, Breyssens H, Gertsenstein M, Lobe C, Lu X, Nagy A, Burke RD, Koop BF, Howard PL (2008) ARS2 is a conserved eukaryotic gene essential for early mammalian development. Mol Cell Biol 28(5):1503–1514. doi:10.1128/MCB.01565-07
Wu Y, Xu Y, Shen Y, Wang C, Guo G, Hu T (2012) Tetramethylpyrazine potentiates arsenic trioxide activity against HL-60 cell lines. Braz J Med Biol Res 45(3):187–196
Yu M, Xue J, Li Y, Zhang W, Ma D, Liu L, Zhang Z (2013) Resveratrol protects against arsenic trioxide-induced nephrotoxicity by facilitating arsenic metabolism and decreasing oxidative stress. Arch Toxicol 87(6):1025–1035. doi:10.1007/s00204-013-1026-4
Zhang J, Ohta T, Maruyama A, Hosoya T, Nishikawa K, Maher JM, Shibahara S, Itoh K, Yamamoto M (2006) BRG1 interacts with Nrf2 to selectively mediate HO-1 induction in response to oxidative stress. Mol Cell Biol 26(21):7942–7952. doi:10.1128/MCB.00700-06
Zhang X, Su Y, Zhang M, Sun Z (2012) Opposite effects of arsenic trioxide on the Nrf2 pathway in oral squamous cell carcinoma in vitro and in vivo. Cancer Lett 318(1):93–98. doi:10.1016/j.canlet.2011.12.005
Zheng LY, Umans JG, Tellez-Plaza M, Yeh F, Francesconi KA, Goessler W, Silbergeld EK, Guallar E, Howard BV, Weaver VM, Navas-Acien A (2013) Urine arsenic and prevalent albuminuria: evidence from a population-based study. Am J Kidney Dis 61(3):385–394. doi:10.1053/j.ajkd.2012.09.011
Zheng L, Kuo CC, Fadrowski J, Agnew J, Weaver VM, Navas-Acien A (2014) Arsenic and chronic kidney disease: a systematic review. Curr Environ Health Rep 1(3):192–207. doi:10.1007/s40572-014-0024-x
Acknowledgments
The authors would like to thank Drs. Preety Sharma, Erik Young, Bo Zhang, Gangkun Lin, Wupeng Liao, Hongning Zhou and Winsome Walker for advice and helpful discussion. This work was supported by Environmental Center Grant P30 ES009089, NIH Grant P01 CA049062 and National Natural Science Foundation of China Grant 81573936 and 81373614.
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Gong, X., Ivanov, V.N. & Hei, T.K. 2,3,5,6-Tetramethylpyrazine (TMP) down-regulated arsenic-induced heme oxygenase-1 and ARS2 expression by inhibiting Nrf2, NF-κB, AP-1 and MAPK pathways in human proximal tubular cells. Arch Toxicol 90, 2187–2200 (2016). https://doi.org/10.1007/s00204-015-1600-z
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DOI: https://doi.org/10.1007/s00204-015-1600-z