This study aimed to investigate the role of purple tomato anthocyanin (PTA) in autophagy induced by chromium(VI) in a chicken hepatocellular carcinoma cell line (LMH cells). LMH cells were exposed to Cr(VI), PTA, and Cr(VI) + PTA. The changes in endoplasmic reticulum (ER) stress, autophagy, related proteins, and COX-2 were detected. Results showed that the cell viability was reduced after Cr(VI) treatment, and the decrease was also restrained by 3-MA or PTA. Levels of ER stress-related proteins (GRP78/Bip and PERK) and COX-2 increased after Cr(VI) treatment, which resulted in an increase in autophagy-related proteins (Beclin1 and LC3-II), inhibition of autophagy pathway protein mTOR, and degradation of autophagy-related protein p62, leading to excessive autophagy and cell damage. Meanwhile, the changes of these indicators induced by Cr(VI) were alleviated by PTA. In conclusion, our study suggested that Cr(VI) can induce excessive autophagy in LMH cells, while PTA can ameliorate Cr(VI)-induced autophagy by inhibiting ER stress.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Chen H, Cao J, Li L, Wu X, Bi R, Klerks PL, Xie L (2016) Maternal transfer and reproductive effects of Cr(VI) in Japanese medaka (Oryzias latipes) under acute and chronic exposures. Aquat Toxicol 171:59–68. https://doi.org/10.1016/j.aquatox.2015.12.011
Chen P, Zhu Y, Wan H, Wang Y, Hao P, Cheng Z, Liu Y, Liu J (2017) Effects of the oral administration of K2Cr2O7 and Na2SeO3 on Ca, Mg, Mn, Fe, Cu, and Zn contents in the heart, liver, spleen, and kidney of chickens. Biol Trace Elem Res 180(2):285–296. https://doi.org/10.1007/s12011-017-0999-x
Zhu Y, Chen P, Wan H, Wang Y, Hao P, Liu Y, Liu J (2018) Selenium-chromium(VI) interaction regulates the contents and correlations of trace elements in chicken brain and serum. Biol Trace Elem Res 181(1):154–163. https://doi.org/10.1007/s12011-017-1038-7
Wang Y, Liu Y, Wan H, Zhu Y, Chen P, Hao P, Cheng Z, Liu J (2017) Moderate selenium dosing inhibited chromium (VI) toxicity in chicken liver. J Biochem Mol Toxicol 31(8). https://doi.org/10.1002/jbt.21916
das Neves RP, Santos TM, De Lourdes Pereira M, de Jesus JP (2016) Comparative histological studies on liver of mice exposed to Cr(VI) and Cr(V) compounds. Hum Exp Toxicol 21(7):365–369. https://doi.org/10.1191/0960327102ht243oa
Xie Y, Xiao F, Luo L, Zhong C (2014) Activation of autophagy protects against ROS-mediated mitochondria-dependent apoptosis in L-02 hepatocytes induced by Cr(VI). Cell Physiol Biochem: international journal of experimental cellular physiology 33(3):705–716. https://doi.org/10.1159/000358646
Liang Q, Xiao Y, Liu K, Zhong C, Zeng M, Xiao F (2018) Cr(VI)-induced autophagy protects L-02 hepatocytes from apoptosis through the ROS-AKT-mTOR pathway. Cell Physiol Biochem 51(4):1863–1878. https://doi.org/10.1159/000495713
Chen P, Geng N, Zhou D, Zhu Y, Xu Y, Liu K, Liu Y, Liu J (2019) The regulatory role of COX-2 in the interaction between Cr(VI)-induced endoplasmic reticulum stress and autophagy in DF-1 cells. Ecotoxicol Environ Saf 170:112–119
Kawaguchi T, Nomura K, Hirayama Y, Kitagawa T (1987) Establishment and characterization of a chicken hepatocellular carcinoma cell line. LMH Cancer Res 47(16):4460–4464
Gabis KK, Gildemeister OS, Pepe JA, Lambrecht RW, Bonkovsky HL (1996) Induction of heme oxygenase-1 in LMH cells. Comparison of LMH cells to primary cultures of chick embryo liver cells. Biochim Biophys Acta Gen Subj 1290(1):113–120. https://doi.org/10.1016/0304-4165(96)00009-8
Xia X, Ling W, Ma J, Xia M, Hou M, Wang Q, Zhu H, Tang Z (2006) An anthocyanin-rich extract from black rice enhances atherosclerotic plaque stabilization in apolipoprotein E-deficient mice. J Nutr 136(8):6. https://doi.org/10.1093/jn/136.8.2220
Guerra MC, Galvano F, Bonsi L, Speroni E, Costa S, Renzulli C, Cervellati R (2007) Cyanidin-3-O-β-glucopyranoside, a natural free-radical scavenger against aflatoxin B1- and ochratoxin A-induced cell damage in a human hepatoma cell line (Hep G2) and a human colonic adenocarcinoma cell line (CaCo-2). Br J Nutr 94(02):211–220. https://doi.org/10.1079/bjn20051425
Chun OK, Kim DO, Lee CY (2003) Superoxide radical scavenging activity of the major polyphenols in fresh plums. J Agric Food Chem 51(27):6. https://doi.org/10.1021/jf034740d
Galvano F, La Fauci L, Lazzarino G, Fogliano V, Ritieni A, Ciappellano S, Battistini NC, Tavazzi B, Galvano G (2004) Cyanidins: metabolism and biological properties. J Nutr Biochem 15(1):10. https://doi.org/10.1016/s0955-2863(03)00133-5
Chanoca A, Kovinich N, Burkel B, Stecha S, Bohorquez-Restrepo A, Ueda T, Eliceiri KW, Grotewold E, Otegui MS (2015) Anthocyanin vacuolar inclusions form by a microautophagy mechanism. Plant Cell 27(9):2545–2559. https://doi.org/10.1105/tpc.15.00589
Zhang B, Buya M, Qin W, Sun C, Cai H, Xie Q, Xu B, Wu Y (2013) Anthocyanins from Chinese bayberry extract activate transcription factor Nrf2 in β cells and negatively regulate oxidative stress-induced autophagy. J Agric Food Chem 61(37):8765–8772. https://doi.org/10.1021/jf4012399
Xie N, Geng N, Zhou D, Xu Y, Liu K, Liu Y, Liu J (2018) Protective effects of anthocyanin against apoptosis and oxidative stress induced by arsanilic acid in DF-1 cells. Mol Biol Rep 46:301–308. https://doi.org/10.1007/s11033-018-4472-5
Zhang B, Buya M, Qin W, Sun C, Cai H, Xie Q, Xu B, Wu Y (2013) Anthocyanins from Chinese bayberry extract activate transcription factor Nrf2 in beta cells and negatively regulate oxidative stress-induced autophagy. J Agric Food Chem 61(37):8765–8772. https://doi.org/10.1021/jf4012399
Huang J, Klionsky DJ (2007) Autophagy and human disease. Cell Cycle 6(15):1837–1849
Cuervo AM (2004) Autophagy: in sickness and in health. Trends Cell Biol 14(2):70–77. https://doi.org/10.1016/j.tcb.2003.12.002
Yang F, Liao J, Pei R, Yu W, Han Q, Li Y, Guo J, Hu L, Pan J, Tang Z (2018) Autophagy attenuates copper-induced mitochondrial dysfunction by regulating oxidative stress in chicken hepatocytes. Chemosphere 204:36–43. https://doi.org/10.1016/j.chemosphere.2018.03.192
Wang Z, Li S, Ren R, Li J, Cui X (2015) Recombinant buckwheat trypsin inhibitor induces mitophagy by directly targeting mitochondria and causes mitochondrial dysfunction in Hep G2 cells. J Agric Food Chem 63(35):7795–7804. https://doi.org/10.1021/acs.jafc.5b02644
Lee YH, Su SB, Huang CC, Sheu HM, Tsai JC, Lin CH, Wang YJ, Wang BJ (2014) N-Acetylcysteine attenuates hexavalent chromium-induced hypersensitivity through inhibition of cell death, ROS-related signaling and cytokine expression. PLoS One 9(9):e108317
Wang SH, Shih YL, Ko WC, Wei YH, Shih CM (2008) Cadmium-induced autophagy and apoptosis are mediated by a calcium signaling pathway. Cell Mol Life Sci 65(22):3640–3652. https://doi.org/10.1007/s00018-008-8383-9
Hussain S, Al-Nsour F, Rice AB, Marshburn J, Yingling B, Ji Z, Zink JI, Walker NJ, Garantziotis S (2012) Cerium dioxide nanoparticles induce apoptosis and autophagy in human peripheral blood monocytes. ACS Nano 6(7):5820–5829. https://doi.org/10.1021/nn302235u
Sui X, Chen R, Wang Z, Huang Z, Kong N, Zhang M, Han W, Lou F, Yang J, Zhang Q, Wang X, He C, Pan H (2013) Autophagy and chemotherapy resistance: a promising therapeutic target for cancer treatment. Cell Death Dis 4:e838. https://doi.org/10.1038/cddis.2013.350
Dai J, Ji Y, Wang W, Kim D, Fai LY, Wang L, Luo J, Zhang Z (2017) Loss of fructose-1,6-bisphosphatase induces glycolysis and promotes apoptosis resistance of cancer stem-like cells: an important role in hexavalent chromium-induced carcinogenesis. Toxicology & Applied Pharmacology 331:164–173. https://doi.org/10.1016/j.taap.2017.06.014
Matthias J, Messling S, Eichinger L (2016) The two Dictyostelium autophagy eight proteins, ATG8a and ATG8b, associate with the autophagosome in succession. Eur J Cell Biol 95(1):15–25. https://doi.org/10.1016/j.ejcb.2015.10.007
Clifford M (2004) Diet-derived phenols in plasma and tissues and their implications for health. Planta Med 70(12):1103–1114. https://doi.org/10.1055/s-2004-835835
Hitoshi M, Yuko N, Shuji T, Satoru K, Masao H (2003) Stimulatory effect of cyanidin 3-glycosides on the regeneration of rhodopsin. J Agric Food Chem 51(12):3560–3563. https://doi.org/10.1128/IAI.01773-14
Wang H, Nair MG, Strasburg GM, Chang YC, Booren AM, Gray JI, Dewitt DL (1999) Antioxidant and antiinflammatory activities of anthocyanins and their aglycon, cyanidin, from tart cherries. J Nat Prod 62(2):294–296. https://doi.org/10.1021/np980501m
Xingchen Z, Feng S, Jianli Z, Rutao L (2011) Composition and stability of anthocyanins from purple Solanum tuberosum and their protective influence on Cr(VI) targeted to bovine serum albumin. J Agric Food Chem 59(14):7902–7909
Klionsky D, Abdelmohsen K, Abe A, Abedin M, Abeliovich A, AA A, Adachi H et al (2016) Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy 12(1):222. https://doi.org/10.1080/15548627.2015.1100356
Fang CY, Chen JS, Chang SK, Shen CH (2018) Reversine induces autophagic cell death through the AMP-activated protein kinase pathway in urothelial carcinoma cells. Anti-Cancer Drugs 29(1):29–39. https://doi.org/10.1097/CAD.0000000000000563
Zhang Y, Xiao F, Liu X, Liu K, Zhou X, Zhong C (2017) Cr(VI) induces cytotoxicity in vitro through activation of ROS-mediated endoplasmic reticulum stress and mitochondrial dysfunction via the PI3K/Akt signaling pathway. Toxicol in Vitro 41:232–244. https://doi.org/10.1016/j.tiv.2017.03.003
Wang Z, Yin F, Xu J, Zhang T, Wang G, Mao M, Wang Z, Sun W, Han J, Yang M, Jiang Y, Hua Y, Cai Z (2019) CYT997(Lexibulin) induces apoptosis and autophagy through the activation of mutually reinforced ER stress and ROS in osteosarcoma. J Exp Clin Cancer Res 38(1):44. https://doi.org/10.1186/s13046-019-1047-9
Fouillet A, Levet C, Virgone A, Robin M, Dourlen P, Rieusset J, Belaidi E, Ovize M, Touret M, Nataf S, Mollereau B (2014) ER stress inhibits neuronal death by promoting autophagy. Autophagy 8(6):915–926. https://doi.org/10.4161/auto.19716
Periyasamy P, Guo ML, Buch S (2016) Cocaine induces astrocytosis through ER stress-mediated activation of autophagy. Autophagy 12(8):1310–1329. https://doi.org/10.1080/15548627.2016.1183844
Louessard M, Bardou I, Lemarchand E, Thiebaut AM, Parcq J, Leprince J, Terrisse A, Carraro V, Fafournoux P, Bruhat A, Orset C, Vivien D, Ali C, Roussel BD (2017) Activation of cell surface GRP78 decreases endoplasmic reticulum stress and neuronal death. Cell Death Differ 24(9):1518–1529. https://doi.org/10.1038/cdd.2017.35
Cubillos-Ruiz JR, Bettigole SE, Glimcher LH (2017) Tumorigenic and immunosuppressive effects of endoplasmic reticulum stress in cancer. Cell 168(4):692–706. https://doi.org/10.1016/j.cell.2016.12.004
Walter P, Ron D (2011) The unfolded protein response: from stress pathway to homeostatic regulation. Science 334(6059):1081–1086. https://doi.org/10.1126/science.1209038
Wargasetia T, Shahib N, Martaadisoebrata D, Dhianawaty D, Hernowo B (2015) Characterization of apoptosis and autophagy through Bcl-2 and Beclin-1 immuno expression in gestational trophoblastic disease. Iran J Reprod Med 13(7):8. https://doi.org/10.1128/IAI.01773-14
Yang Z, Wilkie-Grantham RP, Yanagi T, Shu C-W, Matsuzawa S-i, Reed JC (2015) ATG4B (autophagin-1) phosphorylation modulates autophagy. J Biol Chem 290(44):26549–26561. https://doi.org/10.1074/jbc.M115.658088
Li H, Song Y, He Z, Chen X, Wu X, Li X, Bai X, Liu W, Li B, Wang S, Han Y, Xu L, Zhang D, Li J, Chai R, Wang H, Fan Z (2018) Meclofenamic acid reduces reactive oxygen species accumulation and apoptosis, inhibits excessive autophagy, and protects hair cell-like HEI-OC1 cells from cisplatin-induced damage. Front Cell Neurosci 12. https://doi.org/10.3389/fncel.2018.00139
Ryter SW, Mizumura K, Choi AMK (2014) The impact of autophagy on cell death modalities. Int J Cell Biol 2014:1–12. https://doi.org/10.1155/2014/502676
Czaja MJ, Ding W-X, Donohue TM, Friedman SL, Kim J-S, Komatsu M, Lemasters JJ, Lemoine A, Lin JD, Ou JH, Perlmutter DH, Randall G, Ray RB, Tsung A, Yin X-M (2014) Functions of autophagy in normal and diseased liver. Autophagy 9(8):1131–1158. https://doi.org/10.4161/auto.25063
The project was supported by the National Nature Science Foundation of China (No. 31872535), National Key R&D Program (2016YFD0501208), Shandong Natural Science Foundation of China (ZR2018MC027, ZR2016CQ29), and Funds of Shandong “Double Tops” Program. PTA was donated by Prof. Ding Xinhua of Shandong Agricultural University.
Conflict of Interest
The authors declare that they have no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Liu, K., Chen, P., Lu, J. et al. Protective Effect of Purple Tomato Anthocyanidin on Chromium(VI)-Induced Autophagy in LMH Cells by Inhibiting Endoplasmic Reticulum Stress. Biol Trace Elem Res 194, 570–580 (2020). https://doi.org/10.1007/s12011-019-01795-3
- Chromium (VI)
- ER stress
- LMH cells