Acteoside from Conandron ramondioides Reduces Microcystin-LR Cytotoxicity by Inhibiting Intracellular Uptake Mediated by OATP1B3

Abstract

The hepatotoxin microcystin-LR is a strong inhibitor of serine/threonine protein phosphatase (PP) 1 and PP2A. The onset of its cytotoxicity depends on its selective uptake via the hepatocyte uptake transporters, organic anion transporting polypeptide (OATP) 1B1 and OATP1B3. Understanding and preventing the cytotoxicity of microcystin-LR is crucial to maintain human health. This chemoprevention study demonstrates that the herbal plant extract of iwajisha (20 µg/mL) reduced microcystin-LR cytotoxicity in OATP1B3-expressing cells by approximately six times. In addition, 20 µM acteoside, which is one of the major compounds in iwajisha, reduced microcystin-LR cytotoxicity by approximately 7.4 times. Acteoside could also reduce the cytotoxicity of other compounds, such as okadaic acid and nodularin, which are both substrates of OATP1B3 and inhibitors of PP1/PP2A. To investigate the mechanism by which the cytotoxicity of microcystin-LR is attenuated by acteosides, microcystin-LR and microcystin-LR-binding proteins in cells were examined after microcystin-LR and acteosides were co-exposed. Thus, acteoside noncompetitively inhibited microcystin-LR uptake by OATP1B3-expressing cells. Furthermore, acteoside inhibited the intracellular interaction of microcystin-LR with its binding protein(s), including the 22 kDa protein. Furthermore, using immunoblot analysis, acteoside induced the phosphorylation of extracellular signal-regulated kinase (ERK), which is one of the survival signaling molecules. These results suggest that acteoside reduces microcystin-LR cytotoxicity through several mechanisms, including the inhibition of microcystin-LR uptake via OATP1B3, and decreased interaction between microcystin-LR and its binding protein(s), and that ERK signaling activation contributes to the attenuation effect of acteoside against microcystin-LR cytotoxicity.

Key words

Conandron ramondioides - Gesneriaceae - iwajisha - acteoside - microcystin-LR - OATP1B3

Publication History

Received: 27 April 2022

Accepted after revision: 28 October 2022

Article published online:
10 January 2023

• References

• 1 van Apeldoorn ME, van Egmond HP, Speijers GJ, Bakker GJ. Toxins of cyanobacteria. Mol Nutr Food Res 2007; 51: 7-60
• 2 Gehringer MM. Microcystin-LR and okadaic acid-induced cellular effects: A dualistic response. FEBS Lett 2004; 557: 1-8
• 3 Komatsu M, Furukawa T, Ikeda R, Takumi S, Nong Q, Aoyama K, Akiyama S, Keppler D, Takeuchi T. Involvement of mitogen-activated protein kinase signaling pathways in microcystin-LR-induced apoptosis after its selective uptake mediated by OATP1B1 and OATP1B3. Toxicol Sci 2007; 97: 407-416
• 4 Takumi S, Komatsu M, Furukawa T, Ikeda R, Sumizawa T, Akenaga H, Maeda Y, Aoyama K, Arizono K, Ando S, Takeuchi T. p53 plays an important role in cell fate determination after exposure to Microcystin-LR. Environ Health Perspect 2010; 118: 1292-1298
• 5 Takumi S, Ikema S, Hanyu T, Shima Y, Kurimoto T, Shiozaki K, Sugiyama Y, Park HD, Ando S, Furukawa T, Komatsu M. Naringin attenuates the cytotoxicity of hepatotoxin microcystin-LR by the curious mechanisms to OATP1B1- and OATP1B3-expressing cells. Environ Toxicol Pharmacol 2015; 39: 974-981
• 6 Herfindal L, Myhren L, Kleppe R, Krakstad C, Selheim F, Jokela J, Sivonen K, Døskeland SO. Nostocyclopeptide-M1: A potent, nontoxic inhibitor of the hepatocyte drug transporters OATP1B3 and OATP1B1. Mol Pharm 2011; 8: 360-367
• 7 Ikema S, Takumi S, Maeda Y, Kurimoto T, Bohda S, Chigwechokha PK, Sugiyama Y, Shiozaki K, Furukawa T, Komatsu M. Okadaic acid is taken-up into the cells mediated by human hepatocytes transporter OATP1B3. Food Chem Toxicol 2015; 83: 229-236
• 8 Letschert K, Faulstich H, Keller D. Molecular characterization and inhibition of amanitin uptake into human hepatocytes. Toxicol Sci 2006; 91: 140-149
• 9 Fehrenbach T, Cui Y, Faulstich H, Keppler D. Characterization of the transport of the bicyclic peptide phalloidin by human hepatic transport proteins. Naunyn Schmiedebergs Arch Pharmacol 2003; 368: 415-420
• 10 Miyaichi Y, Ohichi M, Yaguchi K, Kawata Y, Kizu H. Studies on the constituents of the leaves of Conandron ramondioides. J Nat Med 2006; 60: 159-160
• 11 Dai X, Su S, Cai H, Wei D, Zheng T, Zhu Z, Yan H, Shang E, Guo S, Qian D, Duan J. Comparative pharmacokinetics of acteoside from total glycoside extracted from leaves of Rehmannia and Dihuangye total glycoside capsule in normal and diabetic nephropathy rats. Biomed Chromatogr 2017; 31: e4013
• 12 Xiang Q, Kadota S, Tani T, Namba T. Antioxidative effects of phenylethanoids from Cistanche deserticola. Biol Pharm Bull 1996; 19: 1580-1585
• 13 Chathuranga K, Kim MS, Lee H, Kim T, Kim J, Chathuranga WAG, Ekanayaka P, Wijerathne HMSM, Cho W, Kim HI, Ma JY, Lee J. Anti-respiratory syncytial virus activity of Plantago asiatica and Clerodendrum trichotomum extracts in vitro and in vivo . Viruses 2019; 11: 604
• 14 Nonaka G, Nishioka I. Bitter phenylpropanoid glycosides from Conandron ramoidioides . Phytochemistry 1977; 16: 1265-1267
• 15 Xiufen W, Hiramatsu N, Matsubara M. The antioxidative activity of traditional Japanese herbs. Biofactors 2004; 21: 281-284
• 16 Lee KJ, Woo E, Choi CY, Shin DW, Lee DG, You HJ, Jeong HG. Protecive effect of acteoside on carbon tetrachloride-induced hepatotoxicity. Life Sci 2004; 74: 1051-1064
• 17 Nong Q, Komatsu M, Izumo K, Indo HP, Xu B, Aoyama K, Majima HJ, Horiuchi M, Morimoto K, Takeuchi T. Involvement of reactive oxygen species in microcystin-LR-induced cytogenotoxicity. Free Radic Res 2007; 41: 1326-1337
• 18 Gonzalez FJ. Role of cytochrome P450 in chemical toxicity and oxidative stress: Studies with CYP2E1. Mutat Res 2005; 569: 101-110
• 19 Hu Y, Da L. Insights into the selective binding and toxic mechanism of microcystin to catalase. Spectrochim Acta A Mol Biomol Spectrosc 2014; 121: 230-237
• 20 Mori T, Kubo T, Kaya K, Hosoya K. Comprehensive study of proteins that interact with microcystin-LR. Anal Bioanal Chem 2012; 402: 1137-1147
• 21 Chen T, Cui J, Liang Y, Xin X, Young DO, Chen C, Shen P. Identification of human liver mitochondrial aldehyde dehydrogenase as a potential target for microcystin-LR. Toxicology 2006; 220: 71-80
• 22 Mikhailov A, Härmälä-Braskén A, Hellman J, Meriluoto J, Eriksson JE. Identification of ATP-synthase as a novel intracellular target for microcystin-LR. Chem Biol Interact 2003; 142: 223-237
• 23 Zhu Z, Zhang L, Shi G. Proteasome as a molecular target of Microcystin-LR. Toxins (Basel) 2015; 7: 2221-2231
• 24 Takumi S, Shimono T, Ikema S, Hotta Y, Chigwechokha PK, Shiozaki K, Sugiyama Y, Hashimoto M, Furukawa T, Komatsu M. Overexpression of carboxylesterase contributes to the attenuation of cyanotoxin microcystin-LR toxicity. Comp Biochem Physiol C Toxicol Pharmacol 2017; 194: 22-27
• 25 Son Y, Lee S, Kim S, Jang Y, Chun J, Lee J. Acteoside inhibits melanogenesis in B16F10 cells through ERK activation and tyrosinase down-regulation. J Pharm Pharmacol 2011; 63: 1309-1319
• 26 König J, Cui Y, Nies AT, Keppler D. A novel human organic anion transporting polypeptide localized to the basolateral hepatocyte membrane. Am J Physiol Gastrointest Liver Physiol 2000; 278: 156-164
• 27 Komatsu M, Sumizawa T, Mutoh M, Chen Z, Terada K, Furukawa T, Yang X, Gao H, Miura N, Sugiyama T, Akiyama S. Copper-transporting P-type adenosine triphosphatase (ATP7B) is associated with cisplatin resistance. Cancer Res 2000; 60: 1312-1316