Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
권호 표지
DOI QR코드

DOI QR Code

Enhancing effect of Panax ginseng on Zip4-mediated zinc influx into the cytosol

  • Ikeda, Yoshito (Laboratory of Medicinal Cell Biology, Kobe Pharmaceutical University) ;
  • Munekane, Masayuki (Laboratory of Biophysical Chemistry, Kobe Pharmaceutical University) ;
  • Yamada, Yasuyuki (Laboratory of Medicinal Cell Biology, Kobe Pharmaceutical University) ;
  • Kawakami, Mizuki (Laboratory of Medicinal Cell Biology, Kobe Pharmaceutical University) ;
  • Amano, Ikuko (Laboratory of Medicinal Cell Biology, Kobe Pharmaceutical University) ;
  • Sano, Kohei (Laboratory of Biophysical Chemistry, Kobe Pharmaceutical University) ;
  • Mukai, Takahiro (Laboratory of Biophysical Chemistry, Kobe Pharmaceutical University) ;
  • Kambe, Taiho (Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University) ;
  • Shitan, Nobukazu (Laboratory of Medicinal Cell Biology, Kobe Pharmaceutical University)
  • Received : 2020.10.28
  • Accepted : 2021.06.09
  • Published : 2022.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

Background: Zinc homeostasis is essential for human health and is regulated by several zinc transporters including ZIP and ZnT. ZIP4 is expressed in the small intestine and is important for zinc absorption from the diet. We investigated in the present study the effects of Panax ginseng (P. ginseng) extract on modulating Zip4 expression and cellular zinc levels in mouse Hepa cells. Methods: Hepa cells were transfected with a luciferase reporter plasmid that contains metal-responsive elements, incubated with P. ginseng extract, and luciferase activity was measured. Using 65ZnCl2, zinc uptake in P. ginseng-treated cells was measured. The expression of Zip4 mRNA and protein in Hepa cells was also investigated. Finally, using a luciferase reporter assay system, the effects of several ginsenosides were monitored. Results: The luciferase activity in cells incubated with P. ginseng extract was significantly higher than that of control cells cultured in normal medium. Hepa cells treated with P. ginseng extract exhibited higher zinc uptake. P. ginseng extract induced Zip4 mRNA expression, which resulted in an enhancement of Zip4 protein expression. Furthermore, some ginsenosides, such as ginsenoside Rc and Re, enhanced luciferase activity driven by intracellular zinc levels. Conclusion: P. ginseng extract induced Zip4 expression at the mRNA and protein level and resulted in higher zinc uptake in Hepa cells. Some ginsenosides facilitated zinc influx. On the basis of these results, we suggest a novel effect of P. ginseng on Zip4-mediated zinc influx, which may provide a new strategy for preventing zinc deficiency.

Keywords

Acknowledgement

We thank Dr. Yumi Nishiyama and Yudai Hamaguchi (Kobe Pharmaceutical University, Japan) for help with the experiments. We thank Dr. Glen K. Andrews (University of Kansas Medical Center) for providing Hepa cells. The authors would like to thank Enago (www.enago.jp) for the English language review.

References

  1. Kochanczyk T, Drozd A, Krezel A. Relationship between the architecture of zinc coordination and zinc binding affinity in proteins-insights into zinc regulation. Metallomics 2015;7:244-57. https://doi.org/10.1039/C4MT00094C
  2. Kambe T, Tsuji T, Hashimoto A, Itsumura N. The physiological, biochemical, and molecular roles of zinc transporters in zinc homeostasis and metabolism. Physiol Rev 2015;95:749-84. https://doi.org/10.1152/physrev.00035.2014
  3. Prasad AS. Discovery of human zinc deficiency and studies in an experimental human model. Am J Clin Nutr 1991;53:403-12. https://doi.org/10.1093/ajcn/53.2.403
  4. Van Wouwe JP. Clinical and laboratory assessment of zinc deficiency in Dutch children. A review. Biol Trace Elem Res 1995;49:211-25. https://doi.org/10.1007/BF02788969
  5. Goto T, Komai M, Suzuki H, Furukawa Y. Long-term zinc deficiency decreases taste sensitivity in rats. J Nutr 2001;131:305-10. https://doi.org/10.1093/jn/131.2.305
  6. Hojyo S, Fukada T. Roles of zinc signaling in the immune system. J Immunol Res 2016;2016:6762343. https://doi.org/10.1155/2016/6762343
  7. Dufner-Beattie J, Wang F, Kuo YM, Gitschier J, Eide D, Andrews GK. The acrodermatitis enteropathica gene ZIP4 encodes a tissue-specific, zinc-regulated zinc transporter in mice. J Biol Chem 2003;278:33474-81. https://doi.org/10.1074/jbc.M305000200
  8. Maverakis E, Fung MA, Lynch PJ, Draznin M, Michael DJ, Ruben B, Fazel N. Acrodermatitis enteropathica and an overview of zinc metabolism. J Am Acad Dermatol 2007;56:116-24. https://doi.org/10.1016/j.jaad.2006.08.015
  9. Dufner-Beattie J, Kuo YM, Gitschier J, Andrews GK. The adaptive response to dietary zinc in mice involves the differential cellular localization and zinc regulation of the zinc transporters ZIP4 and ZIP5. J Biol Chem 2004;279:49082-90. https://doi.org/10.1074/jbc.M409962200
  10. Liuzzi JP, Bobo JA, Lichten LA, Samuelson DA, Cousins RJ. Responsive transporter genes within the murine intestinal-pancreatic axis form a basis of zinc homeostasis. Proc Natl Acad Sci U S A 2004;101:14355-60. https://doi.org/10.1073/pnas.0406216101
  11. Weaver BP, Dufner-Beattie J, Kambe T, Andrews GK. Novel zinc-responsive post-transcriptional mechanisms reciprocally regulate expression of the mouse Slc39a4 and Slc39a5 zinc transporters (Zip4 and Zip5). Biol Chem 2007;388:1301-12. https://doi.org/10.1515/BC.2007.149
  12. Hashimoto A, Ohkura K, Takahashi M, Kizu K, Narita H, Enomoto S, Miyamae Y, Masuda S, Nagao M, Irie K, et al. Soybean extracts increase cell surface ZIP4 abundance and cellular zinc levels: a potential novel strategy to enhance zinc absorption by ZIP4 targeting. Biochem J 2015;472:183-93. https://doi.org/10.1042/BJ20150862
  13. Kambe T, Andrews GK. Novel proteolytic processing of the ectodomain of the zinc transporter ZIP4 (SLC39A4) during zinc deficiency is inhibited by acrodermatitis enteropathica mutations. Mol Cell Biol 2009;29:129-39. https://doi.org/10.1128/MCB.00963-08
  14. Sandstead HH, Henriksen LK, Greger JL, Prasad AS, Good RA. Zinc nutriture in the elderly in relation to taste acuity, immune response, and wound healing. Am J Clin Nutr 1982;36:1046-59. https://doi.org/10.1093/ajcn/36.5.1046
  15. Haase H, Mocchegiani E, Rink L. Correlation between zinc status and immune function in the elderly. Biogerontology 2006;7:421-8. https://doi.org/10.1007/s10522-006-9057-3
  16. Kogirima M, Kurasawa R, Kubori S, Sarukura N, Nakamori M, Okada S, Kamioka H, Yamamoto S. Ratio of low serum zinc levels in elderly Japanese people living in the central part of Japan. Eur J Clin Nutr 2007;61:375-81. https://doi.org/10.1038/sj.ejcn.1602520
  17. Mancuso C, Santangelo R. Panax ginseng and Panax quinquefolius: from pharmacology to toxicology. Food Chem Toxicol 2017;107:362-72. https://doi.org/10.1016/j.fct.2017.07.019
  18. Shibata S. Chemistry and cancer preventing activities of ginseng saponins and some related triterpenoid compounds. J Korean Med Sci 2001;16(Suppl):S28-37. https://doi.org/10.3346/jkms.2001.16.S.S28
  19. Ru W, Wang D, Xu Y, He X, Sun YE, Qian L, Zhou X, Qin Y. Chemical constituents and bioactivities of Panax ginseng (C. A. Mey.). Drug Discov Ther 2015;9:23-32. https://doi.org/10.5582/ddt.2015.01004
  20. Huang Q, Wang T, Wang HY. Ginsenoside Rb2 enhances the anti-inflammatory effect of omega-3 fatty acid in LPS-stimulated RAW264.7 macrophages by upregulating GPR120 expression. Acta Pharmacol Sin 2017;38:192-200. https://doi.org/10.1038/aps.2016.135
  21. Yu T, Yang Y, Kwak YS, Song GG, Kim MY, Rhee MH, Cho JY. Ginsenoside Rc from Panax ginseng exerts anti-inflammatory activity by targeting TANK-binding kinase 1/interferon regulatory factor-3 and p38/ATF-2. J Ginseng Res 2017;41:127-33. https://doi.org/10.1016/j.jgr.2016.02.001
  22. Kim JH. Cardiovascular diseases and Panax ginseng: a review on molecular mechanisms and medical applications. J Ginseng Res 2012;36:16-26. https://doi.org/10.5142/jgr.2012.36.1.16
  23. Yang JW, Kim SS. Ginsenoside Rc promotes anti-adipogenic activity on 3T3-L1 adipocytes by down-regulating C/EBPalpha and PPARgamma. Molecules 2015;20:1293-303. https://doi.org/10.3390/molecules20011293
  24. Sun M, Ye Y, Xiao L, Duan X, Zhang Y, Zhang H. Anticancer effects of ginsenoside Rg3 (review). Int J Mol Med 2017;39:507-18. https://doi.org/10.3892/ijmm.2017.2857
  25. Matsuura H. Saponins in garlic as modifiers of the risk of cardiovascular disease. J Nutr 2001;131. 1000S-5S. https://doi.org/10.1093/jn/131.3.1000S
  26. Liu MJ, Wang Z, Ju Y, Wong RN, Wu QY. Diosgenin induces cell cycle arrest and apoptosis in human leukemia K562 cells with the disruption of Ca2+ homeostasis. Cancer Chemother Pharmacol 2005;55:79-90. https://doi.org/10.1007/s00280-004-0849-3
  27. Radtke F, Heuchel R, Georgiev O, Hergersberg M, Gariglio M, Dembic Z, Schaffner W. Cloned transcription factor MTF-1 activates the mouse metallothionein I promoter. EMBO J 1993;12:1355-62. https://doi.org/10.1002/j.1460-2075.1993.tb05780.x
  28. Ikeda Y, Tsuchiya H, Hama S, Kajimoto K, Kogure K. Resistin regulates the expression of plasminogen activator inhibitor-1 in 3T3-L1 adipocytes. Biochem Biophys Res Commun 2014;448:129-33. https://doi.org/10.1016/j.bbrc.2014.03.076
  29. Hashimoto A, Nakagawa M, Tsujimura N, Miyazaki S, Kizu K, Goto T, Komatsu Y, Matsunaga A, Shirakawa H, Narita H, et al. Properties of Zip4 accumulation during zinc deficiency and its usefulness to evaluate zinc status: a study of the effects of zinc deficiency during lactation. Am J Physiol Regul Integr Comp Physiol 2016;310:R459-68. https://doi.org/10.1152/ajpregu.00439.2015
  30. Liuzzi JP, Guo L, Chang SM, Cousins RJ. Kruppel-like factor 4 regulates adaptive expression of the zinc transporter Zip4 in mouse small intestine. Am J Physiol Gastrointest Liver Physiol 2009;296:G517-23. https://doi.org/10.1152/ajpgi.90568.2008
  31. Shields JM, Christy RJ, Yang VW. Identification and characterization of a gene encoding a gut-enriched Kruppel-like factor expressed during growth arrest. J Biol Chem 1996;271:20009-17. https://doi.org/10.1074/jbc.271.33.20009
  32. Kim MO, Kim SH, Cho YY, Nadas J, Jeong CH, Yao K, Kim DJ, Yu DH, Keum YS, Lee KY, et al. ERK1 and ERK2 regulate embryonic stem cell self-renewal through phosphorylation of Klf4. Nat Struct Mol Biol 2012;19:283-90. https://doi.org/10.1038/nsmb.2217
  33. Evans PM, Zhang W, Chen X, Yang J, Bhakat KK, Liu C. Kruppel-like factor 4 is acetylated by p300 and regulates gene transcription via modulation of histone acetylation. J Biol Chem 2007;282:33994-4002. https://doi.org/10.1074/jbc.M701847200
  34. Lim KH, Kim SR, Ramakrishna S, Baek KH. Critical lysine residues of Klf4 required for protein stabilization and degradation. Biochem Biophys Res Commun 2014;443:1206-10. https://doi.org/10.1016/j.bbrc.2013.12.121
  35. Hu D, Gur M, Zhou Z, Gamper A, Hung MC, Fujita N, Lan L, Bahar I, Wan Y. Interplay between arginine methylation and ubiquitylation regulates KLF4-mediated genome stability and carcinogenesis. Nat Commun 2015;6:8419. https://doi.org/10.1038/ncomms9419
  36. Jie Y, He W, Yang X, Chen W. Kruppel-like factor 4 acts as a potential therapeutic target of Sijunzi decoction for treatment of colorectal cancer. Cancer Gene Ther 2017;24:361-6. https://doi.org/10.1038/cgt.2017.25
  37. Zheng CY, Song LL, Wen JK, Li LM, Guo ZW, Zhou PP, Wang C, Li YH, Ma D, Zheng B. Tongxinluo (TXL), a traditional Chinese medicinal compound, improves endothelial function after chronic hypoxia both in vivo and in vitro. J Cardiovasc Pharmacol 2015;65:579-86. https://doi.org/10.1097/FJC.0000000000000226
  38. Peng B, He R, Xu Q, Yang Y, Hu Q, Hou H, Liu X, Li J. Ginsenoside 20(S)-protopanaxadiol inhibits triple-negative breast cancer metastasis in vivo by targeting EGFR-mediated MAPK pathway. Pharmacol Res 2019;142:1-13. https://doi.org/10.1016/j.phrs.2019.02.003
  39. Kim DH, Park CH, Park D, Choi YJ, Park MH, Chung KW, Kim SR, Lee JS, Chung HY. Ginsenoside Rc modulates Akt/FoxO1 pathways and suppresses oxidative stress. Arch Pharm Res 2014;37:813-20. https://doi.org/10.1007/s12272-013-0223-2
  40. Saw CLL, Yang AY, Cheng DC, Boyanapalli SSS, Su ZY, Khor TO, Gao S, Wang J, Jiang ZH, Kong ANT. Pharmacodynamics of ginsenosides: antioxidant activities, activation of Nrf2 and potential synergistic effects of combinations. Chem Res Toxicol 2012;25(8):1574-80. https://doi.org/10.1021/tx2005025
  41. Park SJ, Lee D, Kim D, Lee M, In G, Han ST, Kim SW, Lee MH, Kim OK, Lee J. The non-saponin fraction of Korean Red Ginseng (KGC05P0) decreases glucose uptake and transport in vitro and modulates glucose production via down-regulation of the PI3K/AKT pathway in vivo. J Ginseng Res 2020;44:362-72. https://doi.org/10.1016/j.jgr.2019.12.004
  42. Frank TS, Sun X, Zhang Y, Yang J, Fisher WE, Gingras MC, Li M. Genomic profiling guides the choice of molecular targeted therapy of pancreatic cancer. Cancer Lett 2015;363(1):1-6. https://doi.org/10.1016/j.canlet.2015.04.009