The Extract of Ziziphus jujuba Fruit (Jujube) Induces Expression of Erythropoietin Via Hypoxia-Inducible Factor-1α in Cultured Hep3B Cells

Abstract

The fruit of Ziziphus jujuba Mill., known as jujube or Chinese date, is commonly consumed as health supplement or herbal medicine worldwide. To study the beneficial role of jujube in enhancing hematopoietic function, we investigated its roles on the expression of erythropoietin in cultured Hep3B human hepatocellular carcinoma cells. Application of chemically standardized jujube water extract stimulated erythropoietin expression in a dose-dependent manner, with the highest response by ~ 100 % of increase. A plasmid containing hypoxia response element, a critical regulator for erythropoietin transcription, was transfected into Hep3B cells. Application of jujube water extract onto the transfected cells induced the transcriptional activity of the hypoxia response element. To account for its transcriptional activation, the expression of hypoxia-inducible factor-1α was increased after treatment with jujube water extract: the increase was in both mRNA and protein levels. These results confirmed the hematopoietic function of jujube in the regulation of erythropoietin expression in liver cells.

Key words

Ziziphus jujuba - Rhamnaceae - erythropoietin - hypoxia response element - hypoxia-inducible factor

• References

• 1 Xu YL, Miao MS, Sun YH, Miao YY. Effects of Fructus Jujubae polysaccharide on the hematopoietic function in mice model of both “Qi” and “Blood” deficiencies. Chin J Clin Rehabil 2004; 8: 5050-5051
• 2 Schwenger V, Morath C, Zeier M. Use of erythropoietin after solid organ transplantation. Nephrol Dial Transplant 2007; 22: 47-49
• 3 Jelkmann W. Erythropoietin: structure, control of production, and function. Physiol Rev 1992; 72: 449-489
• 4 Chen JP, Li ZG, Maiwulanjiang M, Zhang WL, Zhan JYX, Lam CTW, Zhu KY, Yao P, Choi RCY, Lau DTW, Dong TT, Tsim KWK. Chemical and biological assessment of Ziziphus jujuba fruits from China: different geographical sources and developmental stages. J Agric Food Chem 2013; 61: 7315-7324
• 5 Chen JP, Maiwulanjiang M, Lam KYC, Zhang WL, Zhan JYX, Lam CTW, Xu SL, Zhu KY, Yao P, Lau DTW, Dong TTX, Tsim KWK. A standardized extract of the fruit of Ziziphus jujuba (jujube) induces neuronal differentiation of cultured PC12 cells: a signaling mediated by protein kinase A. J Agric Food Chem 2014; 62: 1890-1897
• 6 Chen JP, Du CYQ, Lam KYC, Zhang WL, Lam CTW, Yan AL, Yao P, Lau DTW, Dong TTX, Tsim KWK. The standardized extract of Ziziphus jujuba fruit (jujube) regulates pro-inflammatory cytokine expression in cultured murine macrophages: suppression of lipopolysaccharide-stimulated NF-κB activity. Phytother Res 2014; DOI: 10.1002/ptr.516.
• 7 Goldberg MA, Glass GA, Cunningham JM, Bunn HF. The regulated expression of erythropoietin by two human hepatoma cell lines. Proc Natl Acad Sci U S A 1987; 84: 7972-7976
• 8 Gao QT, Cheung JKH, Choi RCY, Cheung AWH, Lv J, Jiang ZY, Duan R, Zhao KJ, Ding AW, Dong TTX, Tsim KWK. A Chinese herbal decoction prepared from Radix Astragali and Radix Angelicae Sinensis induces the expression of erythropoietin in cultured Hep3B cells. Planta Med 2008; 74: 392-395
• 9 Jelkmann W. The role of the liver in the production of thrombopoietin compared with erythropoietin. Eur J Gastroenterol Hepatol 2001; 13: 791-801
• 10 Post DE, Van Meir EG. Generation of bidirectional hypoxia/HIF-responsive expression vectors to target gene expression to hypoxic cells. Gene Ther 2001; 8: 1801-1807
• 11 Ascensao JL, Bilgrami S, Zanjani ED. Erythropoietin: biology and clinical applications. Am J Pediatr Hematol Oncol 1991; 13: 376-387
• 12 Schuster SJ, Badiavas EV, Costagiomi P, Weinmann R, Erslev AJ, Caro J. Stimulation of erythropoietin gene transcription during hypoxia and cobalt exposure. Blood 1989; 73: 13-16
• 13 Goldberg MA, Gaut CC, Schapira L, Antin JH, Ransil BJ, Antman KH, Eder JP, McGarigle CJ, Bunn HF. Erythropoietin gene regulation: From the laboratory to the bedside. Contrib Nephrol 1991; 88: 35-45
• 14 Lönnberg M, Garle M, Lönnberg L, Birgegård G. Patients with anaemia can shift from kidney to liver production of erythropoietin as shown by glycoform analysis. J Pharm Biomed Anal 2013; 81–82: 187-192
• 15 Zheng KYZ, Choi RCY, Xie HQH, Cheung AWH, Guo AJY, Leung KW, Chen VP, Bi CWC, Zhu KY, Chan GKL, Fu Q, Lau DTW, Dong TTX, Tsim KWK. The expression of erythropoietin triggered by Danggui Buxue Tang, a Chinese herbal decoction prepared from Radix Astragali and Radix Angelicae Sinensis, is mediated by the hypoxia-inducible factor in cultured HEK293 T cells. J Ethnopharmacol 2010; 132: 259-267
• 16 Jewell UR, Kvietikova I, Scheid A, Bauer C, Wenger RH, Gassmann M. Induction of HIF-1alpha in response to hypoxia is instantaneous. FASEB J 2011; 15: 1312-1314
• 17 Ratcliffe PJ. HIF-1 and HIF-2: working alone or together in hypoxia?. J Clin Invest 2007; 117: 862-865
• 18 Yeo EJ, Cho YS, Kim MS, Park JW. Contribution of HIF-1alpha or HIF-2alpha to erythropoietin expression: in vivo evidence based on chromatin immunoprecipitation. Ann Hematol 2008; 87: 11-17
• 19 Zheng KYZ, Zhang ZX, Guo AJY, Bi CWC, Zhu KY, Xu SL, Zhan JYX, Lau DTW, Dong TTX, Choi RCY, Tsim KWK. Salidroside stimulates the accumulation of HIF-1α protein resulted in the induction of EPO expression: a signaling via blocking the degradation pathway in kidney and liver cells. Eur J Pharmacol 2012; 679: 34-39
• 20 Ebert BL, Bunn HF. Regulation of the erythropoietin gene. Blood 1999; 94: 1864-1877
• 21 Lee HS, Lin JJ, Kung H, Huang PL, Lee L. The human erythropoietin-encoding gene contains a CAAT box, TATA boxes and other transcriptional regulatory elements in its 5′ flanking region. Gene 1993; 128: 227-236
• 22 La Ferla K, Reimann C, Jelkmann W, Hellwig-Burgel T. Inhibition of erythropoietin gene expression signaling involves the transcription factors GATA-2 and NF-kappa B. FASEB J 2002; 16: 1811-1813
• 23 Roy CN. Anemia of inflammation. Hematology Am Soc Hematol Educ Program 2010; 2010: 276-280
• 24 Yu L, Jiang BP, Luo D, Shen XC, Guo S, Duan JA, Tang YP. Bioactive components in the fruits of Ziziphus jujuba Mill. against the inflammatory irritant action of Euphorbia plants. Phytomedicine 2012; 19: 239-244
• 25 Liu M, Wang Y. cAMP contents of Zizyphus jujuba Mill., Zizyphus spinosus Hu. and other twelve horticultural plants. J Agric Univ Hebei 1991; 14: 20-23
• 26 Batmunkh C, Krajewski J, Jelkmann W, Hellwig-Burgel T. Erythropoietin production: molecular mechanisms of the antagonistic actions of cyclic adenosine monophosphate and interleukin-1. FEBS Lett 2006; 580: 3153-3160
• 27 Boer AK, Drayer AL, Vellenga E. cAMP/PKA-mediated regulation of erythropoiesis. Leuk Lymphoma 2003; 44: 1893-1901
• 28 Winer J, Jung CKS, Shackel I, Williams PM. Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro . Anal Biochem 1999; 270: 41-49