Fo Shou San, an Ancient Herbal Decoction Prepared from Angelicae Sinensis Radix and Chuanxiong Rhizoma, Induces Erythropoietin Expression: A Signaling Mediated by the Reduced Degradation of Hypoxia-Inducible Factor in Cultured Liver Cells

Abstract

Fo Shou San (FSS) is an ancient herbal decoction composed of Angelicae Sinensis Radix (ASR; Danggui) and Chuanxiong Rhizoma (CR; Chuanxiong) in a ratio of 3 : 2. FSS is mainly prescribed for patients having a deficiency of blood supply, and it indeed has been shown to stimulate the production of erythropoietin (EPO) in cultured cells. In order to reveal the mechanism of this FSS-induced EPO gene expression, the upstream regulatory cascade, via hypoxia-induced signaling, was revealed here in cultured hepatocellular carcinoma cell line Hep3B. The induction of EPO gene expression, triggered by FSS, was revealed in cultured hepatocytes by: (i) the increase of EPO mRNA; and (ii) the activation of the hypoxia response element (HRE), an upstream regulator of the EPO gene. The FSS-induced EPO gene expression was triggered by an increased expression of hypoxia-inducible factor-1α (HIF-1α) protein; however, the mRNA expression of HIF-1α was not altered by the treatment of FSS. The increased HIF-1α was a result of reduced protein degradation after the FSS treatment. The current results therefore provide one of the molecular mechanisms of this ancient herbal decoction for its hematopoietic function.

Key words

Angelica sinensis - Ligusticum chuanxiong - Umbelliferae

References

• 1 Bi C W C, Xie H Q H, Xu L, Li J, Cheung A W H, Zhu J T T, Zheng Y Z, Chen V P, Law D T W, Choi R C Y, Wang T J, Dong T T X, Tsim K W K. Fo Shou San, an ancient herbal decoction prepared from Rhizoma Chuanxiong and Radix Angelicae Sinensis, stimulates the production of hemoglobin and erythropoietin in cultured cells.  Planta Med. 2010;  76 1525-1529
• 2 Gregory C J, Eaves A C. Human marrow cells capable of erythropoietic differentiation in vitro: definition of three erythroid colony responses.  Blood. 1977;  49 855-864
• 3 Gregory C J, Eaves A C. Three stages of erythropoietic progenitor cell differentiation distinguished by a number of physical and biologic properties.  Blood. 1978;  51 527-537
• 4 Wu H, Lin X, Jaenisch R, Lodish H F. Generation of committed erythroid BFU-E and CFU-E progenitors does not require erythropoietin or the erythropoietin receptor.  Cell. 1995;  83 59-67
• 5 Lin C S, Lim S K, D'Agati V, Costantini F. Differential effects of an erythropoietin receptor gene disruption on primitive and definitive erythropoiesis.  Genes Dev. 1995;  10 154-164
• 6 Lin F K, Suggs S, Lin C H, Browne J K, Smalling R, Egrie J C, Chen K K, Fox G M, Martin F, Stabinsky Z, Badrawi S M, Lai P H, Goldwasser E. Cloning and expression of the human erythropoietin gene.  Proc Natl Acad Sci USA. 1985;  82 7580-7584
• 7 Lai P H, Everett R, Wang F F, Arakawa T, Goldwasser E. Structural characterization of human erythropoietin.  J Biol Chem. 1986;  261 3116-3121
• 8 Fukuda M N, Sasaki H, Lopez L, Fukuda M. Survival of recombinant erythropoietin in the circulation: the role of carbohydrates.  Blood. 1989;  73 84-89
• 9 Sasaki R, Masuda S, Nagao M. Erythropoietin: multiple physiological functions and regulation of biosynthesis.  Biosci Biotechnol Biochem. 2000;  64 1775-1793
• 10 Fandrey J. Oxygen-dependent and tissue-specific regulation of erythropoietin gene expression.  Am J Physiol Regul Integr Comp Physiol. 2004;  286 R977-R988
• 11 Eckardt K U, Kurtz A. Regulation of erythropoietin production.  Eur J Clin Invest. 2005;  35 13-19
• 12 Warnecke C, Zaborowska Z, Kurreck J, Erdmann V A, Frei U, Wiesener M, Eckardt K U. Differentiating the functional role of hypoxia-inducible factor (HIF)-1α and HIF-2α (EPAS-1) by the use of RNA interference.  FASEB J. 2004;  18 1462-1487
• 13 Goldberg M A, Glass G A, Cunningham J M, Bunn H F. The regulated expression of erythropoietin by two human hepatoma cell lines.  Proc Natl Acad Sci USA. 1987;  84 7972-7976
• 14 Haase V H. Hypoxic regulation of erythropoiesis and iron metabolism.  Am J Physiol Renal Physiol. 2010;  299 F1-F13
• 15 Ebert B L, Bunn H F. Regulation of the erythropoietin gene.  Blood. 1999;  94 1864-1877
• 16 Smith T G, Robbins P A, Ratcliffe P J. The human side of hypoxia-inducible factor.  Br J Haematol. 2008;  141 325-334
• 17 Wang G L, Jiang B H, Rue E A, Semenza G L. Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O₂ tension.  Proc Natl Sci USA. 1995;  92 5510-5514
• 18 Jaakkola P, Mole D R, Tian Y M, Wilson M I, Gielbert J, Gaskell S J, Kriegsheim A V, Hebestreit H F, Mukherji M, Schofield C J, Maxwell P H, Pugh C W, Ratcliffe P J. Targeting of HIF-α to the von Hippel-Lindau ubiquitylation complex by O₂-regulated prolyl hydroxylation.  Science. 2001;  292 468-472
• 19 Ke Q D, Costa M. Hypoxia-inducible factor-1 (HIF-1).  Mol Pharmacol. 2006;  70 1469-1480
• 20 Gao Q T, Cheung J K H, Choi R C Y, Cheung A W H, Li J, Jiang Z Y, Chu G K Y, Duan R, Zhao K J, Dong T T X, Tsim K W K. 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
• 21 Uchimura E, Yamada S, Uebersax L, Fujita S, Miyake M, Miyake J. Method for reverse transfection using gold colloid as a nano-scaffold.  J Biosci Bioeng. 2007;  103 101-103
• 22 Metzen E, Ratcliffe P J. HIF hydroxylation and cellular oxygen sensing.  Biol Chem. 2004;  385 223-230
• 23 Su Y, Loos M, Giese N, Hines O J, Diebold I, Gorlach A, Metzen E, Pastorekova S, Friess H, Buchler P. PHD3 regulates differentiation, tumour growth and angiogenesis in pancreatic cancer.  Br J Cancer. 2010;  103 1571-1579
• 24 Shao M L. Pharmacopoeia of the People's Republic of China. (Part I). Beijing: Chemical Industry Press; 2010: 144
• 25 Liang Q D, Gao Y, Tan H L, Guo P, Li Y F, Zhou Z, Tan W, Ma Z C, Ma B P, Wang S Q. Effects of four Si-Wu-Tang's constituents and their combination on irradiated mice.  Biol Pharm Bull. 2006;  29 1378-1382
• 26 Lin C M, Chiu J H, Wu I H, Wang B W, Pan C M, Chen Y H. Ferulic acid augments angiogenesis via VEGF, PDGF and HIF-1α.  J Nutr Biochem. 2010;  21 627-633
• 27 Ma Z C, Hong Q, Wang Y G, Tan H L, Xiao C R, Liang Q D, Cai S H, Gao Y. Ferulic acid attenuates adhesion molecule expression in gamma-radiated human umbilical vascular endothelial cells.  Biol Pharm Bull. 2010;  33 752-758
• 28 Chan S S K, Cheng T Y, Lin G. Relaxation effects of ligustilide and senkyunolide A, two main constituents of Ligusticum chuanxiong, in rat isolated aorta.  J Ethnopharmacol. 2007;  111 677-680
• 29 Park S S, Bae I, Lee Y J. Flavonoids-induced accumulation of hypoxia-inducible factor (HIF)-1α/2α is mediated through chelation of iron.  J Cell Biochem. 2008;  103 1989-1998
• 30 Zheng K Y Z, Guo A J Y, Bi C W C, Zhu K Y, Chan G K L, Fu Q, Xu S L, Zhan J Y Z, Lau D T W, Dong T T X, Choi R C Y, Tsim K W K. The extract of Rhodiolae Crenulatae Radix et Rhizoma induces the accumulation of HIF-1α via blocking the degradation pathway in cultured kidney fibroblasts.  Planta Med. 2011;  77 894-899
• 31 Zheng K Y Z, Choi R C Y, Xie H Q H, Cheung A W H, Guo A J Y, Leung K W, Chen V P, Bi C W C, Zhu K Y, Chan G K L, Fu Q, Lau D T W, Dong T T X, Zhao K J, Tsim K W K. 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 HEK293T cells.  J Ethnopharmacol. 2010;  132 259-267