1.Tropical Medicine Institute, Guangzhou University of Chinese Medicine, Guangzhou 510405, China;
2. The 2nd Guangdong Hospital of Traditional Chinese Medicine, Guangzhou 510095, China
Abstract��
OBJECTIVE To disclose the pathological mechanism of bacterial LPS-induced angiogenesis and hyperplasia in tumor-like hyperplasia and the pharmacological mechanism underlying artesunate and betulilic acid blocking tumor-like hyperplasia. METHODS The tumor-like hyperplasia models were established for mouse hypodermis and articular synovium using LPS or LPS-containing complete Fround′s adjuvant (CFA). The morphological features (inflammatory grades) were described and the histochemical signatures (angiogenesis, hyperplasia, and inflammatory infiltration) were examined. The serial concentrations of nitric oxide (NO), blood oxygen saturation (SpO2) and 3-nitrotyrosine (3NT) were quantitatively determined by biochemical, physiological and immunological procedures, and the expression levels of inducible nitric oxide synthase (iNOS), hypoxia-induced factor 1α (HIF-1α) and vascular endothelial growth factor (VEGF) were analyzed by the immunohistochemical method. RESULTS Both LPS and CFA can lead to the inflammatory phenotypes and abnormal hyperplasia-related microscopic alterations. They enable the elevation of NO, decline of SpO2, and increase of 3NT (protein nitration). The considerable up-regulation of iNOS, HIF-1α and VEGF are accompanied by angiogenesis and hyperplasia. While a NO donor compound replicates such pathogenesis, a NO synthesis inhibitor antagonizes this effect. Artesunate and betulilic acid down-regulate iNOS, HIF-1α and VEGF, decrease NO, and increase SpO2, thereby leading to the melioration or interruption of aberrant inflammatory angiogenesis and hyperplasia. CONCLUSION LPS-triggered iNOS overexpression and potent NO burst may represent the direct drivers for angiogenesis and hyperplasia. Due to repression of the pathogenic process of tumor-like hyperplasia, artesunate and betulilic acid possess the prophylactic and therapeutic potentials.
GAO Qian-,
WU Pei-,
HE Jiang- etc
.Artesunate and Betulilic Acid Block Lipopolysaccharide-Induced Angiogenesis and Hyperplasia in Mice[J] Chinese Pharmaceutical Journal, 2015,V50(4): 330-338
��
[1]
WU Y S, CHEN S N. Inflammation and carcinogenesis . J Cancer Ther, 2013, 4(10): 1449-1451.
[2]
THE EUROGAST STUDY GROUP. An international association of Helicobacter pylori infection and gastric cancer . Lancet, 1993, 341(8857): 1359-1362.
[3]
IARC WORKING GROUP ON THE EVALUATION OF CARCINOGENIC RISKS TO HUMANS. Schistosomes, liver flukes and Helicobacter pylori . IARC Monogr Eval Carcinog Risks Hum, 1994, 61(1): 177-240.
[4]
SCHMAUSSER B, ANDRULIS M, ENDRICH S, et al. Expression and subcellular distribution of toll-like receptors TLR4, TLR5 and TLR9 on the gastric epithelium in Helicobacter pylori infection . Clin Exp Immunol, 2004, 136(3):521-526.
[5]
KENTARO C, TAKASHI I, MANABU K, et al. Helicobacter pylori augments growth of gastric cancers via the lipopolysaccharide-Toll-like receptor 4 pathway whereas its lipopolysaccharide attenuates antitumor activities of human mononuclear cells . Clin Cancer Res, 2008, 14(10): 2909-2917.
[6]
TONINO P. Gastritis and Gastric Cancer - New Insights in Gastroprotection, Diagnosis and Treatments . Rijeka:InTech, 2011.
[7]
BAO F, WU P, XIAO N, et al. Nitric oxide-driven hypoxia initiates synovial angiogenesis, hyperplasia and inflammatory lesions in mice . PLoS One, 2012, 7(3): 34494.
[8]
HULTQVIST M, OLOFSSON P, GELDERMAN K A, et al. A new arthritis therapy with oxidative burst inducers. PLoS Med, 2010, 3(9): 348.
[9]
WANG J X, TANG W, ZHOU R, et al. The new water-soluble artemisinin derivative SM905 ameliorates collagen-induced arthritis by suppression of inflammatory and Th17 responses . Br J Pharmacol, 2008, 153(6): 1303-1310.
[10]
TEACHEY D T, GREINER R, SEIF A, et al. Treatment with sirolimus results in complete responses in patients with autoimmune lymphoproliferative syndrome . Br J Haematol, 2009, 145(1): 101-106.
[11]
WU P, BAO F, XIAO N, et al. Artemisinin and rapamycin compromise nitric oxide-driven and hypoxia-triggered acute articular synovitis in mice . Sci Sin Vit (�й���ѧ:������ѧ), 2012, 42(9): 724-738.
[12]
CUI J, MAO X, OLMAN V, et al. Hypoxia and miscoupling between reduced energy efficiency and signaling to cell proliferation drive cancer to grow increasingly faster . J Mol Cell Biol, 2012, 4(3): 174-176
[13]
YOSHIMOTO T, NAKANISHI K, HIROSE S, et al. High serum IL-6 level reflects susceptible status of the host to endotoxin and IL-1/tumor necrosis factor . J Immunol, 1992, 148(11): 3596-3603.
[14]
ZENG L P, WEN J K, WEI S Z. Effects of liposaccharide on nitric oxide synthase gene expression in different tissues of rat . Chin J Arteriosclerosis (�й�����Ӳ����־), 1997, 5(2): 112-115.
[15]
LIU C F, JIANG J M, LIANG P Z, et al. Inhibitive effect of aminoguanidine on the expression of nitric oxide in osteoarthritic synoviocytes induced by lipopolysaccharide . J Clin Rehab Tiss Eng Res (�й���֯�����о����ٴ�����), 2007, 11(4): 689-691.
[16]
NODA T, AMANO F. Differences in nitric oxide synthase activity in a macrophage-like cell line, RAW264.7 cells, treated with lipopolysaccharide (LPS) in the presence or absence of interferon-gamma (IFN-gamma): Possible heterogeneity of iNOS activity . J Biochem, 1997, 121(1): 38-46.
[17]
HARRISON D G, BATES J. The nitrovasodilators:New ideas about old drugs . Circulation, 1993, 87(5): 1461-1467.
[18]
WENG X H, CHEN S. Infection and multiple organ failure: Infectious diseases . New Med (��ҽѧ), 1996, 27(11): 605-607.
[19]
XU W, CHARLES I G, MONCADE S. Nitric oxide: Orchestrating hypoxia regulation through mitochondrial respiration and the endoplasmic reticulum stress response . Cell Res, 2005, 15(1): 63-65.
[20]
NG C T, BINIECKA M, KENNEDY A, et al. Synovial tissue hypoxia and inflammation in vivo . Ann Rheum Dis, 2010, 69(7): 1389-1395.
[21]
LU M P, DU L Z, YU Z Z, et al. Meconium induces formation of nitrotyrosine and expression of inducible nitric oxide synthase in the rat lung . Chin J Pathophysiol (�й�����������־), 2005, 21(8): 1472-1475.
[22]
NATARAJAN R, FISHER B J, FOWLER A A 3RD. Regulation of hypoxia inducible factor-1 by nitric oxide in contrast to hypoxia in microvascular endothelium . FEBS Lett, 2003, 549(1-3): 99-104.
[23]
SABER T, VEALE D J, BALOGH E, et al. Toll-like receptor 2 induced angiogenesis and invasion is mediated through the Tie2 signalling pathway in rheumatoid arthritis . PLoS One, 2011, 6(8): 23540.
[24]
OHNISHI K, SEMI K, YAMAMOTO T, et al. Premature termination of reprogramming in vivo leads to cancer development through altered epigenetic regulation . Cell, 2014, 156(4): 663-677.
[25]
WANG W Q, ZHOU H J, CHEN H H, et al. Inhibitory effect of artesunate on angiogenesis . Chin J Pharmacol Toxicol (�й�ҩ��ѧ�붾��ѧ��־), 2004, 18(1): 32-36.
[26]
SUN J H, WANG H. Effects of artesunate on rat corneal neovascularization . Chin J Ocul Trauma Occupat Eye Dis (������ְҵ�۲���־), 2007, 29(3): 172-175.
[27]
ZHOU H J, WANG W Q, WU G D, et al. Artesunate inhibits angiogenesis and downregulates vascular endothelial growth factor expression in chronic myeloid leukemia K562 cells . Vascul Pharmacol, 2007, 47(2-3): 131-138.
[28]
LIANG A H, XUE B Y, LI C Y, et al. Inhibitory effects of artesunate on endotoxin-induced nitric oxide biogenesis . China J Chin Mater Med (�й���ҩ��־), 2001, 26(11): 770-773.
[29]
HUANG L, ZHANG P Z, ZENG Q P. Studies on artemisinin sensitizing compound in inhibiting tumor cell proliferation and suppressing transplanted tumor growth in tumor-bearing nude mice . J Guangzhou Univ Chin Med (������ҩ��ѧѧ��), 2010, 27(3): 254-257.
[30]
EIZNHAMER D A, XU Z Q. Betulinic acid: A promising anticancer candidate . Idrugs, 2004, 7(4): 359-373.
[31]
GAO Y, JIA Z, KONG X, et al. Combining betulinic acid and mithramycin a effectively suppresses pancreatic cancer by inhibiting proliferation, invasion, and angiogenesis . Cancer Res, 2011, 71(15): 5182-5193.
[32]
KARNA E, SZOKA L, PALKA J A. Betulinic acid inhibits the expression of hypoxia-inducible factor-1a and vascular endothelial growth factor in human endometrial adenocarcinoma cells . Mol Cell Biochem, 2010, 340(1-2): 15-20.
[33]
YI J E, OBMINSKA-MRUKOWICZ B, DU J Y, et al. Effects of betulinic acid on immune function and antioxidant activity of mouse peritoneal macrophages . Acta Nutr Sin (Ӫ��ѧ��), 2010, 32(3): 281-285.
[34]
CHINTHARLAPALLI S, PAPINENI S, RAMAIAH S K, et al. Betulinic acid inhibits prostate cancer growth through inhibition of specificity protein transcription factors . Cancer Res, 2007, 67(6): 2816-2823.
[35]
STEINKAMP-FENSKE K, BOLLINGER L, XU H, et al. Reciprocal regulation of endothelial nitric-oxide synthase and NADPH oxidase by betulinic acid in human endothelial cells. J Pharmacol Exp Ther, 2007, 322(2): 836-842.
2015, Vol. 50 
