臺灣博碩士論文加值系統

血管新生(angiogenesis)在人類的疾病扮演很重要的角色，包括腫瘤生成和心肌梗塞等。VEGF (vascular endothelial growth factor)扮演血管新生重要的分子，血管的生長因子在與血管內皮細胞上的接受器（receptor）結合後，便會活化細胞分泌特定的酵素及生長因子，產生血管修補或新生血管的現象。發炎反應和發炎因子會誘發血管新生現象，目前的研究已經證實發炎物質與新血管生成具有密切的關係，其中對抗新血管作用機轉也漸漸被釐清，包括：interleukin 1 (IL-1)， interleukin 8 (IL-8)， tumor necrosis factor alpha (TNF-α)，interleukin 6 (IL-6)。其中IL-6 與血管新生的關係漸漸被重視，例如心肌梗塞，腫瘤的預後預測。而與VEGF 的相互作用則透過 transcription factor，如STAT3等調控，然而在發炎物質誘導的血管新生作用，可能的基因調控是複雜的而且目前有些部分仍有待釐清。
目前中草藥應用於對抗新血管生成方面，尚待研究。黃芩的主成分黃酮類 (flavonoid)，被認為具有抗發炎的作用，黃芩能否透過抗發炎的作用而達到對抗新血管生成作用的效果，目前仍未知。本論文研究目的是研究黃芩的成分物wogonin和chrysin，透過抗發炎的作用而達到對抗新血管生成作用的效果的作用機轉。利用LPS 或IL-6的誘導，研究wogonin，chrysin透過抑制VEGF/VEGFR 和IL-6/IL-6R自我調控 (autoregulation) 達到對抗血管新生的作用。實驗設計包括內皮細胞運動遷移和血管形成試驗、定量即時聚合酵素連鎖反應(quantitative real-time RT-PCR) 、西方墨點試驗(Western blot analysis)、免疫沉澱西方墨點試驗 (immunoprecipitated Western blot analysis) 和雞胚絨毛尿囊膜 (chicken chorioallantoic membrane，CAM)活體試驗。實驗結果顯示在血管形成型態的改變或是內皮細胞運動遷移的測試，wogonin和chrysin加入後都呈現濃度效應的抑制，在雞胚絨毛尿囊膜活體試驗也呈現相同的抑制效果。而在訊息傳遞的機轉方面，wogonin和chrysin抑制IL-6接受器複合體- gp130，sIL-6Rα，JAK1，STAT3以及VEGF和 VEGF接受器的mRNA及蛋白質的基因表現。藉由STAT3標的siRNA (targeted siRNA) 的感染，STAT3和VEGF之間的傳遞作用，結果顯示wogonin和chrysin都不只能夠抑制單一路徑的訊息傳遞。另一條IL-6的傳遞路徑ERK 1/2蛋白質，細胞內PKC蛋白質表現亦被評估，wogonin和chrysin也呈現抑制作用。
綜合實驗結果，wogonin和chrysin可透過抑制發炎反應達到抑制新血管生成的自我調控;這個結果不僅提供臨床疾病治療的應用和思考，同時也開啟中草藥治療過度新血管生成疾病的契機。

Angiogenesis plays an important role in many human disorders, such as tumor formation and myocardial infarction. In angiogenesis, vascular endothelial growth factor (VEGF) plays a central role and present in response to inflammation, shear stress, and hypoxia. VEGF is involved in the endothelial cell proliferation, motility, and vascular permeability through the binding with the tyrosine kinase receptors (VEGFR-1 and VEGFR-2) expressed in endothelial cells. Several transcription factors, including STAT3, have been shown to up-regulate VEGF gene expression in different cells. Nevertheless, the role of STAT3 in the VEGF regulation on endothelial cells remains unclear. Although, the interplay between IL-6 and VEGF, has been shown in various kinds of cells. The relationship between IL-6 autoregulation and angiogenesis remains to be clarified. In the present investigation, the effect of herbal medicine on anti-angiogenesis remained unclear. Wogonin and chrysin, two pure compounds of Scutellaria baicalensis Georgi, possess potent anti-inflammatory, antimicrobial and anti-oxidant effects. Nonetheless, the role of wogonin or chrysin in angiogenesis, and the potential molecular mechanisms in angiogenesis have not been studied.
The aim of this study was to investigate the effects of wogonin and chrysin on LPS- or IL-6-stimulated angiogenesis. By the treatment with wogonin or chrysin, added into LPS- and IL-6 primarily cultured human umbilical endothelial cells (HUVECs) respectively, the phenotypic alterations were evaluated. Moreover, Western blot analysis, ELISA cytokine assay, immunoprecipitated Western blot analysis and quantitative real time-PCR were performed for IL-6/IL-6R and VEGF/VEGFR gene expressions in vitro. By introducing the targeted siRNA for STAT3, the downstream VEGF signal modulation was also examined. The chicken chorioallantoic membrane (CAM) was applied to test the effects of wogonin and chrysin on LPS- or IL-6-stimulated neovascularization in vivo. The results indicated that wogonin and chrysin (10-8~10-5M) inhibited LPS- or IL-6-stimulated angiogenesis in a concentration-dependent manner in CAM assay. Consequently, negative activation of the mRNA transcription of VEGF/VEGFR-1 and sIL-6Rα/JAK1/STAT3 signaling, as well as the VEGFR-1/VEGFR-2, IL-6, sIL-6Rα, gp130/JAK1 complex protein, were attenuated on wogonin- and chrysin-treated IL-6-stimulated HUVECs, respectively. Besides, another intracellular PKC pathway and alternative intracellular IL-6 receptor signaling pathway, ERK 1/2, were also test and revealed the significant attenuation. Moreover, by introducing targeted siRNA for STAT3, downstream VEGF protein was suppressed on wogonin- and chrysin-treated IL-6-stimulated HUVECs. Taken together, the data conclude that wogonin and chrysin suppress the autoregulation loops of IL-6/IL-6R and VEGF/VEGFR through down-regulation of STAT3, PKC and ERK 1/2 phosphorylation, and hence inhibits LPS- or IL-6-stimulated angiogenesis.
Significantly, wogonin and chrysin were demonstrated to be potential anti-angiogenic agents. The results would bring into perspective the clinical implication for inflammation-related over-angiogenic diseases.

中文摘要 1
Abstract 3
Abbreviations 5
1. Introduction 7
1-1. Angiogenesis and its regulation 7
1-1-1. The formation of angiogenesis 8
1-1-2. Physiological and pathological angiogenesis 9
1-1-3. Factors and cytokines related to angiogenesis 10
1-1-4. Autocrine and paracrine regulation in angiogenesis 11
1-2. Inflammation-stimulated angiogenesis 12
1-2-1. Introduction of acute and chronic inflammation 12
1-2-2. Interaction between inflammation and angiogenesis 14
1-2-3. The role of lipopolysaccharide on inflammation-stimulated angiogenesis 17
1-2-4. The role of interleukin 6 (IL-6) on inflammation-stimulated angiogenesis 18
1-3. Major active ingredients of Scutellaria baicalensis Georgi 18
1-3-1. Effects of Scutellaria baicalensis Georgi on inflammatory diseases 18
1-3-2. Four major active flavonoid ingredients extract from SbG 20
1-4. Rationale and aim of study 20
2. Materials and methods 22
2-1. Instruction equipments 22
2-2. Chemical reagents and cells 23
2-3. Herbal remedy preparation 25
2-3-1. Partial extraction of herbal remedy 25
2-3-2. High performance liquid chromatography (HPLC) 25
2-4. In vitro model 27
2-4-1. Cell cultures 27
2-4-1-1. Preparation of human umbilical veins 27
2-4-1-2. Isolation of human umbilical vein endothelial cells 27
2-4-2. Models for angiogenic phenotypic alteration 28
2-4-2-1. Cell migration assay 28
2-4-2-2. Tube formation assay 29
2-4-3. Cell growth assay for primary HUVEC cultures 30
2-4-3-1. MTT assay for cellular viability 30
2-4-3-2. Trypan blue dye exclusion assay for calculation of cell numbers 30
2-4-4. Models of inflammation-stimulated angiogenesis 31
2-4-4-1. Lipopolysaccharide-stimulated angiogenesis 31
2-4-4-2. Interleukin 6-stimulated angiogenesis 31
2-4-5. Antibody neutralization and chemical inhibition assay for cytokines on HUVECs 31
2-4-6. ELISA assay for cytokine expression levels on HUVECs 32
2-4-7. Signal transduction and protein expressions on HUVECs 32
2-5. In vivo model 36
2-5-1. Experimental models 36
2-5-2. Study protocols 37
2-5-3. Calculation of new vessel formation 38
2-6. Study designs 38
2-7. Statistic 39
3. Results 40
3-1. Effects of Scutellaria baicalensis Georgi and its pure compounds on primary cultured HUVECs by MTT assay 40
3-2. Effects of Scutellaria baicalensis Georgi, wogonin and chrysin on LPS- or IL-6-stimulated HUVECs 40
3-2-1. Effects of Scutellaria baicalensis Georgi, wogonin and chrysin on LPS-stimulated HUVECs 40
3-2-1-1. Effects of Scutellaria baicalensis Georgi, wogonin and chrysin on tube formation and cell migration assays 41
3-2-1-2. Effects of wogonin and chrysin on protein presentation 43
3-2-1-3. Effects of wogonin and chrysin on mRNA transcription 44
3-2-2. Effects of wogonin and chrysin on IL-6-stimulated HUVECs 44
3-2-2-1. Effects of interleukin 6 on cell proliferation and cell viability in HUVEC cultures 44
3-2-2-2. Effects of wogonin and chrysin on tube formation and cell migration assays 45
3-2-2-3. Effects of wogonin and chrysin on protein presentation 46
3-2-2-4. Effects of wogonin and chrysin on mRNA transcription 48
3-3. Effects of wogonin and chrysin on chicken chorioallantoic membrane model in vivo 49
3-3-1. Effects of wogonin and chrysin on chicken chorioallantoic membrane assay in LPS-stimulated model 49
3-3-2. Effects of wogonin and chrysin on chicken chorioallantoic membrane assay in IL-6-stimulated model 49
3-3-3. Calculation of new vessel formation in chicken chorioallantoic membrane model 50
4. Discussion 51
4-1. Experimental models 51
4-1-1. LPS-stimulated angiogenesis 51
4-1-2. IL-6-stimulated angiogenesis as in vitro model 52
4-1-3. Dissimilarities between LPS- and IL-6-stimulated models 52
4-1-4. Chicken chorioallantoic membrane as in vivo model 53
4-2. Discussions of experimental results 54
4-2-1. Cytotoxic effects of wogonin and chrysin on HUVECs 54
4-2-2. Inhibitory effects of wogonin and chrysin on angiogenic phenotypic alteration 55
4-2-3. Modulation of IL-6/IL-6R and VEGF/VEGFR in the effects of wogonin and chrysin on inhibitory angiogenesis 55
4-3. Inhibitory effects of wogonin and chrysin in LPS-stimulated angiogenesis 58
4-3-1. Inhibitory effects of wogonin and chrysin on LPS-stimulated HUVECs 58
4-3-2. Inhibitory modulation of wogonin and chrysin on inflammation and angiogenesis 59
4-4. Inhibitory effects of wogonin and chrysin on IL-6-stimulated angiogenesis 61
4-4-1. Inhibitory effects of wogonin and chrysin on IL-6-stimulated HUVECs 61
4-4-2. Inhibitory modulation of wogonin and chrysin on inflammation and angiogenesis 61
4-4-3. Alternative modulation of wogonin and chrysin on IL-6-stimulated angiogenesis 62
4-5. Similarities and dissimilarities between wogonin and chrysin 63
5. Summary 65
6. Conclusion 67
References 68

1.Aggarwal BB, Bhardwaj A, Aggarwal RS, Seeram NP, Shishodia S, Takada Y. Role of resveratrol in prevention and therapy of cancer: preclinical and clinical studies. Anticancer Res 2004;24:2783-840.
2.Albrecht EA, Chinnaiyan AM, Varambally S, Kumar-Sinha C, Barrette TR, Sarma J. Vidya, et al. PA. Am J Path 2004;106:849-59.
3.Algire GH. An adaptation of the transparent chamber technique to the mouse. J Natl Cancer Inst 1943;4:1-11.
4.Auerbach R, Lewis R, Shinners B, Kubai L, and Akhtar N. Angiogenesis assays: a critical overview. Clin Chem 2003;49:32-40.
5.Augustin HG. Antiangiogenic tumor therapies- will it work? Trends Pharmacol Sci 1998;19:216-22.
6.Bachelot T, Ray-Coquard I, Menetrier-Caux, Rastkha, Duc A, Blay JY. Prognostic value of serum levels of interleukin 6 and serum and plasma levels of vascular endothelial growth factor in hormone-refractory metastatic breast cancer patients. Br J Cancer 2003;88:1721-6.
7.Baluk P, Tammela T, Ator E, Lyubynska N, Achen MG, Hicklin DJ, Jeltsch M et al. Pathogenesis of persistent lymphatic vessel hyperplasia in chronic airway inflammation. J Clin Invest 2005;115:247-57.
8.Beck L Jr, D’Amore PA. Vascular development: cellular and molecular regulation. FASEB J 1997;11:165-73.
9.Bonham M, Posakony J, Coleman I, Montgomery B, Simon J, Nelson PS. Characterization of chemical constituents in Scutellaria baicalensis with antiandrogenic and growth-inhibitory activities toward prostate carcinoma. Clin Cancer Res 2005;11:3905-14.
10.Brooks PC, Montgomery AM, Cheresh DA. Use of the 10-day-old chick embryo model for studying angiogenesis. Methods Mol Biol 1999;129:257-69.
11.Cartwright JE, Johnstone AP, Whitley GS. Endogenously produced nitric oxide inhibits endothelial cell growth as demonstrated using novel antisense cell lines. Br J Pharmacol 2000;131:131-7.
12.Chang CY, Li MC, Liao SL, Huang YL, Shen CC, Pan HC. Prognostic and clinical implication of IL-6 expression in glioma multiforme. J Clin Neurosci 2005;12:930-3.
13.Chang WH, Chen CH, Lu FJ. Different effects of baicalein, baicalin and wogonin on mitochondrial function, glutathione content and cell cycle progression in human hepatoma cell lines. Planta Med 2002;68:128-32.
14.Chang YL, Shen JJ, Wung BS, Cheng JJ, Wang DL. Chinese herbal remedy wogonin inhibits monocyte chemotactic protein-1 gene expression in human endothelial cells. Mol Pharmacol 2001;60:507-13.
15.Chen CC, Chow MP, Huang WC, Lin YC, Chang YJ. Flavonoids inhibit tumor necrosis factor-alpha-stimulated up-regulation of intercellular adhesion molecule-1 (ICAM-1) in respiratory epithelial cells through activator protein-1 and nuclear factor-kappaB: structure-activity relationships. Mol Pharmacol 2004;66:683-93.
16.Chen YC, Shen SC, Chen LG, Lee TJ, Yang LL. Wogonin, baicalin and baicalein inhibition of inducible nitric oxide synthase and cyclooxygenase-2 gene expressions induced by nitric oxide synthase inhibitors and lipopolysaccharide. Biochem Pharmacol 2001;61: 1417-27.
17.Chen YC, Shen SC, Lee WR, Lin HY, Ko CH, Shih CM, et al. Wogonin and fisetin stimulation of apoptosis through activation of caspase 3 cascade and alternative expression of p21 protein in hepatocellular carcinoma cells SK-HEP-1. Arch Toxicol 2002;76:351-9.
18.Chi YS, Lim H, Park H, Kim HP. Effects of wogonin, a plant flavone from Scutellaria radix, on skin inflammation: in vivo regulation of inflammation-associated gene expression. Biochem Pharmacol 2003;66:1271-8.
19.Choi JS, Choi YJ, Park SH, Kong JS, Kang YH. Flavones mitigate tumor necrosis factor-α-induced adhesion molecules upregulation in cultured human endothelial cells: role of nuclear factor-kappa B. J Nut 2004;134:1013-9.
20.Chuang HN, Wang JY, Chiu JH, Tsai TH, Yeh SF, Fu SL, Lui WY et al. Enhancing effects of Scutellaria baicalensis and some of its ingredients on TGF-beta1 geneexpression in RAW264.7 murine macrophage cell line. Planta Med 2005;71:440-5.
21.Colville-Nash PR, Scott DL. Angiogenesis and rheumatoid arthritis: pathogenic and therapeutic implications. Ann Rheum Dis 1992;51:919-25.
22.Coppo P, Flamant S, De MAs V, Jarrier P, Guillier M, Bonnet ML, et al. BCR-ABL activates STAT3 via JAK and MEK pathways in human cells. Br J Haematol 2006;134:171-9.
23.Crane E, List A. Innunomodulatory drugs. Cancer Invest 2005;23:625-34.
24.D’Amore PA, Ng YS. Won’t you be my neighbor? Local stimulation of arteriogenesis. Cell 2002;110:189-92.
25.Dalgleish AG, O’Byrne K. Inflammation and cancer: the role of the immune response and angiogenesis. Cancer treat Res 2006;130:1-38.
26.Dao TT, Kim SB, Sin KS, Kim HP, Park H. Synthesis and biological activities of 8-arylflavones. Arch Pharm Res 2004;27:278-82.
27.Dao TT, Tuyen TN, Kim HP. Synthesis and iNOS inhibitory activities of thioflavones. Arch Pharm Res 2005;28:652-6.
28.de Vries C, Escobedo JA, Ueno H, Houck K, Ferrana N, Williams LT. The fms-like tyrosine kinase, a receptor for vascular endothelial growth factor. Science 1992;255:989-91.
29.Fang J, Xia C, Cao Z, Zheng JZ, Reed E, Jiang BH. Apigenin inhibits VEGF and HIF-1 expression via PI3K/AKT/p70S6K1 and HDM2/p53 pathways. FASEB J 2005;19:342-53.
30.Feghali CA, Wright TM. Cytokines in acute and chronic inflammation. Bioscience 1997;2:12-26.
31.Ferrara N. Role of vascular endothelial growth factor in regulation of physiological angiogenesis. Am J Physiol Cell Physiol 2001;280:C1358-66.
32.Ferrara, N. Vascular endothelial growth factor: Basic science and clinical progress. Endocr Rev 2004;25:581-90.
33.Ferrari D, Pizzirani C, Adinolfi E, Forchap S, Sitta B, Turchet L, Falzoni S et al..The antibiotic polymyxin B modulates P2X7 receptor function. J Immunol 2004;173:4652-60.
34.Folkman J. Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat Med 1995;1:27-31.
35.Fong GH, Rossant J, Gertsenstein M, Breitman ML. Role of the Flt-1 receptor tyrosine kinase in regulation the assembly of vascular endothelium. Nature 1995;376:66-70.
36.Fox SB. Microscopic assessment of angiogenesis in tumors. In: Murray JC, ed. Angiogenesis protocols. Totowa, NJ: Humana Press, 2001;29-46.
37.Fraser HM. Regulation of the ovarian follicular vasculature. Reprod Biol Endocrinol 2006;4:18-26.
38.Gallin JL, Goldstein IM, Snyderman R. Inflammation. Basic principles and clinical correlates, 2ndEd, New York, Raven Press 1992.
39.Gao Z, Huang K, Yang X, Xu H. Free radical scavenging and antioxidant activities of flavonoids extracted from the radix of Scutellaria Baicalensis Georgi. Biochem Biophys Acta 1999;1472: 643-50.
40.Ghosh AK, Hirasawa N, Lee YS, Kim YS, Shin KH, Ryu N, Ohuchi K. Inhibition by acharan suphate of angiogenesis in experimental inflammation models. Br J Pharmacol 2002;137:441-8.
41.Gomez D, Reich NC. Stimulation of primary human endothelial cell proliferation by IFN. J Immunol 2003;170:5373-81.
42.Gong X, Guo C, Huang S, Sun B. Inhaled nitric oxide alleviates hyperoxia suppressed phosphatidylcholine synthesis in endotoxin-induced injury in mature rat lungs. Respir Res 2006;7 in press.
43.Grant, D.S., Tashiro, K., Segui-Real, B., Yamada, Y., Martin, G.R., Kleinman, H.K. Two differentiations of human endothelial cells into capillary-like structures in vitro. Cell 1989;58:933-43.
44.Gupta K, Zhang J. Angiogenesis: a curse or cure? Postgrad Med 2005;81:236-42.
45.Han CK, Son MJ, Chang HW, Chi YS, Park H, Kim HP. Inhibition of prostaglandin production by a structureally-optimized flavonoid derivative, 2, 4,7-trimethoxyflavone and cellular action mechanism. Biol Pharm Bull 2005;28:1366-70.
46.Hatjiharissi E, Terpos E, Papaioannou M, Hatjileontis C, Kaloutsi V, Galaktidou G, et al.. The combination of intermediate doses of thalidomide and dexamethasone reduces bone marrow micro-vessel density but not serum levels of angiogenic cytokines in patients with refractory/relapsed multiple myeloma. Hematol Oncol 2004;22:159-68.
47.Heinrich PC, Behrmann I, Haan S, Hermanns HM, Muller-Newen G, Schaper F. Principles of interleukin (IL)-6- type cytokine signaling and its regulation. Biochem J 2003;374:1-20.
48.Hernandez-Rodriguez J, Sergarra M, Vilardell C, Sanchez M, Garcia-Martinez A, Esteban MJ, Grau JM, Urbano-Marquez A, Colomer D, Kleinman HK, Cid MC. Elevated production of interleukin-6 as associated with a lower incidence of disease-related ischemic events in patients with giant-cell arteritis. Circulation 2003;107:2428-34.
49.Hicklin DJ, Ellis LM. Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis. J Clin Oncol 2005;23:1011-27.
50.Hirota H, Chen J, Betz UA, Rajewsky K, Ross J Jr, Muller W, Chien KR. Loss of a gp130 cardiac muscle cell survival pathway is a crucial event in the onset of heart failure during biomechanical stress. Cell 1999;97:189-98.
51.Hougee S, Sanders A, Faber J, Graus YM, Van Den Berg WB, Graessen J, Smit HF, Hoijer MA. Decreased pro-inflammatory cytokine production by LPS-stimulates PBMC upon in vitro incubation with the flavonoids apigenin, luteolin or chrysin due to selective elimination of monocytes/macrophages. Biochem Pharmacol 2005;15:241-8.
52.Hu CQ, Chen K, Shi Q, Kilkuskie RE, Cheng YC, Lee KH. Anti-AIDS agents, 10. Acacetin-7-O-beta-D-galactopyranoside, an anti-HIV principle from Chrysanthemum morifolium and a structure-activity correlation with some related flavonoids. J Nat Prod 1994;57:42-51.
53.Huang RL, Chen CC, Huang HL, Chang CG, Chen C., Chang C, Hsieh MT. Anti-hepatitis B virus effects of wogonin isolated from Scutellaria baicalensis. Planta Med 2000;66:694-8.
54.Jackson MW, Roberts JS, Heckford SE, Ricciardelli C, Stahl J, Horsfall DJ, Tilley WD. A potential autocrine role for vascular endothelial growth factor in prostate cancer. Cancer Res 2002;62, 854-9.
55.Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 1973;52: 2745-56.
56.Johns A, Zollner S. Angiogenesis in health and disease: basic mechanism and clinical applications. 1st Ed. 2000;P.191-8.
57.Kim HK, Song KS, Park YS, Kang YH, Lee YJ, Lee KR, et al. Elevated levels of circulating platelet microparticles, VEGF, IL-6 and RANTES in patients with gastric cancer: possible role of a metastasis predictor. Eur J Cancer 2003;39:184-91.
58.Kofler S, Nickel T, Weis M. Role of cytokines in cardiovascular diseases: a focus on endothelial responses to inflammation. Clin Science 2005;108:205-13.
59.Kubo M, Kimura Y, Odani T, Tina T, Namba K. Studies on Scutellariae radix. Part II: The antibacterial substance. Planta Med 1981;43:194-201.
60.Kuwano M, Fukushi J, Okamoto M, Nishie A, Goto H, Ishibashi T, Ono M. Angiogenesis factors. Intern Med 2001;40:565-72.
61.Lay IS, Chiu JH, Shiao MS, Lui WY, Wu CW. Crude extract of Salvia mitiorrhiza and Salvianolic acid B enhance in vitro angiogenesis in murine SVR endothelial cell line. Planta Med 2002;68:1-7.
62.Lee H, Kim YO, Kim H, Kim SY, Noh HS, Kang SS, Cho GO et al. Flavonoid wogonin from medicinal herb is neuroprotective by inhibiting inflammatory activation of microglia. FASEB J 2003;17:1943-4.
63.Lee TL, Yeh J, Van Waes C, Chen Z. Epigenetic modification of SOCS-1 differentially regulates STAT3 activation in response to interleukin-6 receptor and epidermal growth factor receptor signaling through JAK and/or MEK in head and neck squamous cell carcinomas. Mol Cancer Ther 2006;5:8-19.
64.Li WW, Talcott KE, Zhai AW, kruger EA, Li VW. The role of therapeutic angiogenesis in tissue repair and regeneration. Adv Skin Wound Care 2005;18:491-500.
65.Lim H, Park H, Kim HP. Inhibition of contact dermatitis in animal models and suppression of proinflammatory gene expression by topically applied flavonoids, wogonin. Arch Pharm Res 2004;27: 442-8.
66.Lin SK, Tsai SC, Lee CC, Wang BW, Liou JY, Shyu KG. Berberine inhibits HIF-1 expression via enhanced proteolysis. Mol Pharmacol 2004;66:612-9.
67.Liu JJ, Huang TS, Cheng WF, Lu FJ. Baicalein and baicalin are potent inhibitors of angiogenesis: inhibition of endothelial cell proliferation, migration and differentiation. Int J Cancer 2003;106:559-65.
68.Loeffler S, Fayard B, Weis J, Weissenberger J. Interleukin-6 induces transcriptional activation of vascular endothelial growth factor (VEGF) in astrocytes in vivo and regulates VEGF promoter activity in glioblastoma cells via direct interaction between STAT3 and Sp1. Int J Cancer 2005;115:202-13.
69.Mahmood N, Pizza C, Aquino R, et al. Inhibition of HIV infection by flavanoids. Antivir Res 1993;22:189-99.
70.Mann DL. Stress activated cytokines and the heart. Cytokines Growth Factor Rev 1996;7:341-54.
71.Maruotti N, Cantatore Fp, Crivellato E, Vacca A, Ribatti D. Angiogenesis in rheumatoid arthritis. Histol Histopathol 2006;21:557-66.
72.Melzig MF, Loose R. Inhibition of lipopolysaccharide (LPS)-induced endothelial cytotoxicity by selected flavonoids. Planta Med 1998;64:397-9.
73.Mezquita J, Mezquita B, Pau M, Mezquita C. Down-regulation of Flt-1 gene expression by the proteasome inhibitor MG262. J Cell Biochem 2003;89:1138-47.
74.Middleton H Jr., Kanadswami C. Effects of flavonoids on immune and inflammatory cell functions. Biochem Pharmacol 1992;43:1167-79.
75.Molostvov G, Morris A, Rose P, Basu S, Muller, G. The effects of selective cytokines inhibiting drugs (CC-10004 and CC-1088) on VEGF and IL-6 expression and apoptosis in myeloma and endothelial cell co-cultures. Br J Haematol 2004;124:366-75.
76.Moon YJ, Wang X, Morris ME. Dietary flavonoids: Effects on xenobiotic and carcinogen metabolism. Toxicol In Vitro 2006; 20:187-210.
77.Murch SJ, Rupasinghe HP, Goodenowe D, Saxena PK. A metabolomic analysis of medicinal diversity in Huang-qin (S Scutellaria baicalensis Georgi) genotypes: discovery of novel compounds. Plant Cell Rep 2004;23:419-25.
78.Nakagawa K, Chen YX, Ishibashi H, Yonemitsu Y, Murata T, Hata Y, Nakashima Y, Sueishi K. Angiogenesis and its regulation: roles of vascular endothelial cell growth factor. Semin Thromb Hemost 2000;26:61-6.
79.Nakamura N, Hayasaka S, Zhang XY, Nagaka Y, Matsumoto M, Hayasaka Y, Terasawa K. Effects of baicalin, baicalein, and wogonin on interleukin-6 and interleukin-8 expression, and nuclear factor-kappa b binding activities induced by interleukin-1 beta in human retinal pigment epithelial cell line. Exp Eye Res 2003;77: 195-202.
80.Naldini A, Carraro F. Role of inflammatory mediators in angiogenesis. Curr Drug Targets Inflamm Allergy 2005;4:3-8.
81.Neilsen PO, Zimmerman GA, McIntyre TM. Escherichia coli Braum lipoprotein induces a lipopolysaccharide-like endotoxic response from primary human endothelial cells. J Immunol 2001;167:5231-9.
82.Nian M, Lee P, Khaper N, Liu P. Inflammatory cytokines and postmyocardial infarction modeling. Circulation Res 2004;94: 1543-53.
83.Nicosia RF, Ottinetti A. Growth of microvessels in serum-free matrix culture of rat aorta. A quantitative assay of angiogenesis in vitro. Lab Invest 1990;63:15-22.
84.Oak MH, El Bedoui J, Schini-Kerth VB. Antiangiogenic properties of natural polyphenols from red wine and green tea. J Nutr Biochem 2005;16:1-8.
85.O' Leary KA, de Pascual-Tereasa S, Needs PW, Bao YP, O'Brien NM, Williamson G. Effect of flavonoids and vitamin E on cyclooxygenase-2 (COX-2) transcription. Mutat Rev 2004;551:245-54.
86.Ono K, Nakane H, Fukushima M, Chermann JC, Barre-Sinoussi F. Differential inhibitory effects of various flavonoids on the activities of reverse transcriptase and cellular DNA and RNA polymerases. Eur J Biochem 1990;190:469-76.
87.Padhani AR, Neeman M. Challenges for imaging angiogenesis. Br J Radio 2001;74:886-90.
88.Paper, D.H. Natural products as angiogenesis inhibitors. Planta Med 1998;64:686-95.
89.Park BK, Heo MY, Park H, Kim HP. Inhibition of TPA-induced cyclooxygenase-2 expression and skin inflammation in mice by wogonin, a plant flavone from Scutellaria radix. Eur J Pharmacol 2001;425:153-7.
90.Park EJ, Zhao YZ, Lian L, Kim YC, Sohn DH. Skullcapflavone I from Scutellaria baicalensis induces apoptosis in activated rat hepatic stellate cells. Planta Med 2005;71:885-7.
91.Park H, Dao TT, Kim HP. Synthesis and inhibition of PGE2 production of 6,8-disubstituted chrysin derivates. Eur J Med Chem 2005;40:943-8.
92.Pepper, M.S., Hazel, S.J., Humel, M., Schleuning, W.D. 8-prenylnaringenin, a novel phytoestrogen, inhibits angiogenesis in vitro and in vivo. J Cell Physiol 2004;199:98-107
93.Piao HZ, Jin SA, Chun HS, Lee JC, Kim WK. Neuroprotective effect of wogonin: potential roles of inflammatory cytokines. Arch Pharm Res 2004;27:930-6.
94.Pollet I, Opina CJ, Zimmerman C, Leong KG, Wong F, Karsan A. Bacterial lipopolysaccharide directly induces Angiogenesis through TRAF6-mediated activation of NF-κB and c-Jun N-terminal kinase. Blood 2003;102:1740-2.
95.Raines EW, Ferri N. Cytokines affecting endothelial and smooth muscle cell in vascular disease. J Lipid Res 2005;46:1081-92.
96.Ramsauer M, D’Amore PA. Getting Tie(2)d up in angiogenesis. J Clin Invest 2002;110:1615-7.
97.Ravindranath MH, Muthugounder S, Presser N, Viswannthan S. Anticancer therapeutic potential of soy isoflavone, genistein. Adv Exp. Med Biol 2004;546:121-65.
98.Resto-Ruiz SI, Schmiederer M, Sweger D, Newton C, Klein TW, Friedman H, Anderson BE. Stimulation of a potential paracrine angiogenic loop between human THP-1 macrophages and human microvascular endothelial cells during Bartonella henselae infection. Infect Immunol 2002;70:4564-70.
99.Risau W. Mechanism of angiogenesis. Nature 1997;386:671-4.
100.Salven P, Hattori K, Heissig B, Rafii S. Interleukin-1α promotes angiogenesis in vivo via VEGFR-2 pathway by inducing inflammatory cell VEGF synthesis and secretion. FESEB J 2002;16:1471-3.
101.Sato Y. Update on endogenous inhibitors of angiogenesis. Endothelium 2006;13:147-55.
102.Schindler R, Mentlein R. Flavonoids and vitamin E reduce the release of the angiogenic peptide vascular endothelial growth factor from human tumor cells. J Nutr 2006;136:1477-82.
103.Shibuya M. Vascular endothelial growth factor receptor family genes: when did the three genes phylogenetically segregates? Biol Chem 2002;383:1573-9.
104.Shieh DE, Liu LT, Lin CC. Antioxidant and free radical scavenging effects of baicalein, bailcalin and wogonin. Anticancer Res 2000;20:2861-5.
105.Sonoda M, Nishiyama T, Matsukawa Y, Moriyasu M. Cytotoxic activities of flavonoids from two Scutellaria plants in Chinese medicine. J Ethnopharmacol 2004;91:65-8.
106.Sueishi K, Yonemitsu Y, Nakagawa K, Kaneda Y, Kumamoto M, Nakashima Y. Atherosclerosis and angiogenesis. Its pathophysiological significance in humans as well as in an animal model induced by the gene transfer of vascular endothelial growth factor. Ann N Y Acad Sci 1997;811:311-22;322-4.
107.Szekanecz Z, Koch AE. Endothelial cells in inflammation and angiogenesis. Curr Drug Targets Inflamm Allergy 2005;4:319-23.
108.Tai MC, Tsang SY, Chang LY, Xue H. Therapeutic potential of wogonin: a naturally occurring flavonoid. CNS Drug Rev 2005;11:141-50.
109.Teo SK. Properties of thalidomide and its analogues: implications for anticancer therapy. AAPS J 2005;7:E14-19.
110.Terman B, Khandke L, Dougher-Vermazan M, Maglione D, Lassam NJ, Gospodarowicz D, et al.. VEGF receptor subtypes KDR and FLT1 show different sensitivities to heparin and placenta growth factor. Growth Factors 1994;11:187-95.
111.Tseng J, Chang JG. Suppression of tumor necrosis factor-alpha, interleukin-1 beta, interleukin-6 and granulocyte-monocyte colony stimulated factor secretion from human monocytes by an extract of Poria cocos. Chung-Hua Min Kuo Wei Sheng Wu Chi Mien I Hsueh Tsa Chih-Chinese J Microbiol Immunol 1992;25:1-11.
112.Unger RE, Peters K, Huang Q, Funk A, Paul D, Kirkpatrick CJ. Vascularization and gene regulation of human endothelial cells growing on porous polyethersulfone (PES) hollow fiber membranes. Biomaterials 2005;26:3461-9.
113.Unger RE, Peters K, Wolf M, Motta A, Migliaresi C, Kirkpatrick CJ. Endothelialization of a non-woven silk fibroin net for use in tissue engineering: growth and gene regulation of human endothelial cells. Biomaterials 2004;25:5137-46.
114.van Dien M, Takahashi K, Mu MM, Koide N, Sugiyama T, Mori I, Yoshida T, Yokochi T. Protective effect of wogonin on endotoxin-induced lethal shock in D-galactosamine-sensitized mice. Microbiol Immunol 2001;45:751-6.
115.van Hinsbergh VW, Engelse MA, Quax PH. Pericellular proteases in angiogenesis and vasculogenesis. Arterioscler Thromb Vasc Biol 2006;26:716-28.
116.von Loon IM. The golden root: Clinical applications of Scutellaria baicalensis Georgi flavonoids as modulators of the inflammatory response. Alt Med Rev 1997;2:472-80.
117.Wang JY, Chuang HN, Chiu JH, Fu SL, Tsai TH, Tsou AP, Ho CP et al.. Effects of Scutellaria baicalensis Georgi on macrophage-hepatocyte interaction through cytokines related to growth control of murine hepatocytes. Exp Biol Med 2006;231:444-55.
118.Weng MS, Ho YS, Lin JK. Chrysin induces G1 phase cell cycle arrest in C6 glioma cells through inducing p21Waf1/Cip1 expression: involvement of p38 mitogen-activated protein kinase. Biochem Pharmacol 2005;69:1815-27.
119.Woo KJ, Jeong YJ, Inoue H, Park JW, Kwon TK. Chrysin suppresses lipopolysaccharide-induced cyclooxygenase-2 expression through the inhibition of nuclear factor for IL-6 (NF-IL6) DNA-binding activity. FEBS Letters 2005;31:705-11.
120.Woo KJ, Jeong YJ, Park Jw, Kwon TK. Chrysin-indcued apoptosis is mediated through caspase activation and Akt inactivation in U937 leukemia cells. Biochem Biophys Res commun 2004;325:1215-22.
121.Woodman OL, Chan ECh. Vascular and anti-oxidant actions of flavonols and flavones. Clin Exp Pharmacol Physiol 2004;31:786-90.
122.Wu H, Zhang G, Minton JE, Ross CR, Blecha F. Regulation of Cathelicidin gene expression: Stimulation by lipopolysaccharide, interleukin-6, retinoic acid, and Salmonella enteica Serovar Typhimurium infection. Infection Immuno 2000;10:5552-8.
123.Xu Q, Briggs J, Park S, Niu G, Kortylewski M, Zhang S, Gritsko T, Turkson J, Kay H, Semenza GL, Cheng JQ, Jove R, Yu H. Targeting Stat3 blocks both HIF-1 and VEGF expression induced by multiple oncogenic signaling pathways. Oncogene 2005;24:5552-60.
124.Yamashiki M, Nishimura A, Suzuki H, Sakaguchi S, Kosaka Y. Effects of the Japanese herbal medicine “Sho-saiko-to” (TJ-9) on in vitro interleukin-10 production by peripheral blood mononuclear cells of patients with chronic hepatitis C. Hepatology 1997;25:1390-7.
125.Yang SH, Lin JK, Huang CJ, Chen WS, Li SY, Chiu JH. Silibinin inhibits angiogenesis via Flt-1, but not KDR, receptor upregulation. J Surg Res 2005;128:140-6.
126.Yeom YJ, Lee HC, Kim GH, Lee HJ, Shim I, Oh SK, Kong SK, Hahm DH. Anti-arthritic effects of Ephedra sinica STAPF herb-acupuncture: Inhibition of lipopolysaccharide-induced inflammation and adjuvant-induced polyarthritis. J Pharmcol Sci 2006;100:41-50.
127.You KM, Jong HG, Kim HP. Inhibition of cyclooxygenase/lipoxygenase from human platelets by polyhydroxylated/methoxylated flavonoids isolated from medical plants. Arch Pharm Res 1999;22:18-24.
128.Young HS, Summers AM, Read IR, Fairhurst DA, Plant DJ, Campalani E, Smith CH et al.. Interaction between Genetic Control of Vascular Endothelial Growth Factor Production and Retinoid Responsiveness in Psoriasis. J Invest Dermatol 2006;126:453-9.
129.Zachary I, Gliki G. Signaling transduction mechanisms mediating biological actions of the vascular endothelial growth factor family. Cardiovas Res 2001;49:568-81.
130.Zhang S, Yang X, Morris ME. Flavonoids are inhibitors of breast cancer resistance protein (ABCG2)-mediated transport. Mol Pharmacol 2004;65:1208-16.