Journal List > Korean J Physiol Pharmacol > v.15(1) > 1025773

Hwang and Kim: Inhibitory Effects of Corni Fructus Extract on Angiogenesis and Adipogenesis

Abstract

Natural products in Chonnam, Korea were screened via anti-angiogenesis experiments, and 1 candidate product was identified, Corni fructus, which exerted dose-dependent inhibitory effects against angiogenesis, adipogenesis, and cell adhesion. C. fructus extract (CFE) exhibits an angiogenesis inhibitory effect superior to that of the EGCG from green tea leaves. The expression level of angiogenesis and adipogenesis-related signal molecules in the western blotting was reduced by increasing the amount of added CFE. Moreover, a diet supplemented with CFE was deemed more effective in inducing weight loss in LB mice than a representative synthetic diet drug, orlistat, which incidently caused the side effect of denuding the mice of their hair. These results indicate that C. fructus may prove to be a useful anti-adipogenic compound, and these in vitro results may be reflected later under in vivo conditions.

References

1. Han DH, Kim SK, Kang SW, Choe BK, Kim KS, Chung JH. Matrix metallopeptidase 2 gene polymorphism is associated with obesity in Korean population. Korean J Physiol Pharmacol. 2008; 12:125–129.
crossref
2. Jung MY, Kim BS, Kim YJ, Koh IS, Chung JH. Assessment of relationship between fyn-related kinase gene polymorphisms and overweight/obesity in Korean population. Korean J Physiol Pharmacol. 2008; 12:83–87.
crossref
3. Pataky Z, Bobbioni-Harsch E, Golay A. Obesity: a complex growing challenge. Exp Clin Endocrinol Diabetes. 2010; 118:427–433.
crossref
4. Frezza EE, Wachtel MS, Ewing BT. The impact of morbid obesity on the state economy: an initial evaluation. Surg Obes Relat Dis. 2006; 2:504–508.
crossref
5. Folkman J, Shing Y. Angiogenesis. J Biol Chem. 1992; 267:10931–10934.
crossref
6. Doldi N, Bassan M, Gulisano M, Broccoli V, Boncinelli E, Ferrari A. Vascular endothelial growth factor messenger ribonucleic acid expression in human ovarian and endometrial cancer. Gynecol Endocrinol. 1996; 10:375–382.
crossref
7. Cao Y. Angiogenesis modulates adipogenesis and obesity. J Clin Invest. 2007; 117:2362–2368.
crossref
8. Crandall DL, Hausman GJ, Kral JG. A review of the micro-circulation of adipose tissue: anatomic, metabolic, and angiogenic perspectives. Microcirculation. 1997; 4:211–232.
crossref
9. Hausman GJ, Richardson RL. Adipose tissue angiogenesis. J Anim Sci. 2004; 82:925–934.
10. Neels JG, Thinnes T, Loskutoff DJ. Angiogenesis in an in vivo model of adipose tissue development. FASEB J. 2004; 18:983–985.
crossref
11. Voros G, Maquoi E, Demeulemeester D, Clerx N, Collen D, Lijnen HR. Modulation of angiogenesis during adipose tissue development in murine models of obesity. Endocrinology. 2005; 146:4545–4554.
crossref
12. Christiaens V, Lijnen HR. Angiogenesis and development of adipose tissue. Mol Cell Endocrinol. 2010; 318:2–9.
crossref
13. Rudkowska I, Roynette CE, Demonty I, Vanstone CA, Jew S, Jones PJ. Diacylglycerol: efficacy and mechanism of action of an anti-obesity agent. Obes Res. 2005; 13:1864–1876.
crossref
14. Obeid OA, Bittar ST, Hwalla N, Emery PW. Effect of diet supplementation with glutamine, dihydroxyacetone, and leucine on food intake, weight gain, and postprandial glycogen metabolism of rats. Nutrition. 2005; 21:224–229.
crossref
15. Prestwich TC, Macdougald OA. Wnt/beta-catenin signaling in adipogenesis and metabolism. Curr Opin Cell Biol. 2007; 19:612–617.
16. Yun JW. Possible anti-obesity therapeutics from nature–a review. Phytochemistry. 2010; 71:1625–1641.
17. Oh S, Kim KS, Chung YS, Shong M, Park SB. Anti-obesity agents: a focused review on the structural classification of therapeutic entities. Curr Top Med Chem. 2009; 9:466–481.
crossref
18. Lee SO, Kim SY, Han SM, Kim HM, Ham SS, Kang IJ. Corni fructus scavenges hydroxy radicals and decreases oxidative stress in endothelial cells. J Med Food. 2006; 9:594–598.
crossref
19. Choi WH, Chu JP, Jiang MH, Baek SH, Park HD. Effects of fraction obtained from Korean Corni Fructus extracts causing anti-proliferation and p53-dependent apoptosis in A549 lung cancer cells. Nutr Cancer. 2011; 63:121–129.
crossref
20. Park CH, Noh JS, Tanaka T, Yokozawa T. Effects of morroniside isolated from Corni Fructus on renal lipids and inflammation in type 2 diabetic mice. J Pharm Pharmacol. 2010; 62:374–380.
crossref
21. Yamabe N, Noh JS, Park CH, Kang KS, Shibahara N, Tanaka T, Yokozawa T. Evaluation of loganin, iridoid glycoside from Corni Fructus, on hepatic and renal glucolipotoxicity and inflammation in type 2 diabetic db/db mice. Eur J Pharmacol. 2010; 648:179–187.
crossref
22. Ramírez-Zacarías JL, Castro-Muñozledo F, Kuri-Harcuch W. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with Oil red O. Histochemistry. 1992; 97:493–497.
23. Lee DH, MacIntyre JP, Wang E, Hudson DJ, Ishaque A, Conant JA, Pope BL, Lau CY. A leukocyte lipid up-regulates the avidity of lymphocyte function-associated antigen-1. Biochem Biophys Res Commun. 1994; 199:319–326.
crossref
24. Vandermeeren M, Janssens S, Borgers M, Geysen J. Dimethylfumarate is an inhibitor of cytokine-induced E-selectin, VCAM-1, and ICAM-1 expression in human endothelial cells. Biochem Biophys Res Commun. 1997; 234:19–23.
crossref
25. Carmeliet P, Lampugnani MG, Moons L, Breviario F, Compernolle V, Bono F, Balconi G, Spagnuolo R, Oosthuyse B, Dewerchin M, Zanetti A, Angellilo A, Mattot V, Nuyens D, Lutgens E, Clotman F, de Ruiter MC, Gittenberger-de Groot A, Poelmann R, Lupu F, Herbert JM, Collen D, Dejana E. Targeted deficiency or cytosolic truncation of the VE-cadherin gene in mice impairs VEGF-mediated endothelial survival and angiogenesis. Cell. 1999; 98:147–157.
crossref
26. Zhang D, Bar-Eli M, Meloche S, Brodt P. Dual regulation of MMP-2 expression by the type 1 insulin-like growth factor receptor: the phosphatidylinositol 3-kinase/Akt and Raf/ERK pathways transmit opposing signals. J Biol Chem. 2004; 279:19683–19690.
27. Ballinger A, Peikin SR. Orlistat: its current status as an anti-obesity drug. Eur J Pharmacol. 2002; 440:109–117.
crossref
28. Jandacek RJ, Woods SC. Pharmaceutical approaches to the treatment of obesity. Drug Discov Today. 2004; 9:874–880.
crossref
29. Birari RB, Bhutani KK. Pancreatic lipase inhibitors from natural sources: unexplored potential. Drug Discov Today. 2007; 12:879–889.
crossref
30. Weigle DS. Pharmacological therapy of obesity: past, present, and future. J Clin Endocrinol Metab. 2003; 88:2462–2469.
crossref
31. Choi YS, Park H, Jeong S. Distinct role of PI3-kinase/Akt pathway in the activation of etoposide-induced NF- B transcription factor. J Microbiol Biotechnol. 2006; 16:391–398.
32. Tang FY, Nguyen N, Meydani M. Green tea catechins inhibit VEGF-induced angiogenesis in vitro through suppression of VE-cadherin phosphorylation and inactivation of Akt molecule. Int J Cancer. 2003; 106:871–878.
33. Tang FY, Meydani M. Green tea catechins and vitamin E inhibit angiogenesis of human microvascular endothelial cells through suppression of IL-8 production. Nutr Cancer. 2001; 41:119–125.
crossref

Fig. 1.
Anti-angiogenesis experiment of 18 natural products with the positive control, EGCG. HUVEC was cultured on Matrigel-coated wells, and the tube formation of HUVEC cells was randomly photographed using a digital camera, and analyzed by the Scion Image software.
kjpp-15-43f1.tif
Fig. 2.
Cytotoxicity in HUVEC cells with different concentrations of C. fructus extract (CFE). HUVEC was seeded onto 96-well plates and treated with various doses (0.1, 0.5, 1, 5, 10, 25, 50, and 100 mg/l) of CFE for 48 hrs. MTT solution was added to the wells, and the 96-well plates were incubated. The absorbances of the 96-well plates were measured with a microplate reader at 540 nm. The value was converted to cell viability based on the control.
kjpp-15-43f2.tif
Fig. 3.
CFE was subjected to preparative size exclusion column of Asahipak GS-310. CFE was chromatographed on an Asahipak GS-310 column eluted with methanol at a flow rate of 5 ml/min, and monitored at 307 nm. CFE was separated into six fractions. Each fraction was evaluated via anti-angiogenesis experiments. HUVEC was cultured on Matrigel-coated wells, and the tube formation of HUVEC cells was randomly photographed using a digital camera, and analyzed by the Scion Image software.
kjpp-15-43f3.tif
Fig. 4.
Anti-angiogenesis effect of SHE fractions 1, 5, and 6. Anti-angiogenesis experiments were conducted with different concentrations (1, 5, 10, and 25 ppm) of fractions 1, 5, and 6. The pictures show the cellular morphology of HUVEC cells treated with different concentrations of fractions 1, 5, and 6.
kjpp-15-43f4.tif
Fig. 5.
Various doses (1, 5, 10, and 20 mg/l) of CFE addition on U937 cell adhesion. (A) Cell adhesion of U937 on IL-1β and CFE stimulated HUVEC, (B) Cell adhesion of PMA- and CFE-stimulated U937 on HUVEC, (C) Cell adhesion of CFE-stimulated U937 on IL-1β- and CFE-stimulated HUVEC cells. Each treatment in the cell adhesion experiments was conducted as described in the METHODS section.
kjpp-15-43f5.tif
Fig. 6.
CFE inhibits the interaction of PI3-kinase with VE-cadherin and Akt upon cell activation with VEGF. Proteins from HUVEC cells were immunoprecipitated with PI3-kinase antibody and immunoblotted with antibodies to VE-cadherin and Akt. β-actin was employed as a positive control.
kjpp-15-43f6.tif
Fig. 7.
The effect of CFE addition in the course of 3T3-L1 adipocyte differentiation. 3T3-L1 was treated with CFE at various concentrations (1, 5, 10, and 25 mg/l). After differentiation, lipid accumulation was stained with Oil red O solution and the morphological changes were observed via microscopy. The Oil red O stained lipid was quantified with a microplate reader at 520 nm.
kjpp-15-43f7.tif
Fig. 8.
(A) CFE inhibits the interaction of PI3-kinase with VE-cadherin and Akt upon cell activation with VEGF. Proteins from HUVEC cells were immunoprecipitated with PI3-kinase antibody and immunoblotted with antibodies to VE-cadherin and Akt. β-actin was employed as a positive control. Expression of PPARγ on 3T3-L1 preadipocyte with CFE (A) and expression of SREBP-1 and PPARγ on 3T3-L1 adipocytes with CFE (B). Protein from 3T3-L1 was treated with different concentrations (1, 5, and 10 mg/l) of CFE, and extracted with RIPA buffer for western blotting.
kjpp-15-43f8.tif
Fig. 9.
Weight change of LB mice fed for 8 weeks on a CFE-supplemented diet. The inset shows the side effects of the synthetic obesity agent, orlistat.
kjpp-15-43f9.tif
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