Korean J Physiol Pharmacol.  2009 Apr;13(2):107-113. 10.4196/kjpp.2009.13.2.107.

Olibanum Extract Inhibits Vascular Smooth Muscle Cell Migration and Proliferation in Response to Platelet-Derived Growth Factor

• Choi OB
• Park JH
• Lee YJ
• Lee CK
• Won KJ
• Kim J
• Lee HM
• Kim B

• KMID: 2071659
• DOI: http://doi.org/10.4196/kjpp.2009.13.2.107

Abstract

Olibanum (Boswellia serrata) has been shown to have anti-inflammatory, anti-arthritic and anti- cancer effects. This study determined the role of a water extract of olibanum in platelet-derived growth factor (PDGF)-stimulated proliferation and migration of rat aortic smooth muscle cells (RASMCs). PDGF-BB induced the migration and proliferation of RASMCs that were inhibited by olibanum extract in a dose-dependent manner. The PDGF-BB-increased phosphorylation of p38 mitogen-activated protein kinase (MAPK); the heat shock protein (Hsp) 27 was significantly inhibited by the olibanum extract. The effects of PDGF-BB-induced extracellular signal-regulated kinase1/2 was not altered by the olibanum extract. Treatment with olibanum extract inhibited PDGF-BB-stimulated sprout out growth of aortic rings. These results suggest that the water extract of olibanum inhibits PDGF-BB-stimulated migration and proliferation in RASMCs as well as sprout out growth, which may be mediated by the inhibition of the p38 MAPK and Hsp27 pathways.

Keyword

Olibanum; Migration; Proliferation; Rat aortic smooth muscle cells; Mitogen-activated protein kinase

MeSH Terms

Animals
Boswellia
Cell Movement
Heat-Shock Proteins
Muscle, Smooth, Vascular
Myocytes, Smooth Muscle
p38 Mitogen-Activated Protein Kinases
Phosphorylation
Platelet-Derived Growth Factor
Protein Kinases
Proto-Oncogene Proteins c-sis
Rats
Water
Heat-Shock Proteins
Platelet-Derived Growth Factor
Protein Kinases
Proto-Oncogene Proteins c-sis
Water
p38 Mitogen-Activated Protein Kinases

Figure

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  Fig. 1. Effect of olibanum extract on PDGF-BB-stimulated RASMC migration. The effect of olibanum extract on RASMC migration. The cells from SD rats were treated with or without olibanum extract (1~1,000 μg/ml) or PDGF-BB (10 ng/ml) for 90 min. Migration was quantified using a Boyden chamber assay as described in the Methods section. Cell migration in the quiescent state was expressed as 100% (n=8). Each value represents the mean±SEM. ∗Denotes a significant difference from the PDGF-BB-stimulated state, with p<0.05.

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  Fig. 2. Effect of olibanum extract on PDGF-BB-stimulated RASMC proliferation. Cells were treated with or without olibanum extract (1~1,000 μg/ml) and then stimulated by PDGF-BB (10 ng/ml) for 36 h. Cell proliferation was examined with the BrdU incorporation assay. Proliferation in the quiescent state was considered as 100% (n=10). Each value represents the mean±SEM. ∗Significant differences from responses in the PDGF-BB-stimulated state, with p<0.05.

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  Fig. 3. Effects of olibanum extract on activity of MAPK in RASMCs. After cells were treated without or with olibanum extract (1 ~ 1,000 μg/ml) for 30 min, cells were stimulated with PDGF-BB (10 ng/ml) for 10 min. The RASMC lysates were immunoblotted with antibodies. The phosphorylation of p38 MAPK and ERK1/2 was examined using phosphor-specific antibodies. The total expression of kinases and β-actin was measured using non-phospho-specific antibodies and anti-β-actin antibody, respectively. (B, C) Statistical analysis of the phosphorylation level of MAPKs obtained in (A). The basal levels of phosphorylation are considered as 100% (n=4). Each value represents the mean±SEM. ∗Significant differences from the PDGF-BB-stimulated state, with p<0.05. p-p38, phosphorylated p38 MAPK; p-ERK1/2, phosphorylated ERK1/2.

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  Fig. 4. Effect of olibanum extract on PDGF-BB-induced phosphorylation of Hsp27 in RASMCs. The phosphorylation of Hsp27 was examined using a phospho-specific antibody. (B) Statistical analysis of the phosphorylation level of Hsp27 obtained in (A). The basal levels of phosphorylation are considered as 100% (n=4). Each value represents the mean±SEM. ∗Significant differences from the PDGF-BB-stimulated state, with p<0.05. p-Hsp27, phosphorylated Hsp27.

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  Fig. 5. Ex vivo analysis of olibanum extract on sprout formation of aortic rings in response to PDGF-BB. Aortic rings (1 mm) were embedded in Matrigel and cultured. Rings without any treatment (vehicle) and PDGF-BB (10 ng/ml) or olibanum extract (1~1,000 μg/ml)-treated rings. Results were obtained on day 5. (B) The statistical results obtained from panel (A). The level in vehicle (0.1% DMSO)- treated rings is expressed as 100% (n=4). Each value represents the mean±SEM. ∗Denotes a significant difference from the PDGF-BB (10 ng/ml)-stimulated state, with p<0.05.

Reference

Bilato C., Pauly RR., Melillo G., Monticone R., Gorelick-Feldman D., Gluzband YA., Sollott SJ., Ziman B., Lakatta EG., Crow MT. Intracellular signaling pathways required for rat vascular smooth muscle cell migration. Interactions between basic fibroblast growth factor and platelet-derived growth factor. J Clin Invest. 96:1905–1915. 1995.

Champagne MJ., Dumas P., Orlov SN., Bennett MR., Hamet P., Tremblay J. Protection against necrosis but not apoptosis by heat-stress proteins in vascular smooth muscle cells: evidence for distinct modes of cell death. Hypertension. 33:906–913. 1999.

Chevrier MR., Ryan AE., Lee DY., Zhongze M., Wu-Yan Z., Via CS. Boswellia carterii extract inhibits TH1 cytokines and promotes TH2 cytokines in vitro. Clin Diagn Lab Immunol. 12:575–580. 2005.

Dardik A., Yamashita A., Aziz F., Asada H., Sumpio BE. Shear stress-stimulated endothelial cells induce smooth muscle cell chemotaxis via platelet-derived growth factor-BB and interleukin-1alpha. J Vasc Surg. 41:321–331. 2005.

Gayathri B., Manjula N., Vinaykumar KS., Lakshmi BS., Balakrishnan A. Pure compound from Boswellia serrata extract exhibits anti-inflammatory property in human PBMCs and mouse macrophages through inhibition of TNFalpha, IL-1beta, NO and MAP kinases. Int Immunopharmacol. 7:473–482. 2007.

Geraldes P., Sirois MG., Bernatchez PN., Tanguay JF. Estrogen regulation of endothelial and smooth muscle cell migration and proliferation: role of p38 and p42/44 mitogen-activated protein kinase. Arterioscler Thromb Vasc Biol. 22:1585–1590. 2002.

Grotendorst GR., Seppä HE., Kleinman HK., Martin GR. Attachment of smooth muscle cells to collagen and their migration toward platelet-derived growth factor. Proc Natl Acad Sci USA. 78:3669–3672. 1981.

Guay J., Lambert H., Gingras-Breton G., Lavoie JN., Huot J., Landry J. Regulation of actin filament dynamics by p38 map kinase-mediated phosphorylation of heat shock protein 27. J Cell Sci. 110:357–368. 1997.

Gupta I., Gupta V., Parihar A., Gupta S., Lüdtke R., Safayhi H., Ammon HP. Effects of Boswellia serrata gum resin in patients with bronchial asthma: results of a double-blind, placebo-controlled, 6-week clinical study. Eur J Med Res. 3:511–514. 1998.

Hedges JC., Dechert MA., Yamboliev IA., Martin JL., Hickey E., Weber LA., Gerthoffer WT. A role for p38 MAPK/HSP27 pathway in smooth muscle cell migration. J Biol Chem. 274:24211–24219. 1999.

Huang MT., Badmaev V., Ding Y., Liu Y., Xie JG., Ho CT. Anti-tumor and anti-carcinogenic activities of triterpenoid, beta-boswellic acid. Biofactors. 13:225–230. 2000.

Kim B., Kim J., Bae YM., Cho SI., Kwon SC., Jung JY., Park JC., Ahn HY. p38 mitogen-activated protein kinase contributes to the diminished aortic contraction by endothelin-1 in DOCA-salt hypertensive rats. Hypertension. 43:1086–1091. 2004.

Kimmatkar N., Thawani V., Hingorani L., Khiyani R. Efficacy and tolerability of Boswellia serrata extract in treatment of osteoarthritis of knee-a randomized double blind placebo controlled trial. Phytomedicine. 10:3–7. 2003.

Kozawa O., Tanabe K., Ito H., Matsuno H., Niwa M., Kato K., Uematsu T. Sphingosine 1-phosphate regulates heat shock protein 27 induction by a p38 MAP kinase-dependent mechanism in aortic smooth muscle cells. Exp Cell Res. 250:376–380. 1999.

Lee CK., Lee HM., Kim HJ., Park HJ., Won KJ., Roh HY., Choi WS., Jeon BH., Park TK., Kim B. Syk contributes to PDGF-BB-mediated migration of rat aortic smooth muscle cells via MAPK pathways. Cardiovas Res. 74:159–168. 2007.

Lee HM., Jeon BH., Won KJ., Lee CK., Park TK., Choi WS., Bae YM., Kim HS., Lee SK., Park SH., Irani K., Kim B. Gene transfer of redox factor-1 inhibits neointimal formation: involvement of platelet-derived growth factor-beta receptor signaling via the inhibition of the reactive oxygen species-mediated Syk pathway. Circ Res. 104:219–227. 2009.

Lee HM., Kim HJ., Park HJ., Won KJ., Kim J., Shin HS., Park PJ., Kim HJ., Lee KY., Park SH., Lee CK., Kim B. Spleen tyrosine kinase participates in Src-mediated migration and proliferation by PDGF-BB in rat aortic smooth muscle cells. Arch Pharm Res. 30:761–769. 2007.

Liu B., Ryer EJ., Kundi R., Kamiya K., Itoh H., Faries PL., Sakakibara K., Kent KC. Protein kinase C-δ regulates migration and proliferation of vascular smooth muscle cells through the extracellular signal-regulated kinase 1/2. J Vascs Surg. 45:160–168. 2007.

Millette E., Rauch BH., Kenagy RD., Daum G., Clowes AW. Platelet-derived growth factor-BB transactivates the fibroblast growth factor receptor to induce proliferation in human smooth muscle cells. Trends Cardiovasc Med. 16:25–28. 2006.

Nicocia RF., Ottinetti A. Growth of microvessels in serum-free matrix culture of rat aorta. Lab Invest. 63:115–122. 1990.

Pearson G., Robinson F., Beers Gibson T., Xu BE., Karandikar M., Berman K., Cobb MH. Mitogen-activated protein (MAP) kinase pathways: regulation and physiological functions. Endocr Rev. 22:153–183. 2001.

Pompili VJ., Gordon D., San H., Yang Z., Muller DW., Nabel GJ., Nabel EG. Expression and function of a recombinant PDGF B gene in porcine arteries. Arterioscler Thromb Vasc Biol. 15:2254–2264. 1995.

Raines EW., Ross R. Smooth muscle cells and the pathogenesis of the lesions of atherosclerosis. Br Heart J. 69:S30–37. 1993.

Robinson MJ., Cobb MH. Mitogen-activated protein kinase pathways. Curr Opin Cell Biol. 9:180–186. 1997.

Ross R. Atherosclerosis is an inflammatory disease. New Engl J Med. 340:115–126. 1999.

Salinthone S., Ba M., Hanson LJ., Martin JL., Halayko AJ., Gerthoffer WT. Overexpression of human Hsp27 inhibits serum induced proliferation in airway smooth muscle myocytes and confers resistance to hydrogen peroxide cytotoxicity. Am J Physiol Lung Cell Mol Physiol. 293:L1194–L1207. 2007.

Sharma ML., Khajuria A., Kau A., Singh S., Singh GB., Atal CK. Effect of salai guggal ex-Boswellia serrata on cellular and humoral immune responses and leucocyte migration. Agents Actions. 24:161–164. 1988.

Wei CY., Chou YH., Ho FM., Hsieh SL., Lin WW. Signaling pathways of LIGHT induced macrophage migration and vascular smooth muscle cell proliferation. J Cell Physiol. 209:735–743. 2006.

Winking M., Sarikaya S., Rahmanian A., Jödicke A., Böker DK. Boswellic acids inhibit glioma growth: a new treatment option? J Neurooncol. 46:97–103. 2000.

Y J., Kamath JV., Asad M. Effect of hexane extract of Boswellia serrata oleo-gum resin on chemically induced liver damage. Pak J Pharm Sci. 19:129–133. 2006.

Zhan Y., Kim S., Izumi Y., Izumiya Y., Nakao T., Miyazaki H., Iwao H. Role of JNK, p38, and ERK in platelet-derived growth factor-induced vascular proliferation, migration, and gene expression. Arterioscler Thromb Vasc Biol. 23:795–801. 2003.