ウシ大網由来血管内皮細胞増殖因子の分離精製と性質の検討
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概要
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The greater omentum has been known to possess a potent angiogenic effect, when it is transposed to the peripheral areas of the body. The factor responsible for the angiogenesis was reported to be present in the lipid fraction extracted from the omentum and was confirmed to be identical to PGE2. However, PGE2 does not enhance the proliferation of vascular endothelial cells in vitro, but is known to generally suppress cell proliferation. In the present study, the endothelial cell growth factor was extracted from the bovine greater omentum, and purification and characterization of the factor were performed. The lipid fraction derived from the bovine omentum exhibited no significant growth-promoting activity when tested with bovine aortic endothelial cells (BAEC). However, the lipids appear to indirectly stimulate neovascularization when injected in vivo. The present work demonstrated the presence of vascular endothelial cell growth factor in the water-soluble extracts of bovine omentum. Gel permeation chromatography of the acid-treated extracts gave two peaks of growth promoting activity apparently corresponding to the molecular weights (MW) of 96,000 and 21,000. The MW 21,000 growth factor is a dominant component, and this factor was partially purified, approximately 120-fold, by ammonium sulfate precipitation (30-80% saturation), acid treatment, hydroxylapatite chromatography, Sephadex G-100 gel exclusion chromatography, and finally phenyl-sepharose chromatography. Heparin affinity chromatography revealed that the factor had no affinity for heparin. The MW 21, 000 growth factor exhibited no additive effect when tested in the presence of heparin. The factor had no growth promoting effect without fetal bovine serum. The activity of MW 21,000 growth factor was stable at trypsin treatment, but inactivated by heating at 100℃ for 2 min, or by acid (pH 2.3) and alkaline (pH 11.2) treatments. These properties clearly distinguish the MW 21,000 growth factor from other well-documented heparin binding growth factor (HBGF). The MW 21,000 growth factor is quite likely distinct from other vascular endothelial cell growth factors, so far described for various tissues and cells. We postulate that the MW 21,000 growth factor detected in the present work contributes to the potent angiogenic activity of the omentum.
- 札幌医科大学の論文
- 1989-02-01
札幌医科大学 | 論文
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