Yagi A, Egusa T, Arase M, Tanabe M, Tsuji H
Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, Hiroshima, Japan.
Planta Med. 1997 Feb;63(1):18-21. doi: 10.1055/s-2006-957595.
Fractions of leaf gel from Aloe barbadensis Mill. were prepared by gel permeation using DEAE Sephadex A-25, Sepharose 6B, and Sephadex G-50 columns. These were then tested by in vitro assays for proliferation of human normal dermal and baby hamster kidney cells. The glycoprotein fraction promoted cell growth, while the neutral polysaccharide fraction did not show any growth stimulation. Moreover, the polar-colored glycoprotein fraction strongly inhibited the in vitro assays. An active glycoprotein fraction (protein 82%, carbohydrate 11%) examined on polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE showed a single band. Its molecular weight was 29 kD on a Sephadex G-50 column and its isoelectric point was pH 6.8. Immunoblotting after SDS-PAGE showed that the glycoprotein was composed of two subunits (14 kD). Deglycosylation of glycoprotein (Pg21-2b fraction) by trifluoromethanesulphonic acid provided a protein band with a molecular weight of 13 kD on SDS-PAGE. The colored glycoprotein fraction was shown on SDS-PAGE to be a mixture with a molecular weight of 18 kD-15 kD. It was later hydrolyzed with 10% H2SO4 to produce phenolic substances.
采用DEAE葡聚糖A - 25、琼脂糖6B和葡聚糖G - 50柱，通过凝胶渗透法制备了库拉索芦荟叶凝胶的组分。然后通过体外试验检测这些组分对人正常皮肤细胞和幼仓鼠肾细胞增殖的影响。糖蛋白组分促进细胞生长，而中性多糖组分未显示出任何生长刺激作用。此外，极性有色糖蛋白组分强烈抑制体外试验。在聚丙烯酰胺凝胶电泳（PAGE）和SDS - PAGE上检测的活性糖蛋白组分（蛋白质82%，碳水化合物11%）显示为单一条带。在葡聚糖G - 50柱上其分子量为29 kD，等电点为pH 6.8。SDS - PAGE后的免疫印迹显示该糖蛋白由两个亚基（14 kD）组成。用三氟甲磺酸对糖蛋白（Pg21 - 2b组分）进行去糖基化处理后，在SDS - PAGE上得到一条分子量为13 kD的蛋白条带。有色糖蛋白组分在SDS - PAGE上显示为分子量为18 kD - 15 kD的混合物。后来用10% H₂SO₄水解产生酚类物质。
