巴戟天多糖的理化性质及其对成骨细胞MC3T3-E1增殖的影响

Physicochemical Properties of Polysaccharides from Morinda officinalis How and Its Influence on the Proliferation of Osteoblasts MC3T3-E1 Cells

以巴戟天为原料,通过酶辅助浸提法提取多糖,采用化学和仪器分析法研究多糖的pH、相对黏度、溶解性、光谱特征、空间构像等理化性质,采用噻唑蓝法测定MC3T3-E1细胞增殖率,酶联免疫法测定Ⅰ型胶原蛋白(collagen typeⅠ,COLⅠ)和骨钙素(osteocalcin,OCN)分泌量及碱性磷酸酶(alkaline phosphatase,ALP)活性,评价多糖提取物对成骨细胞增殖的影响。结果表明,巴戟天多糖属于酸性多糖,pH为6.15±0.13,相对黏度为1.26±0.12,具有糖的特性和三股螺旋结构,溶解性较好,不含蛋白质或多肽;巴戟天多糖提取物浓度为50μg/m L时,MC3T3-E1细胞增殖率极显著增加(P <0.01),达149.67%,COLⅠ和OCN的分泌量极显著增加(P <0.01),分别增加44.98%和85.01%,ALP活性极显著提高(P <0.01),提高了258.95%。表明巴戟天多糖提取物能显著促进MC3T3-E1细胞增殖与分化。

The polysaccharidess was extracted from Morinda officinalis How by enzyme-assisted extraction.The physicochemical properties of polysaccharides such as pH,relative viscosity,solubility,spectral characteristics and spatial conformation were studied by chemical and instrumental analysis.The proliferation rate of MC3T3-E1 cells was determined by Thiazolyl Blue Tetrazolium Bromide( MTT) assay,and the secretion of collagen type( COL Ⅰ),alkaline phosphatase( ALP) and osteocalcin( OCN) were detected by enzyme-linked immunosorbent assay( ELISA).The effect of polysaccharide on the proliferation of osteoblasts was evaluated. The results showed that the polysaccharide of Morinda officinalis How belonged to acidic polysaccharide with pH of 6.15 ± 0.13,relative viscosity of 1.26 ± 0.12,which had sugar characteristics and triple helix structure;had good solubility and did not contain protein or polypeptide.When the extract concentration of Morinda officinalis How polysaccharide was 50 μg/mL,the proliferation rate of MC3T3-E1 cells increased significantly( P < 0.01),to 149.67%,the secretion of COL I and OCN increased significantly( P < 0.01),increased by 44.98% and 85.01% respectively,and the activity of ALP increased significantly( P < 0.01),and increased by 258.95%. These indicated that polysaccharide extracted from Morinda officinalis How could significantly promote the proliferation and differentiation of MC3T3-E1 cells.