鲫鱼卵唾液酸糖蛋白制备工艺优化及其促骨形成作用研究

以14种水产动物卵为原料,筛选出具有较高促前成骨MC3T3-E1细胞增殖活性的鲫鱼卵水溶性糖蛋白,采用响应面法优化了鲫鱼卵唾液酸糖蛋白(Sialoglycoprotein of Carassius auratus eggs,Ca-SGP)的提取工艺,并评价Ca-SGP对MC3T3-E1细胞增殖、分化和矿化的影响。结果表明,14种水产动物卵中,鲫鱼卵糖蛋白唾液酸含量最高,且对MC3T3-E1细胞增殖率最高,分别为5.76%和145.71%;采用单因素和响应面试验优化得到制备Ca-SGP的最优提取工艺为:NaCl浓度为0.38 mol/L、提取时间为4.2 h、液料比为1:3.88 w/v,在此条件下,Neu5Ac得率的预测值为592 mg/100 g,验证值为588.47 mg/100 g。Ca-SGP对骨形成作用的研究结果表明,Ca-SGP显著提高MC3T3-E1细胞增殖至135.56%,显著提高COL I、OCN和OPN的分泌量,分别提高到32.23、261.46和85.89 ng/mL,显著提高成骨细胞矿化能力至241.67%,说明Ca-SGP具有显著促骨形成作用(P<0.05)。以上研究为寻找安全、有效抗骨质疏松功能因子提供新途径。

Study on Optimization of Sialyglycoproteins Extraction from Carassius auratus Egg and Its Osteogenic Activity

Water-soluble glycoprotein of Carassius auratus eggs,which exhibited well MC3T3-E1 cell proliferation activity,was screened from 14 kinds of aquatic animal eggs. Response surface method was employed to optimize the extraction condition of sialoglycoprotein from Carassius auratus eggs(Ca-SGP),and its effects on MC3T3-E1 cell proliferation,differentiation and mineralization were evaluated. The results showed that the Neu5Ac content and MC3T3-E1 cell proliferation rate of water-soluble glycoprotein from Carassius auratus eggs were the highest among 14 kinds of aquatic animal eggs,which were 5.76% and 145.71%,respectively. Single factor and response surface experiment were used to optimize the optimal extraction process of Ca-SGP,and the optimal extraction process were NaCl concentration 0.38 mol/L,extraction time 4.2 h,and the liquid-material ratio 1:3.88 w/v. Under these conditions,the predicted value of Neu5Ac yield was 592 mg/100 g,and the verified value was 588.47 mg/100 g.The results of osteogenic activity of Ca-SGP showed that Ca-SGP significantly increased the proliferation of MC3T3-E1 cell to 135.56%,the secretion of COL I,OCN and OPN to 32.23,261.46 and 85.89 ng/mL,respectively,and the mineralization ability of MC3T3-E1 to 241.67%,which indicating that Ca-SGP could significantly promote bone formation(P<0.05).These above studies provide a new way for finding safe and effective functional factors of anti-osteoporosis.