蚁狮多糖的提取工艺及体内免疫活性研究

Study on the Extraction Technology of Polysaccharides from Antlion and Its Immune Activity in vivo

目的:优化蚁狮多糖的提取工艺,探究其对小鼠免疫功能的影响。方法:以蚁狮多糖含量为考察指标,比较水提取法、蛋白酶水解提取法(以酶用量、提取温度、提取时间为考察因素进行正交试验优选)和稀碱液提取法(以碱液浓度、提取温度、提取时间为考察因素进行正交试验优选)提取蚁狮多糖的效果。取128只KM小鼠随机分为4个研究组,每个研究组随机分为对照组(等体积生理盐水)与蚁狮多糖低、中、高剂量组(20、40、80 mg/kg),每组8只,尾静脉iv给药,0.2 mL/10 g,每日1次,连用1周,分别用于测定小鼠腹腔巨噬细胞吞噬百分率与吞噬指数、脾和胸腺指数、淋巴细胞转化率和血清溶血素水平。结果:3种方法所得提取液中蚁狮多糖含量分别为14.48%、38.66%、30.62%。蛋白酶水解提取法所得蚁狮多糖含量最高,其最优提取工艺条件为40℃下用100μg/g的木瓜蛋白酶提取3 h。与对照组比较,蚁狮多糖低、中、高剂量组小鼠腹腔巨噬细胞吞噬百分率和吞噬指数、脾指数显著升高(P<0.05),胸腺指数显著降低(P<0.05),而淋巴细胞转化率无显著变化(P>0.05);蚁狮多糖中剂量组小鼠血清溶血素水平显著升高(P<0.05)。结论:蛋白酶水解提取法适于蚁狮多糖的提取,其优选工艺可靠。蚁狮多糖具有增强小鼠非特异性免疫、体液免疫的活性。

OBJECTIVE：To optimize the extraction technology of polysaccharides from antlion and explore its effect on immune functions of mice. METHODS：Using content of polysaccharides as investigation index,the effects of extracting polysaccharides from antlion by water extraction method protease hydrolysis extraction（optimized by orthogonal test using extraction temperature,enzyme dosage,extraction time as indexes）,and diluted alkali extraction（optimized by orthogonal test using alkali concentration,extraction temperature,extraction time as indexes）were compared. 128 KM mice were randomly divided into 4 groups,then randomly divided into control group（normal saline）,polysaccharides low-dose,medium-dose,high-dose groups（20,40,80 mg/kg）,8 in each group,iv in tail vein,0.2 mL/10 g,once a day,for 1 week,which were respectively used to determine the phagocytosis percentage and phagocytic index of peritoneal macrophages,spleen and thymus index,lymphocyte transformation rate and serum hemolysin levels. RESULTS：The contents of polysaccharides by 3 methods were 14.48%,38.66%,30.62%,respectively.The content of polysaccharides by protease hydrolysis extraction was the highest,the optimal extraction technology were as follows as using 100 μ g/g papain extracting 3 h under 40 ℃. Compared with control group,phagocytosis percentage,phagocytic index,spleen index in polysaccharides low-dose,medium-dose,high-dose groups were significantly increased（P〈0.05）,thymus index was significantly decreased（P〈0.05）,while lymphocyte transformation rate had no significant changes（P〉0.05）;serum hemolysin in polysaccharides medium-dose group was significantly increased（P〈0.05）. CONCLUSIONS：Protease hydrolysis extraction is suitable for the extraction of polysaccharides from antlion,the optimal technology is reliable. Polysaccharides from antlion show activity in enhancing mice non-specific immunity and humoral immunity.