北苍术粗多糖提取工艺优化及体外抗炎活性研究

Optimization of extraction process and in vitro anti-inflammatory activities of crude polysaccharides from A.chinensis (DC.) Koidz

目的利用北苍术为实验原料,优化其粗多糖的提取工艺,并对其体外抗炎活性进行研究。方法选用干燥北仓术块茎,用无水乙醇进行脱脂处理,制备脱脂粗多糖粉。苯酚硫酸法检测多糖含量,以多糖得率为指标,利用正交实验确定酶法辅助超声提取北仓术粗多糖(ACCPS)的最佳工艺。采用Elson-Morgan改良法检测透明质酸酶的活性,观测ACCPS对透明质酸酶的抑制效果。在酸性条件下,利用考马斯亮蓝法检测加入不同浓度ACCPS溶液后牛血清蛋白质含量的变化。结果得到葡萄糖标准曲线为y=0.963 7x+0.028 0。最优提取工艺为:酶比m纤维素酶∶m果胶酶=1∶2,酶用量为1.2%,酶解时间为30 min,酶解pH为6.5,酶解温度为40℃,超声时间为40 min,超声功率为80%,此时粗多糖得率17.72%。在研究体外抗炎活性的实验中,透明质酸酶的抑制率随ACCPS溶液质量浓度的增加而增加,在2.500 mg/mL时抑制率达到76.21%。得到蛋白质标准曲线为y=0.023 0x+0.021 5。随着ACCPS溶液质量浓度的增加,对牛血清白蛋白的抑制作用逐渐增加。在ACCPS溶液质量浓度为4 mg/mL时,最高的抑制率为63.75%。结论 ACCPS具有较好的抗炎活性。

Objective To optimize extraction process of Atractylodes chinensis crude polysaccharides(ACCPS),and study in vitro anti-inflammatory activity of A.chinensis(DC.)Koidz.Methods Dried glutinous tubers were selected and degreased with absolute ethanol to prepare defatted crude polysaccharide powder.The polysaccharide content was determined by phenol sulfuric acid method,and optimal yield of polysaccharides from ACCPS by enzymatic assisted ultrasound method was determined through orthogonal design.The activity of hyaluronidase was measured by Elson-Morgan modified method,and inhibitory effect of ACCPS on hyaluronidase was observed.Under acidic conditions,the Coomassie Brilliant Blue method was used to detect changes of bovine serum protein content after addition of different concentrations of ACCPS solution.Results The glucose standard curve was obtained as y=0.963 7 x+0.028 0.The optimal extraction process were as follows:enzyme ratio mcellulase∶mpectinase=1∶2,enzyme dosage was 1.2%,enzymatic hydrolysis time was 30-minute,enzymatic hydrolysis pH was 6.5,enzymatic hydrolysis temperature was 40℃,ultrasonic time was 40-minute,ultrasonic power was 80%,and crude polysaccharide yield was 17.72%.The inhibition rate of hyaluronidase increased with the increase of ACCPS solution concentration,and inhibition rate reached 76.21%at 2.500 mg/mL.The protein standard curve was obtained as y=0.023 0 x+0.021 5.With concentration of ACCPS solution increased,the inhibition effect on bovine serum albumin was gradually increased.When concentration of ACCPS solution was 4 mg/mL,the highest inhibition rate was 63.75%.Conclusion It is demonstrated that ACCPS showed good anti-inflammatory activity.