中药组合物多糖的提取工艺及抗氧化活性研究  

Extraction Technology and Anti-oxidant Activity of Polysaccharide from Traditional Chinese Medicine Compositions

作　　者：谭超杰 王博 邹文奇 任晶 刘静颐 李春阳 白丽丹 盛瑜[1] TAN Chaojie;WANG Bo;ZOU Wenqi;REN Jing;LIU Jingyi;LI Chunyang;BAI Lidan;SHENG Yu(School of Pharmacy,Beihua University,Jilin 132013,China;Zunyi Medical University Zhuhai Campus,Zhuhai 519041,China)

机构地区：[1]北华大学药学院,吉林吉林132013 [2]遵义医科大学珠海校区,广东珠海519041

出　　处：《北华大学学报（自然科学版）》2022年第2期218-224,共7页Journal of Beihua University（Natural Science）

基　　金：国家自然科学基金项目(82004067);吉林省科技厅省级医药健康产业发展专项资金项目(20170311039YY);北华大学国家级大学生创新创业计划项目(201910201044).

摘　　要：目的优化中药(北虫草-刺五加)抗疲劳组合物中多糖的提取工艺,并探讨其体外抗氧化活性.方法通过单因素试验、正交试验法优化北虫草-刺五加多糖(Cordyceps militaris-Acanthopanax senticosus polysaccharide,CAP)的提取工艺;通过体外1,1-二苯基-2-苦肼基(1,1-diphenyl-2-picrylhydrazyl,DPPH)清除率、总超氧阴离子自由基(total superoxide anion free radical,SAFR)清除率和羟基自由基(hydroxyl free radical)清除率比较CAP与北虫草多糖(Cordyceps militaris polysaccharides,CMP)、刺五加多糖(Acanthopanax senticosus polysaccharide,ASPS)的抗氧化能力差异;利用H_(2)O_(2)诱导大鼠肾上腺髓质嗜铬瘤分化细胞(PC12细胞),构建体外氧化损伤模型,从而考察三者体外抗氧化作用差异.结果提取温度90℃、提取时间3 h、料液比m(样品,g)∶V(水,mL)=1∶40、提取次数3次为CAP的最佳提取工艺,多糖得率可达27.61%;CAP对DPPH的清除率最高可达到63.02%,对SARF的清除率最高达到67.52%,对HFR的清除率最高达到68.38%,均高于CMP和ASPS;100μg/mL的CAP可显著提高H_(2)O_(2)损伤的PC12细胞的存活率,并且可以显著提高PC12细胞中超氧化物歧化酶(Superoxide dismutase,SOD)和过氧化氢酶(Catalase,CAT)活力,显著降低PC12细胞中丙二醛(Malondialdehyde,MDA)含量,作用均高于同等浓度的CMP和ASPS.结论该提取方法稳定可靠,获得的组合物多糖CAP具有明显的体外抗氧化活性,且能够通过减弱H_(2)O_(2)产生的活性氧而保护PC12细胞免受损伤,作用效果优于组合物中的各味中药多糖.Objective To optimize the extraction process of polysaccharide from anti-fatigue compositions of traditional Chinese medicine(Cordyceps militaris-Acanthopanax senticosus),and to expolre its anti-oxidant activity in vitro.Method The extraction process of Cordyceps militaris-Acanthopanax senticosus polysaccharide(CAP)was optimized by single factor experiment and orthogonal experiment method;The difference of antioxidant capacity between CAP,Cordyceps polysaccharide militaris polysaccharides(CMP)and Acanthopanax senticosus polys-accharide(ASPS)was compared by in vitro 1,1-diphenyl-2-picrazine(1,1)-diphenyl-2-picrylhydrazyl(DPPH)scavenging rate,total superoxide anion free radical(SAFR)scavenging rate and hydroxyl free radical scavenging rate;H_(2)O_(2) was used to induce rat adrenal medullary pheochromoma cells(PC12 cells)to construct oxidative damage model in vitro to investigate the difference in anti-oxidant effects in vitro.Results The extraction temperature was 90℃,the extraction time was 3 h,the material-liquid ratio was 1∶40 g/mL,and the extraction times were 3 times.The optimal extraction process for CAP was that the yield of polysaccharides could reach 27.61%;The highest clearance rate of CAP on DPPH can reach 63.02%,the highest clearance rate for SARF is 67.52%,and the highest clearance rate for hydroxyl free radical is 68.38%,both are higher than CMP and ASPS;100μg/mL CAP can significantly improve the survival rate of PC12 cells damaged by H_(2)O_(2),and can significantly increase the activity of superoxide dismutase(SOD)and catalase(CAT)in PC12 cells,and signi-ficantly reduce the content of malondialdehyde(MDA)in PC12 cells.The effects are higher than the same conc-entration of CMP and ASPS.Conclusion The extraction method is stable and reliable,the obtained composition polysaccharide CAP has obvious antioxidant activity in vitro,and can protect PC12 cells from damage by reducing the reactive oxygen species(ROS)produced by H_(2)O_(2),and the effect is better than all the single traditional medicine p

关 键 词：北虫草-刺五加组合物 抗疲劳 多糖 提取工艺 抗氧化 

分 类 号：R284[医药卫生—中药学]