甘蔗叶多糖提取工艺的优化及其心肌细胞保护作用的研究  

Optimization of extraction process of polysaccharides from sugarcane leaves and its cardiomyocyte protective effects study

作　　者：王星圆 张帆 黄建朝 邓轩静 郝二伟 杜正彩 李政 廖耀文 张健 侯小涛 邓家刚 WANG Xingyuan;ZHANG Fan;HUANG Jianchao;DENG Xuanjing;HAO Erwei;DU Zhengcai;LI Zheng;LIAO Yaowen;ZHANG Jian;HOU Xiaotao;DENG Jiagang(Faculty of Pharmacy,Guangxi University of Chinese Medicine,Nanning,Guangxi,530200 P.R.China;Guangxi Key Laboratory of TCM Formulas Theory and Transformation for Damp Disease,Guangxi Collaborative Innovation Center of Study on Functional Ingredients of Agricultural Residues,Guangxi Key Laboratory of Eficacy Study on Chinese Materia Medica,Guangxi University of Chinese Medicine,Nanning,Guangxi,530200 P.R.China;Guangxi Fengtang Biochemical Co.,Ltd.,Liuzhou,Guangxi,545000 P.R.China)

机构地区：[1]广西中医药大学药学院,广西南宁530200 [2]广西中医药大学广西中药药效研究重点实验室、广西农作物废弃物功能成分研究协同创新中心、广西中医湿病方药理论与转化重点实验室,广西南宁530200 [3]广西凤糖生化股份有限公司,广西柳州545000

出　　处：《华西药学杂志》2023年第6期621-627,共7页West China Journal of Pharmaceutical Sciences

基　　金：国家自然科学基金资助项目(批准号:82060762);广西科技重大专项(2022AA01016);农作物废弃物功能成分研究协同创新中心(2022-03);广西中药药效研究重点实验室运行补助项目(20-065-38);广西研究生教育创新计划资助项目(YCXJ2021015)。

摘　　要：目的优化甘蔗叶多糖碱性预处理高温提取工艺,分析其单糖组成与分子量,研究其心肌细胞保护作用。方法通过单因素实验和正交试验优化提取工艺。采用1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生化HPLC法分析单糖组成;高效凝胶色谱法检测分子量。采用MTS法检测细胞活力。结果最优提取工艺为:3 mol·L^(-1)碱性溶液预处理30 min,160℃高温提取30 min,提取2次。单糖组成包括甘露糖、鼠李糖、半乳糖醛酸、葡萄糖醛酸、无水葡萄糖、半乳糖、木糖、阿拉伯糖,分子量约30 kD。药效研究发现:甘蔗叶多糖对高糖损伤的心肌细胞具有一定的保护作用。结论碱性预处理高温提取的甘蔗叶多糖含量可达76%;单糖组成分析方法快速稳定、准确简便;甘蔗叶多糖对高糖损伤心肌细胞具有一定的保护作用。OBJECTIVE To optimize the extraction process of polysaccharide from sugarcane leaves,and to analyze its monosaccharide composition and molecular weight,and to study its cardiomyocyte protective effects.METHODS The extraction technology was optimized by Single factor experiment and Orthogonal experiment.The composition of monosaccharides was analyzed by HPLC with PMP pre-column derivatization.Molecular weight was determined by high performance gel chromatography.Cell viability was determined by MTS.RESULTS The optimal extraction process is pretreatment of 3 mol·L^(-1) alkaline solution for 30 min,extraction at 160℃ for 30 min,and extraction twice.Monosaccharides include mannose,rhamnose,galacturonic acid,glucuronic acid,anhydrous glucose,galactose,xylose and arabinose.The molecular weight is approximately 30 kD.The pharmacodynamic study showed that polysaccharides from sugarcane leaf had a protective effect on cardiomyocyte damaged by high glucose.CONCLUSION The polysaccharide content of sugarcane leaves extracted by alkaline pretreatment at high temperature can reach 76%.The method of monosaccharide composition analysis is fast,stable,accurate and simple,and has a certain protective effect on cardiomyocyte damaged by high glucose.

关 键 词：甘蔗叶 多糖 心肌保护 碱性提取 单糖组成 1-苯基-3-甲基-5-吡唑啉酮 H9C2心肌细胞 

分 类 号：R917[医药卫生—药物分析学] R96[医药卫生—药学]