酶解等4种方法用于中药废弃物资源化研究初探(Ⅰ)——以脉络宁注射液生产中石斛药渣的多糖资源化研究为例  

Recycling of Wastes from Traditional Chinese Medicine Production by Wet Ultrafine Grinding Technology and Other Three Methods(Ⅰ)——Recycling of Polysaccharide in Mailuoning Injection Production Waste Dendrobii Mobile Dregs

作　　者：刘双双[1,2] 刘丽芳[1] 朱华旭[2] 郭立玮[2] 段金廒[2] 潘永兰[2] 徐向前 

机构地区：[1]中国药科大学天然药物活性组分与药效国家重点实验室,南京210009 [2]南京中医药大学江苏省中药资源产业化过程协同创新中心,南京210023 [3]金陵药业股份有限公司南京金陵制药厂研究所,南京210009

出　　处：《中国实验方剂学杂志》2016年第22期34-40,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江苏省科技厅自然科学基金项目(BK2012855);江苏省科技厅产学研联合创新资金--前瞻性研究项目(BY2012036);江苏省六大人才高峰项目(2014-YY-014);江苏省中药资源产业化过程协同创新中心分离平台建设项目(ZDXM-1-6;ZDXM-1-9);江苏省高校优势学科建设工程资助项目;南京中医药大学校级创新团队项目

摘　　要：目的：通过4种不同提取方法用于中药生产中废弃物的多糖资源化的比较研究,优选出脉络宁注射液生产废弃物石斛药渣中提取多糖的方法,实现对中药资源的再循环利用。方法：采用酶解法、湿法超微粉碎法、超声法和加热回流法分别提取石斛药渣中石斛多糖,以苯酚-硫酸法测定其多糖提取率;通过单因素考察和正交试验对提取率最高的提取方法进行工艺优选;考察石斛药渣多糖（DDP）对小鼠脾淋巴细胞的代谢转化和小鼠单核巨噬细胞RAW264.7释放一氧化氮（NO）能力的影响。结果：4种提取方法的多糖提取率分别为0.51%,8.79%,5.34%,3.44%,确定最佳提取方法为酶解;其最佳酶解条件：p H5.3,料液比1∶55,温度50℃,酶活性4 000 U·g-1,提取时间300 min;与对照组比较,质量浓度在62.5~125 mg·L-1,DDP显著促进小鼠脾淋巴细胞增殖,DDP与脂多糖（LPS）协同促进作用显著（P〈0.05,P〈0.01）,质量浓度为62.5 mg·L-1时,DDP与Con A协同促进作用显著（P〈0.05）,质量浓度在62.5~500 mg·L-1,DDP对小鼠单核巨噬细胞RAW264.7产生NO的促进作用极显著（P〈0.01）。结论：酶解法提取石斛药渣中多糖的工艺简便、得率稳定,具免疫增强作用,具有推广应用价值。Objective： To compare four different extraction methods for recycling wastes from traditional Chinese medicine production,in order to select the optimal method to extract polysaccharide from Mailuoning injection production waste Dendrobii Mobile dregs. Method： The four methods,namely wet ultrafine grinding method,enzyme method,ultrasonic method,heating reflux method,were adopted. The polysaccharide extraction rate was detected by phenol sulfuric acid colorimetry; the process with the highest extraction rate was determined by single factor and orthogonal test; the effect of Dendrobii Mobile dregs polysaccharide（DDP） on the proliferation and transformation of lymphocytes in mouse and the NO release of macrophage RAW264. 7 were investigated.Result： The polysaccharide extraction rates of Dendrobii Mobile dregs by the above four different methods were0. 51%,8. 79%,5. 34%,3. 44%, respectively. The optimal extraction method was determined to be the enzymolysis; its optimal enzymolysis conditions were p H at 5. 3,solid-liquid ratio at 55,temperature at 50 ℃,enzyme dosage of 4 000 U·g- 1,extraction time of 300 minutes. Compared with control group,DDP significantly enhanced the proliferation of splenic lymphocytes at concentrations between 62. 5-125 mg·L- 1,with an obvious synergistic effect between DDP and lipopolysaccharide（LPS）（P〈 0. 05,P〈 0. 01）; at the concentration of 62. 5mg·L- 1,DDP showed a significant synergistic effect with Con A（P 〈0. 05）; at concentrations between 62. 5-500mg·L- 1,DDP played a notable role in promoting NO release of mouse macrophage RAW264. 7（P 〈0. 01）.Conclusion： Enzyme extraction process of polysaccharide from Dendrobii Mobile dregs was simple and stable,with the effect in immune enhancement,and worth clinical promotion.

关 键 词：脉络宁注射液 石斛药渣 多糖 资源化 酶解法 免疫活性 

分 类 号：R282[医药卫生—中药学]