百合地黄汤多糖提取工艺优化及抗焦虑抑郁药效评价  

Optimization of extraction process of polysaccharide from Baihe dihuang decoction and study on its antianxiety and anti-depression efficacy

作　　者：唐林[1] 赵洪庆[2] 杨蕙[3] 刘检[3] 何怡然 李莉[1] 张水寒[5] TANG Lin;ZHAO Hongqing;YANG Hui;LIU Jian;HE Yiran;LI Li;ZHANG Shuihan(Hospital-made Preparations Center,the First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Science&Technology Innovation Center,Hunan University of Chinese Medicine,Changsha 410208,China;Medical Innovation Experiment Center,the First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Dept.of Pharmacy,the First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Hunan Academy of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学第一附属医院制剂中心,长沙410007 [2]湖南中医药大学科技创新中心,长沙410208 [3]湖南中医药大学第一附属医院医学创新实验中心,长沙410007 [4]湖南中医药大学第一附属医院药学部,长沙410007 [5]湖南省中医药研究院,长沙410208

出　　处：《中国药房》2023年第8期923-928,共6页China Pharmacy

基　　金：湖南省自然科学基金资助项目(No.2022JJ80093);湖南省教育厅科学研究项目(No.22B0362);湖南中医药大学中医学一流学科开放基金项目(No.2021ZYX27);湖南中医药大学校级科研基金项目(No.2019XJJJ055);湖南省高新技术产业科技创新引领计划项目(No.2020SK2029)。

摘　　要：目的 优化百合地黄汤多糖提取工艺并初步评价其抗焦虑抑郁药效。方法 在Plackett-Burman实验基础上,以多糖得率与多糖含量的综合评分为评价指标,提取时间、加水量、醇沉浓度为考察因素,结合Box-Behnken响应面法优化百合地黄汤多糖提取工艺并验证。将40只ICR小鼠分为对照组、文法拉辛组[阳性对照,13.5 mg/(kg·d)]和百合地黄汤多糖高、低剂量组[5.28、2.64g/(kg·d),以生药量计],每组10只(雌雄各半)。药物组小鼠灌胃相应药液,对照组小鼠灌胃水10 mL/kg,每天1次,连续7 d。采用高架十字迷宫实验、旷场实验、强迫游泳实验、悬尾实验评价最优工艺所制提取物对小鼠焦虑抑郁样行为的改善情况,并采用酶联免疫吸附测定法检测其对小鼠脑组织中神经递质水平的影响。结果 百合地黄汤多糖的最优提取工艺为加25倍量水,提取1.5 h,醇沉浓度70%。3次验证实验的多糖得率、多糖含量平均值分别为33.10%、0.62 mg/mg,与预测值(36.14%、0.65 mg/mg)的相对偏差分别为8.40%、4.62%(RSD<2%,n=3)。所得百合地黄汤多糖提取物能显著增加小鼠进入开放臂的次数比例及开放臂停留时间比例、自主活动总距离及中心区域活动距离,并显著缩短其强迫游泳实验、悬尾实验不动时间(P<0.05或P<0.01);可显著升高小鼠脑组织中5-羟色胺、去甲肾上腺素(百合地黄汤多糖低剂量组除外)、γ氨基丁酸水平,并显著降低谷氨酸(百合地黄汤多糖低剂量组除外)水平(P<0.05或P<0.01)。结论 优化的百合地黄汤多糖提取工艺稳定、可行;所得多糖提取物具有明显的体内抗焦虑抑郁作用。OBJECTIVE To optimize the extraction process and to primarily evaluate the anti-anxiety and anti-depression efficacy of polysaccharide from Baihe dihuang decoction.METHODS Based on Plackett-Burman experimental design,using the comprehensive score of yield and content of polysaccharide as indicators,with extraction time,water amount,alcohol precipitation concentration as factors,Box-Behnken response surface methodology was used to optimize the extraction process of polysaccharide from Baihe dihuang decoction;and the validation test was conducted.Forty ICR mice were divided into control group,venlafaxine group[positive control,13.5 mg/(kg·d)],Baihe dihuang polysaccharide high-dose and low-dose groups[5.28,2.64 g/(kg·d),by raw material],with 10 mice in each group(half male and half female).Administration groups were given corresponding drug solution intragastrically,and control group was given water 10 mL/kg intragastrically,once a day,for 7 consecutive days.The elevated cross maze test,open field test,forced swimming test and tail suspension test were used to evaluate the effects of the extract prepared by the optimal process on the anxiety-like and depression-like behavior of mice;enzyme-linked immunosorbent assay was used to detect the effects of the extract on the levels of neurotransmitter in cerebral tissueof mice. RESULTS The optimal extraction process of Baihe dihuang decoction was: the water amount of 25 times, extract time of1.5 hours, and alcohol precipitation concentration of 70%. In 3 times of validation test, the average yield and content ofpolysaccharide were 33.10% and 0.62 mg/mg, the relative deviations of which from the predicted values (36.14% and 0.65 mg/mg)were 8.40% and 4.62% respectively (RSD<2%, n=3). The polysaccharide extract of Baihe dihuang decoction could effectivelyincrease the percentages of open-arms entry, the percentages of open-arms time, the total distance of voluntary activities and theactivity distance in central area, and significantly shortened the immobility time of forced

关 键 词：百合地黄汤 多糖 提取工艺 PLACKETT-BURMAN设计 Box-Behnken响应面法 抗抑郁 抗焦虑 小鼠 

分 类 号：R284.2[医药卫生—中药学] R965[医药卫生—中医学]