赤箭和天麻调节免疫功能和肠道菌群作用比较  

Comparison of Effect of Chijian and Gastrodiae Rhizoma on Regulating Immune Function and Intestinal Microflora

作　　者：席蕴文 康利平[2] 孙建辉[3] 程铭恩 李洪梅[3] 黄璐琦 XI Yun-wen;KANG Li-ping;SUN Jian-hui;CHENG Ming-en;LI Hong-mei;HUANG Lu-qi(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;National Resource Center for Chinese Meteria Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012,China)

机构地区：[1]南京中医药大学药学院,南京210023 [2]中国中医科学院中药资源中心,北京100700 [3]中国中医科学院中药研究所,北京100700 [4]安徽中医药大学药学院,合肥230012

出　　处：《中国实验方剂学杂志》2021年第12期117-124,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中央本级重大增减支项目(2060302)。

摘　　要：目的:对赤箭(天麻地上部分)和天麻中腺苷、天麻素、对羟基苯甲醇、对羟基苯甲醛、巴利森苷B和A的成分含量,以及赤箭水提液和天麻水提液调节免疫功能和肠道菌群作用进行比较,评价赤箭进一步研究和开发的可行性。方法:采用超高效液相色谱法(UPLC)测定6种成分的含量,流动相0.1%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~4 min,0.5%B;4~5 min,0.5%~2%B;5~10 min,2%~15%B;10~12 min,15%~20%B;12~15 min,20%~95%B;15~17 min,95%B;17~17.5 min,95%~0.5%B;17.5~20 min,0.5%B),流速0.5 mL·min^(-1),检测波长270 nm。采用环磷酰胺致小鼠免疫功能低下模型,测定廓清指数、校正廓清指数及外周血象,采用体外淋巴细胞转化试验测定B淋巴细胞增殖,基于16S r DNA技术测定肠道菌群并进行生物信息学分析,对赤箭水提液和天麻水提液进行药理活性比较。结果:赤箭粉末、醇提液中6个成分总量分别高于天麻粉末,醇提液,赤箭水提液和天麻水提液中6个成分含量总和相近,且赤箭粉末和天麻粉末中天麻素和对羟基苯甲醇总量均符合2020年版《中华人民共和国药典》的相关要求。与空白组比较,赤箭水提液中剂量(10 g·kg^(-1))组和天麻水提液中、低剂量(10,5 g·kg^(-1))组可明显增加免疫功能低下小鼠的廓清指数(P<0.05);赤箭水提液、天麻水提液高剂量组(20 g·kg^(-1))均可明显增高外周血中红细胞和红细胞压积水平(P<0.05,P<0.01);400 g·L^(-1)天麻水提液和100 g·L^(-1)赤箭水提液对脂多糖诱导的B淋巴细胞增殖具有促进作用。肠道菌群研究显示,与空白组比较,在门水平上,赤箭水提液和天麻水提液增加拟杆菌门的相对丰度,降低厚壁菌门的相对丰度;在属水平上,赤箭水提液、天麻水提液均可增加PrevotellaceaeUCG-001和RuminococcaceaeUCG-005的相对丰度,降低厌氧棍状菌属,unclassifiedfErysipelotrichaceae和CandidatusStoquefichus的相对丰度,这些肠道菌与免疫系统、细胞增�Objective:To compare the contents of adenosine,gastrodin,p-hydroxybenzyl alcohol,p-hydroxybenzaldehyde,parisinin B and parisinin A in Chijian (the aerial part of Gastrodia elata) and Gastrodiae Rhizoma,and compare their effects on immune function and intestinal microflora,evaluating whether it is necessary to study and develop Chijian.Method:The contents of these six constituents were determined by ultra performance liquid chromatography(UPLC),the mobile phase was 0.1%formic acid aqueous solution(A)-acetonitrile(B)for gradient elution(0-4 min,0.5%B;4-5 min,0.5%-2%B;5-10 min,2%-15%B;10-12 min,15%-20%B;12-15 min,20%-95%B;15-17 min,95%B;17-17.5 min,95%-0.5%B;17.5-20 min,0.5%B),the flow rate was 0.5 mL·min^(-1),the detection wavelength was 270 nm.The difference of pharmacological activity of water extracts of Chijian and Gastrodiae Rhizoma was compared,the clearance index,corrected clearance index and peripheral blood were measured in mice model with low immune function induced by cyclophosphamide,B lymphocyte proliferation was determined by lymphocyte transformation test in vitro,intestinal microflora was analyzed by 16S r DNA technology and bioinformatics was conducted.Result:The total contents of these six components in powder and ethanol extract of Chijian were higher than that of Gastrodiae Rhizoma,but the total contents of these six components in their water extract were similar,and the total contents of gastrodin and p-hydroxybenzyl alcohol met the requirements of the 2020 edition of Chinese Pharmacopoeia.Compared with the blank group,the clearance index of immunocompromised mice was significantly increased in the middle-dose(10 g·kg^(-1))group of Chijian water extract,middle-and low-dose(10,5 g·kg^(-1))groups of Gastrodiae Rhizoma water extract(P<0.05),the levels of erythrocyte and hematocrit in peripheral blood were significantly increased in the high-dose(20 g·kg^(-1))groups of water extracts of Chijian and Gastrodiae Rhizoma(P<0.05,P<0.01),water extract of Gastrodiae Rhizoma with concentration of 400 g

关 键 词：天麻 赤箭 脂多糖(LPS) 超高效液相色谱法(UPLC) 免疫功能 肠道菌群 化学成分 

分 类 号：R378.2[医药卫生—病原生物学] R392[医药卫生—基础医学]