摘要
目的探讨黄连-乌梅药对与消癌解毒方水煎液中部分生物碱类成分的含量差异,并比较药对与全方对小鼠肠道菌群的影响。方法采用Extend-C18(100mm×2.1mm,1.8μm)色谱柱,以0.1%甲酸水(A)-乙腈(B)进行梯度洗脱,流速0.3mL/min,柱温35℃,进样量2μL;离子化模式为电喷雾离子化(ESI),以正离子模式检测,开展方法学验证和含量测定研究。按照给药不同将小鼠分为正常组、全方高浓度组、全方低浓度组、药对高浓度组和药对低浓度组,连续灌胃给药7d,每组分别收集粪便样本,进行16S rRNA基因测序。结果建立的含量测定方法在一定浓度范围内线性关系良好,方法学验证结果均符合相关要求,木兰花碱与非洲防己碱在黄连、药对以及全方中的含量分别为:(4.433±0.133)(8.905±0.154)mg/g,(3.545±0.033)(9.170±0.051)mg/g,(5.287±0.038)(13.861±0.690)mg/g。全方组和药对组中拟杆菌门Bacteroides的丰度均高于正常组,而全方组和药对组中厚壁菌门Firmicutes的丰度均低于正常组。药对高浓度组中的疣微菌门Verrucomicrobia和变形菌门Proteobacteria的丰度高于其他组。结论液质联用方法和相关参数简便、可靠,可用于有关成分的含量检测。与单味药中含量相比,经过药对配伍后,非洲防己碱含量增加,而木兰花碱含量则略有降低;经过全方配伍后,2种成分的含量均增加。此外,肠道菌群实验表明无论是药对还是全方在给药后,会不同程度地影响肠道菌群内部占比改变,其可能对调节肠道系统平衡,具有一定帮助作用,从而为进一步阐释黄连-乌梅在消癌解毒方中配伍机制提供了参考和依据。
OBJECTIVE To explore the difference of some alkaloids in Coptidis Rhizoma-Mume Fructus decoction and Xiaoai Jiedu Prescription,and compare their effects on the intestinal flora of mice.METHODS The separation was performed on Extend-C18 column(100 mm×2.1 mm,1.8μm)with 0.1% aqueous formic acid(A)-acetonitrile solution(B)as the mobile phase for gradient elution.The flow rate was 0.3 mL/min,column temperature was set at 35℃,and injection volume was 2μL.Electrospray ionization source was applied for the analysis under positive ion model.The method was validated and the samples were determined under this condition.50 mice were divided into ZC group,QFH group,QFL group,YDH group and YDL group according to different dosages.After 7 days of oral administration,fecal samples from each group were collected for16 S rRNA gene sequencing.RESULTS The established detection method showed good linearity within certain concentration ranges and the method validation results meet the relevant requirements.The contents of magnoflorine and columbamine in the single herb,herb pair and formula decoction were(4.433±0.133)and(8.905±0.154)mg/g,(3.545±0.033)and(9.170±0.051)mg/g,(5.287±0.038)and(13.861±0.690)mg/g,respectively.In addition,the abundance of Bacteroides in QF and YD were higher than that in the normal group,but the abundance of Firmicutes was opposite.Interestingly,we found that the abundance of Verrucomicrobia and Proteobacteriain YDH were higher than other groups.CONCLUSION The method and related parameters were simple and reliable,which could be used to detect the content of related components.Compared with single herb,the content of columbamine was increased both in herb pair and Xiaoai Jiedu Prescription;the content of magnoflorine was decreased slightly in herb pair,while increased in Xiaoai Jiedu Prescription.After administering herb decoction,the change of intestinal flora diversity revealed Coptidis Rhizoma-Mume Fructus decoction and Xiaoai Jiedu Prescription could regulate the balance of intestinal system.The
作者
史悦华
范汇森
刘晓
肖平
沈卫星
程海波
孙东东
SHI Yue-hua;FAN Hui-sen;LIU Xiao;XIAO Ping;SHEN Wei-xing;CHENG Hai-bo;SUN Dong-dong(Affiliated Zhong Da Hospital of Southeast University,Nanjing,210009,China;Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine(TCM)Prevention and Treatment of Tumor,Key Laboratory of SATCM for Empirical Formulae Evaluation and Achievements Transformation,Nanjing,210023,China;College of Pharmacy,Nanjing University of Chinese Medicine,Nanjing,210023,China;Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing,Nanjing University of Chinese Medicine,Nanjing,210023,China)
出处
《南京中医药大学学报》
CAS
CSCD
北大核心
2019年第2期199-204,共6页
Journal of Nanjing University of Traditional Chinese Medicine
基金
国家自然科学基金(81673559
81573910
81403079)
江苏省自然科学基金(BK20161045)
江苏高校自然科学基金(16KJB360001)
江苏高校优势学科建设工程资助项目
关键词
黄连-乌梅
配伍
肠道菌群
16S
rRNA基因测序
含量测定
Coptidis Rhizoma-Mume Fructus
compatibility
intestinal flora
16S rRNA gene sequencing
content determination
