基于化学成分与肠道菌群探讨消癌解毒方中黄连-乌梅药对的配伍机制
The Study of Compatibility Mechanism of Coptidis Rhizoma-Mume Fructus in Xiaoai Jiedu Prescription from Perspective of Chemical Components and Intestinal Microflora
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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(100 mm×2.1 mm,1.8 μm)色谱柱,以0.1%甲酸水(A)-乙腈(B)进行梯度洗脱,流速0.3 mL/min,柱温35 ℃,进样量2 μL;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7 d,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。
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关键词:
- 黄连-乌梅 /
- 配伍 /
- 肠道菌群 /
- 16S rRNA基因测序 /
- 含量测定
Abstract: 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, 1.8 μm) 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 2 μL. 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 for 16S 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 Firmicuteswas opposite. Interestingly, we found that the abundance of Verrucomicrobia and Proteobacteria in 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 results further explain the compatibility mechanism of the two drugs in this prescription. -
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