目的 探究青翘和老翘抗炎活性差异的质量标志物(quality marker)。方法 采用高效液相色谱(HPLC)结合化学计量学对青翘和老翘进行指纹图谱分析;采用脂多糖(LPS)诱导小鼠单核巨噬细胞RAW264.7炎症模型,以炎症因子一氧化氮(NO)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的水平,评价青翘和老翘抗炎活性差异;采用SPSS软件关联指纹图谱色谱峰与抗炎活性的关系,考察青翘和老翘的质量差异性成分和抗炎活性之间的相关性;采用分子对接技术验证质量标质物与炎症核心靶点的结合能力。结果 青翘和老翘指纹图谱中分别被标定14个共有峰和16个共有峰,10个色谱峰(VIP>1)被筛选为青翘和老翘质量差异性成分;青翘、老翘均有明显的抗炎活性,抑制NO的效果,老翘>青翘(P<0.001),抑制TNF-α和IL-6方面,青翘>老翘(P<0.001)。谱效关联分析显示,造成青翘和老翘抗炎活性差异的标志物有10个。依据质量标志物“五原则”优选连翘苷、连翘酯苷A、芦丁和连翘酯素4个成分,作为青翘和老翘抗炎差异的质量标志物。分子对接的结合能(≤-5)表明优选成分与抗炎核心靶点间具有良好的结合能力。结论 本研究建立的HPLC指纹图谱方法良好可行,筛选出4个成分为青翘和老翘抗炎活性差异的质量标志物,为青翘和老翘科学应用提供更充分的理论依据。
Abstract
OBJECTIVE To explore the quality markers of anti-inflammatory activity difference between Qingqiao and Laoqiao. METHODS High performance liquid chromatography (HPLC) combined with stoichiometry was used to analyze the fingerprints of Qingqiao and Laoqiao. The inflammatory model of mouse monocyte macrophage RAW264.7 was induced by lipopolysaccharide (LPS). The levels of inflammatory factors nitric oxide (NO), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were used to evaluate the difference of anti-inflammatory activity between Qingqiao and Laoqiao.SPSS software was used to correlate the relationship between fingerprint peaks and anti-inflammatory activity, and the correlation between the quality difference components and anti-inflammatory activity of Qingqiao and Laoqiao was investigated.Molecular docking technology was used to verify the binding ability of quality marker to core targets of inflammation. RESULTS Fourteen common peaks and 16 common peaks were identified in the fingerprints of Qingqiao and Laoqiao, respectively. Ten chromatographic peaks (VIP>1) were selected as the quality difference components of Qingqiao and Laoqiao.Both Qingqiao and Laoqiao had obvious anti-inflammatory activity and inhibited NO. Laoqiao>Qingqiao (P<0.001), in terms of inhibiting TNF-α and IL-6, Qingqiao>Laoqiao (P<0.001).Spectrum-effect correlation analysis showed that there were 10 markers that caused the difference in anti-inflammatory activity between Qingqiao and Laoqiao.According to the "five principles" of quality markers, the four components of forsythin, forsythoside A, rutin and phillygenin were selected as the quality markers for the difference of anti-inflammatory between Qingqiao and Laoqiao. The binding energy of molecular docking (≤-5) indicated that the preferred components had good binding ability to the anti-inflammatory core target. CONCLUSION The HPLC fingerprint method is established in this study, which is good and feasible. Four components were selected as quality markers for the difference in anti-inflammatory activity between Qingqiao and Laoqiao, providing a more sufficient theoretical basis for the scientific application of Qingqiao and Laoqiao.
关键词
青翘 /
老翘 /
指纹图谱 /
抗炎活性 /
谱-效相关 /
质量标志物
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Key words
Qingqiao /
Laoqiao /
fingerprint /
anti-inflammatory activity /
spectrum-effect correlation /
quality marker
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脚注
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基金
“十三五”国家重点研发计划项目子课题资助(2017YFD0600702-3);河南大学本科生开放性课题项目资助(20223101306)
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