本研究采用脂多糖(lipopolysaccharide，LPS)诱导大鼠全身感染性炎症模型，通过苏木精-伊红(hematoxylin eosin，H&E)染色观察模型大鼠肺和结肠组织病理改变，并以炎症因子一氧化氮(nitric oxide，NO)、白细胞介素-6(interleukin-6，IL-6)和肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)为指标，评价裸花紫珠水提物干浸膏的抗炎作用。实验按照广东药科大学实验动物福利伦理委员会指导政策严格执行(批准号:gdpulac2022132)。通过网络药理学预测裸花紫珠提取物中具有抗炎作用的化学成分和潜在作用靶点，甄别质量标志物，并采用高效液相色谱(high performance liquid chromatography，HPLC)法测定它们的含量。结果表明，裸花紫珠水提物干浸膏可显著减少炎症大鼠体TNF-α和IL-6的释放，降低NO水平(P＜0。01)。裸花紫珠中83个化学成分通过调控磷脂酰肌醇 3激酶-蛋白激酶(phosphatidylinositol-3-hydroxykinase-protein kinase，PI3K-Akt)信号通路、酪氨酸激酶-信号转导因子和转录激活因子 3(Janus kinase-signal transducer and activator of transcription 3，JAK-STAT3)信号通路等发挥抗炎作用。结合质量标志物特有性、有效性、可测性以及量值传递规律等要素筛选得到6-羟基木犀草苷、木犀草苷、咖啡酸、毛蕊花糖苷、异毛蕊花糖苷和连翘酯苷B可能为裸花紫珠提取物的抗炎活性成分，通过LPS诱导小鼠单核巨噬细胞RAW264。7炎症模型验证了它们的抗炎活性。最后，以这6个化合物作为含量测定指标成分，评价了 16批海南产裸花紫珠药材的质量。综上，本研究通过体内外实验证实了裸花紫珠水提物干浸膏的抗炎作用，6-羟基木犀草苷、木犀草苷、咖啡酸、毛蕊花糖苷、异毛蕊花糖苷和连翘酯苷B是裸花紫珠抗炎活性成分，可用于裸花紫珠药材及系列制剂的质量评价。
Quality evaluation of Callicarpa nudiflora from Hainan Province based on simultaneous determination of six anti-inflammatory active components by HPLC
The anti-inflammatory efficacy of Callicarpa nudiflora extract were evaluated upon lipopolysaccha-ride(LPS)-induced infective inflammation in rats.Pathological changes of lung and colon tissues observed with hematoxylin-eosin(H&E)staining,together with inflammatory factors in serum,including nitric oxide(NO),inter-leukin-6(IL-6)and tumor necrosis factor-α(TNF-α),were applied to assess the mitigating effects of C.nudiflora water extract on model rats.The experimental protocol strictly adhered to the guidelines of the Ethics Committee of Animal Research of Guangdong Pharmaceutical University(Approval:gdpulac2022132).Quality markers with anti-inflammatory activities were recognized by network pharmacology analysis.Subsequently,a reliable method for simultaneously quantifying six components was established.As a result,the pathological injury on lung and colon tissues of model rats induced by LPS were improved by C.nudiflora water extract.Compared with model group,C.nudiflora extract had decreased the release of proinflammatory factors in serum,including TNF-α and IL-6,and reduced the NO(P＜0.01).Network pharmacological analysis obtained 83 chemical components,466 component targets,584 disease targets and 129 action targets,which were mainly concentrated in 624 biological processes such as inflammatory response and positive regulation of nitric oxide biosynthesis,as well as 162 pathways such as phosphatidylinositol-3-hydroxykinase-protein kinase(PI3K-Akt)signal pathway and Janus kinase-signal transducer and activator of transcription 3(JAK-STAT3)signal pathway.Upon analyzing their specificity,effectiveness,detectability,and quantity transfer rule from herb to extract,6-hydroxyluteolin 7-glucoside,cynaroside,caffeic acid,acteoside,isoacteoside,and forsythoaside B were identified as the quality markers.Their anti-inflammatory activi-ties were verified with LPS-induced inflammation model of RAW264.7 cells.Then contents of these 6 quality markers in 16 batches of C.nudiflora were determined.Thereby,the anti-inflammatory efficacy of C.nudiflora extract were confirmed in vivo,6-hydroxyluteolin 7-glucoside,cynaroside,caffeic acid,acteoside,isoacteoside,and forsythoaside B were identified as anti-inflammatory ingredients,which can be used as quality control indicators for the herb and related formulation.
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