中国中药杂志

2020, v.45(16) 3938-3944

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金银花、山银花抗炎药理作用研究
Comparative study on anti-inflammatory effect of Lonicerae Japonicae Flos and Lonicerae Flos

曾安琪;华桦;陈朝荣;刘俐;张敏;罗园;赵军宁;
ZENG An-qi;HUA Hua;CHEN Chao-rong;LIU Li;ZHANG Min;LUO Yuan;ZHAO Jun-ning;Key Research Office for Biological Evaluation of Traditional Chinese Medicine Quality of the State Administration of Chinese Medicine, Sichuan Provincial Traditional Medicine System Development Engineering Technology Research Center, Sichuan Center for Transforming Chinese Medicine, Sichuan Academy of Traditional Chinese Medicine;Chengdu Medical College;Chengdu University of Traditional Chinese Medicine;

摘要(Abstract):

观察金银花提取物和山银花提取物对二甲苯所致小鼠耳肿胀试验中及对脂多糖(LPS)诱导的RAW264.7细胞炎症模型的抗炎作用及作用机制。体内实验采用二甲苯致小鼠耳廓肿胀模型,测定金银花提取物和山银花提取物对二甲苯所致小鼠的耳廓肿胀度及肿胀抑制率;通过苏木精-伊红染色液(HE)染色,观察小数耳廓病理形态变化。体外细胞实验采用LPS诱导的RAW264.7炎症细胞模型,通过细胞活性检测(CCK-8)法检测不同浓度金银花提取物和山银花提取物对RAW264.7细胞的细胞毒作用;采用Griess法检测金银花、山银花提取物对细胞中一氧化氮(NO)分泌量的影响;采用ELISA法检测金银花、山银花提取物对RAW264.7细胞中炎症因子白细胞介素-6(IL-6)、白细胞介素1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的释放量;Western blot法测定金银花、山银花提取物对RAW264.7细胞中环氧酶1(COX1)、COX2以及诱导型一氧化氮合酶(iNOS)蛋白含量的变化。结果显示,金银花提取物和山银花提取物均能显著抑制二甲苯所致小鼠的耳廓肿胀程度,其抑制率与药物剂量呈正相关,且均能减轻小鼠耳组织内淋巴细胞和中性粒细胞的浸润;体外细胞模型中金银花提取物和山银花提取物均可抑制LPS诱导的RAW264.7细胞NO分泌,下调IL-1β,IL-6,TNF-α细胞因子的释放,并下调蛋白iNOS,COX2以及NF-κB p65的含量。金银花提取物和山银花提取物均具有良好的抗炎作用,其作用机制可能与抑制NF-κB信号通路活化有关。
The aim of this paper was to observe the anti-inflammatory action and mechanism of Lonicerae Japonicae Flos extract and Lonicerae Flos extract in xylene-induced ear swelling experiment and lipopolysaccharide(LPS)-induced RAW264.7 cell inflammatory model. In vivo, xylene-induced mouse auricle swelling model was used to detect the auricle swelling degree and swelling inhibition rate of Lonicerae Japonicae Flos extract and Lonicerae Flos extract; the pathological changes of mice auricle were observed by hematoxylin eosin(HE) staining. In vitro, RAW264.7 inflammatory cell model was induced by LPS, where the cytotoxic effects of Lonicerae Japonicae Flos extract and Lonicerae Flos extract on RAW264.7 cells were detected by CCK-8 method; Griess method was used to detect the effect of Lonicerae Japonicae Flos extract and Lonicerae Flos extract on nitric oxide(NO) production, and ELISA method was used to detect the content of inflammatory factors interleukin-6(IL-6), IL-1β, and tumor necrosis factor-α(TNF-α). At last, Western blot was used to detect the protein changes of cyclooxygenase 1(COX1), COX2 and inducible nitric oxide synthetase(iNOS) for RAW264.7 cells. The results showed that both Lonicerae Japonicae Flos extract and Lonicerae Flos extract could significantly inhibit the degree of auricle swelling caused by xylene in mice and the inhibition rate was positively correlated with the drug dose. Furthermore, both of them could reduce the infiltration of lymphocytes and neutrophils in mouse ear tissues. For in vitro experiments, both Lonicerae Japonicae Flos extract and Lonicerae Flos extract inhibited NO secretion in RAW264.7 cells, down-regulated the release of IL-1β, IL-6 and TNF-α, and down-regulated iNOS protein and COX2, NF-κB p65 protein content. In conclusion, both Lonicerae Japonicae Flos extract and Lonicerae Flos extract have good anti-inflammatory effect, and the mechanism may be related with the inhibition of NF-κB signaling pathway.

关键词(KeyWords): 金银花;山银花;抗炎;体内;体外;作用机制
Lonicerae Japonicae Flos;Lonicerae Flos;anti-inflammatory;in vivo;in vitro;mechanism

Abstract:

Keywords:

基金项目(Foundation): 成都市科学技术局产业集群协同创新项目(2016-XT00-00333-GX);; 国家“重大新药创制“科技重大专项(2018ZX09201018-029);; 四川省科技厅重点研发项目(2018SZ0056)

作者(Author): 曾安琪;华桦;陈朝荣;刘俐;张敏;罗园;赵军宁;
ZENG An-qi;HUA Hua;CHEN Chao-rong;LIU Li;ZHANG Min;LUO Yuan;ZHAO Jun-ning;Key Research Office for Biological Evaluation of Traditional Chinese Medicine Quality of the State Administration of Chinese Medicine, Sichuan Provincial Traditional Medicine System Development Engineering Technology Research Center, Sichuan Center for Transforming Chinese Medicine, Sichuan Academy of Traditional Chinese Medicine;Chengdu Medical College;Chengdu University of Traditional Chinese Medicine;

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