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 610041, China.
Chengdu Medical College Chengdu 610083, China.
Zhongguo Zhong Yao Za Zhi. 2020 Aug;45(16):3938-3944. doi: 10.19540/j.cnki.cjcmm.20200520.401.
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.
本文旨在通过二甲苯致耳肿胀实验以及脂多糖(LPS)诱导的RAW264.7细胞炎症模型,观察金银花提取物和忍冬花提取物的抗炎作用及机制。在体内实验中,采用二甲苯致小鼠耳廓肿胀模型,检测金银花提取物和忍冬花提取物对耳廓肿胀程度及肿胀抑制率;通过苏木精-伊红(HE)染色观察小鼠耳廓的病理变化。在体外实验中,用LPS诱导RAW264.7炎症细胞模型,采用CCK-8法检测金银花提取物和忍冬花提取物对RAW264.7细胞的细胞毒性作用;采用Griess法检测金银花提取物和忍冬花提取物对一氧化氮(NO)生成的影响,采用ELISA法检测炎症因子白细胞介素-6(IL-6)、IL-1β和肿瘤坏死因子-α(TNF-α)的含量。最后,运用蛋白质免疫印迹法检测RAW264.7细胞中环氧合酶1(COX1)、COX2和诱导型一氧化氮合酶(iNOS)的蛋白变化。结果表明,金银花提取物和忍冬花提取物均能显著抑制二甲苯所致的小鼠耳廓肿胀程度,且抑制率与药物剂量呈正相关。此外,二者均可减少小鼠耳部组织中淋巴细胞和中性粒细胞的浸润。对于体外实验,金银花提取物和忍冬花提取物均能抑制RAW264.7细胞中NO的分泌,下调IL-1β、IL-6和TNF-α的释放,并下调iNOS蛋白以及COX2、NF-κB p65蛋白含量。综上所述,金银花提取物和忍冬花提取物均具有良好的抗炎作用,其机制可能与抑制NF-κB信号通路有关。