Anti-inflammatory effect of Ganluyin, a Chinese classic prescription, in chronic pharyngitis rat model
Anti-inflammatory effect of Ganluyin, a Chinese classic prescription, in chronic pharyngitis rat model
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甘露饮对慢性咽炎大鼠模型的抗炎作用
DOI：
10.1186/s12906-020-03057-5
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发表时间：
2020-08-28
影响因子：
3.9
通讯作者：
Chen, Su-Hong
中科院分区：
文献类型：
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作者：
Chen, Ye-Hui;Luo, Rong;Chen, Su-Hong
背景甘露饮是一个著名的经典方剂，在我国许多地区用于治疗慢性咽炎等炎症性疾病。然而，它尚未被开发为现代药物，其抗炎机制仍不清楚。本研究的目的是评估GLY的抗炎疗效和潜在的机制在大鼠模型CP。MethodsGLY的化学配置文件进行了分析，通过HPLC-UV。我们使用了小鼠耳水肿模型和大鼠足水肿模型。具体地，使用二甲苯在小鼠的一只耳朵的表面上诱导水肿，并且将角叉菜胶皮下注射到大鼠的右后爪中以诱导爪水肿。测量并记录爪厚度、耳重量和耳灌注。采用5%氨水刺激大鼠咽喉部建立慢性前列腺炎模型，观察甘利宁对慢性前列腺炎的治疗作用。ELISA法检测血清中白细胞介素6（IL-6）、白细胞介素1β（IL-1β）、肿瘤坏死因子α（TNF-α）、前列腺素E2（PGE 2）水平，免疫组化法和Western blot法检测咽喉部环氧化酶2（考克斯-2）和核因子κ-B p65（NF-κB p65）蛋白表达，评价甘利咽的抗炎作用机制。结果从甘草中鉴定出4种黄酮类化合物：柚皮苷、新橙皮苷、黄芩苷和汉黄芩苷。GLY在6.2和12.4g/kg的剂量下对二甲苯诱导的小鼠耳肿胀和耳血流量表现出显著的抑制作用，并且显著改善大鼠右后足肿胀。机制研究发现，GLY的抗炎活性与抑制促炎细胞因子如IL-1β、IL-6、TNF-α和PGE 2有关，并且GLY降低咽喉部考克斯-2和NF-κB p65蛋白的表达，减轻咽喉损伤，减少炎性渗出物。血液学分析表明，用GLY治疗防止了白色血细胞（WBC）、中性粒细胞（NEUT）、淋巴细胞（LYMPH）和单核细胞（MONO）水平的增加。降低淋巴结和单核细胞水平，降低考克斯-2和NF-κB p65蛋白的表达。这些发现可能为进一步研究GLY作为治疗炎症性疾病（如CP）的合适候选药物奠定基础。
BackgroundGanluyin (GLY) is a famous classical prescription with a long history of use as a treatment for inflammatory conditions such as chronic pharyngitis (CP) in many parts of China. However, it has not been developed as a modern pharmaceutic and its anti-inflammatory mechanisms remain unclear. The aim of this study was to assess the anti-inflammatory efficacy of GLY and potential mechanisms in a rat model of CP.MethodsThe chemical profile of GLY was analyzed by HPLC-UV. We used a mouse model of ear edema and a rat model of paw edema. Specifically, xylene was used to induce edema on the surface of one ear in mice, and carrageenan was injected subcutaneously into the right hind paws of rats to induce paw edema. The paw thickness, ear weight, and ear perfusion were measured and recorded. The CP model in rats was induced by irritating the throat with 5% ammonia and was used to evaluate the therapeutic efficacy of GLY. Levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor (TNF-α), and prostaglandin E2 (PGE2) were measured by ELISA in serum, and protein expression of cyclooxygenase-2 (COX-2) and nuclear factor kappa-B p65 (NF-κB p65) in the throat were detected by immunohistochemistry and Western blot to evaluate the anti-inflammatory mechanism of GLY. Hematological assays were also conducted.ResultsThere were four flavonoids identified in GLY: naringin, neohesperidin, baicalin, and wogonoside. The oral administration of GLY showed a significant inhibitory effect on xylene-induced ear swelling and ear blood flow in mice and significantly ameliorated rat right hind paw edema at doses of 6.2 and 12.4 g/kg. Mechanistic studies found that the anti-inflammatory activity of GLY was related to the inhibition of pro-inflammatory cytokines such as IL-1β, IL-6, TNF-α, and PGE2 and that GLY reduced the expression of COX-2 and NF-κB p65 proteins in the throat, attenuated throat injury, and reduced inflammatory exudates. Hematological analysis showed that treatment with GLY prevented increases in white blood cell (WBC), neutrophil (NEUT), lymphocyte (LYMPH) and monocyte (MONO) levels.ConclusionsThese studies indicated that GLY has beneficial anti-inflammatory effects on CP and that it acts through reducing pro-inflammatory factors such as IL-1β, IL-6, TNF-α, and PGE2, as well as decreasing WBC, NEUT, LYMPH and MONO levels and decreasing the expression of COX-2 and NF-κB p65 proteins. These findings may lay the groundwork for further studies of GLY as a suitable candidate for the treatment of inflammatory diseases such as CP.