野菊花外用治疗婴幼儿急性湿疹的药效学及免疫机制初探

发布时间：2018-06-18 11:35

本文选题：野菊花 + 湿疹　；参考：《成都中医药大学》2016年硕士论文

【摘要】：研究目的：观察野菊花不同提取物对于湿疹小鼠皮肤的干预作用,初探野菊花药物治疗湿疹的药效及相关免疫机制。研究方法：将56只清洁级KM小鼠随机平均分为空白组、模型组、阳性对照组(儿肤康搽剂组)、野菊花水提物组、95%、70%及50%浓度乙醇提物组,每组8只,采用2,4一二硝基氯苯(DNCB)腹部致敏,右耳部激发方法建立湿疹小鼠模型,同时于致敏第一天(d1)开始给予各组相应药物外用。末次激发后24h(d12)处死小鼠,收集小鼠血清及右耳耳片组织,测量小鼠右耳耳片肿胀度并在光镜下观察组织病理变化：采用高效液相色谱指纹图谱(HPLC)建立野菊花四种提取物指纹图谱,最后对空白组、模型组、阳性对照组及野菊花四组提取物中两药效及化合物成分优异的野菊花水提物组和70%醇提取物组采用酶联免疫吸附测定(Elisa)检测小鼠血清免疫球蛋白E (IgE)水平,免疫组化检测小鼠激发耳廓组织中白细胞介素-4(IL-4)、干扰素γ(IFN-γ)平均光密度值。研究结果：1、与空白组相较,模型组右耳部激发处出现明显红斑,水肿,少数皮肤可见糜烂、渗出结痂,同时光镜下符合湿疹病理表现；组织切片肿胀度明显增加,具有显著差异(P0.01),阳性对照组(儿肤康)与野菊花四种提取物组小鼠耳片肿胀值结果均小于模型组,其结果具有显著的统计学差异(P0.01),各组均能不同程度地改善激发右耳组织肉眼及病理学改变；野菊花水提取物组优于野菊花不同比例醇提取物(P0.01),与阳性药物疗效作用相当(P0.05)；2、成功建立野菊花提取物高效液相指纹图谱,图谱显示野菊花水提物主成分相对于醇提取物更为丰富,两者内均具有特殊波峰呈现,70%组醇提取物在各醇提物主峰表现中波峰最为全面；3、与空白组相较,湿疹模型小鼠激发右耳组织IL-4平均光密度值明显升高(P0.01),IFN-γ水平表达明显降低(P0.01),模型血中IgE水平明显升高(P0.01)；较模型组而言野菊花水提物可降低小鼠血清IgE水平(P0.05),显著降低皮损中IL-4因子的表达(P0.01),同时能够上调IFN-γ含量(P0.05)；而野菊花70%醇提取物组较模型组而言能显著升高IFN-γ(P0.01),下调小鼠湿疹皮损中的IL-4水平(P0.05),具有降低血清IgE的趋势(P0.05)。研究结论：1、野菊花药物具有治疗湿疹的作用,有效成分可能多存在于水提物及70%醇提取物的主要组分之中；2、野菊花能通过免疫调节改善湿疹小鼠TH1/TH2免疫功能失衡现象,其机制与抑制IL-4表达及提升IFN-γ的分泌相关,不同提取物间存在着一定的作用差异；水提物能有效抑制IgE水平。
[Abstract]:Objective: To observe the effect of different extracts of chrysanthemum chrysanthemum on the skin of eczema in mice and explore the efficacy and immune mechanism of Chrysanthemum in the treatment of eczema. Methods: 56 clean KM mice were randomly divided into blank group, model group, positive control group (ER Fukang liniment group), water extracts of Chrysanthemum chrysanthemum, 95%, 70% and 50%. In the ethanol extract group, 8 rats in each group were treated with 2,4 one or two nitrochlorobenzene (DNCB) abdominal sensitization, the right ear excitation method was used to establish the eczema mouse model. At the same time, the first day (D1) was used to give the corresponding drug external use. After the last excitation, 24h (D12) was executed in mice, the mice were collected and the right ear ear slices were collected to measure the swelling degree of the right ear ear slices in mice. The histopathological changes were observed under light microscope: the fingerprint of four extracts of chrysanthemum Chrysanthemi was established by high performance liquid chromatography (HPLC). Finally, the enzyme linked immunosorbent assay was used in the blank group, model group, positive control group and four group of chrysanthemum chrysanthemum extract, which had two effective and excellent compounds in chrysanthemum and 70% alcohol extract group. The level of serum immunoglobulin E (IgE) in mice was detected by adsorption (Elisa). The average optical density of interleukin -4 (IL-4) and interferon gamma (IFN- gamma) in the auricle tissue of mice was detected by immunohistochemistry. The results were as follows: 1, compared with the blank group, there was obvious erythema, edema, erosion and crusting in the right ear of the model group. At the same time, the pathological features of eczema were conformed to the pathological features of the eczema, and the swelling degree of tissue sections increased significantly (P0.01). The results of the ear swelling of the four extracts of the positive control group (ER Fukang) and the chrysanthemum chrysanthemum were less than the model group. The results showed significant statistical difference (P0.01). All the groups could improve the excite right ear in varying degrees. The water extract group of chrysanthemum chrysanthemum was better than the alcohol extract of wild chrysanthemum (P0.01), and the effect was similar to that of the positive drug (P0.05). 2, the HPLC fingerprint of the extract of chrysanthemum chrysanthemum was successfully established. The map showed that the main component of the aqueous extract of Chrysanthemum Chrysanthemum was more abundant than the alcohol extract. Both of them had special characteristics. The peak of the 70% groups of alcohol extracts in the main peak of each alcohol extract was the most comprehensive. 3, compared with the blank group, the average light density of IL-4 in the right ear tissue of the eczema model mice increased significantly (P0.01), the expression of IFN- gamma decreased significantly (P0.01), and the level of IgE in the model blood was significantly increased (P0.01); compared with the model group, the water extraction of wild chrysanthemum was more than that of the model group. The serum IgE level (P0.05) was reduced and the expression of IL-4 factor in skin lesions (P0.01) was significantly reduced, and the content of IFN- gamma (P0.05) was up-regulated, while the 70% alcohol extract group of Chrysanthemum Chrysanthemum could significantly increase IFN- gamma (P0.01) and reduce the IL-4 level (P0.05) in the skin lesions of mice (P0.05), and had a tendency to reduce IgE in the serum (P0.05). Conclusion: 1, the drug of chrysanthemum chrysanthemum has the effect of treating eczema. The effective components may exist in the main components of water extract and 70% alcohol extract. 2, wild chrysanthemum can improve the immune function imbalance of TH1/TH2 in the eczema mice by immunoregulation. The mechanism is related to the inhibition of the expression of IL-4 and the secretion of IFN- gamma. Water extracts can effectively inhibit the level of IgE.
【学位授予单位】：成都中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R275.9

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