痛风性关节炎湿热证病证结合动物模型研究

发布时间：2018-05-04 14:56

本文选题：痛风性关节炎 + 湿热证　；参考：《湖南中医药大学》2012年硕士论文

【摘要】：目的：通过多因素复合造模的方法,以当前广泛应用的湿热证大鼠模型和经典的痛风性关节炎模型作为对照,探讨建立中医痛风性关节炎湿热证大鼠模型的方法,旨在开展痛风性关节炎湿热证病证结合动物模型研究,为病证结合治疗的临床疗效机理研究提供新的思路和实验基础。方法：1、饮食高脂高糖饮食：普通饲料加用蜂蜜水自由饮用,且按大鼠体质量,灌服油脂和白酒隔日交替。 2、分组选用健康成年SD大鼠40只,按随机法将大鼠分为4组：空白对照组(A组),痛风对照组(B组),湿热对照组(C组),痛风加湿热模型组(D组),每组10只。 3、造模在高脂高糖饮食、湿热环境的综合因素作用下,建立湿热证痛风性关节炎动物模型：①正常对照组：在恒温、恒湿环境下,蒸馏水自由饮水,普通饲料喂养；②痛风对照组：普通饲料+蒸馏水自由饮水+关节腔注射药物；③湿热对照组：高脂高糖饮食+湿热环境；④痛风加湿热模型组：高脂高糖饮食+湿热环境+关节腔注射药物。 4、观察及指标检测对比观察实验大鼠造模前后的一般状态、饮食量、饮水量、体重、大便性状等的变化。用Bradford法检测大鼠尿液中的水通道蛋白2(AQP2),放射免疫法检测内皮素(ET)及降钙素基因相关肽(CGRP)。鼠处死后,迅速取出踝关节滑膜组织、舌组织,置于10%甲醛溶液中固定,石蜡包埋制成切片,HE染色,光镜下观察。结果：内皮素(ET)、降钙素基因相关肽(CGRP)、AQP2作为湿热证的量化指标,血尿酸、步态、关节组织形态学变化作为痛风性关节炎的指标。A组各项指标为正常值(对照用),B组比较出现步态改变、关节滑膜组织形态学改变,C组出现内皮素升高、降钙素基因相关肽降低、尿AQP2含量降低,D组检测结果与B、C两组比较如果同时出现B、C两组的改变(血尿酸,步态、关节组织形态学尿AQP2含量),提示该模型制作成功。结论：实验结果显示,在高脂高糖饮食+湿热环境+关节腔注射药物的综合作用下,可以成功复制痛风性关节炎湿热证的动物模型。此方法是对“病证结合”的痛风性关节炎湿热证动物模型实验研究的积极探索,将对病证结合治疗的临床疗效机理研究提供新的思路和实验基础。
[Abstract]:Objective: to establish a rat model of damp heat syndrome of gouty arthritis of traditional Chinese medicine by means of multi factor compound model making and the classic gouty arthritis model which is widely used as a contrast. The study of the mechanism of clinical efficacy provides new ideas and experimental basis.
Methods: 1. Diet high fat and high sugar diet: regular feed plus honey water for free consumption.
2, 40 healthy adult SD rats were selected, and the rats were divided into 4 groups according to random method: blank control group (group A), gout control group (group B), damp heat control group (group C), gout and damp heat model group (group D), 10 rats in each group.
3, under the action of high fat and high sugar diet and hot and humid environment, the model of damp heat syndrome of gouty arthritis was established. (1) normal control group: under constant temperature, constant humidity, free drinking water of distilled water, feeding of ordinary feed; 2. Gout control group: ordinary feed + steam distilled water free drinking water + joint injection medicine; damp heat pair Photo group: high fat and high sugar diet + damp heat environment; 4. Gout plus damp heat model group: high fat and high sugar diet + damp heat environment + joint cavity injection drugs.
4, observation and index detection were observed and compared to observe the general state of the rats before and after the experimental model, the amount of diet, the amount of drinking water, the weight and the stool character. The Bradford method was used to detect the aquaporin 2 (AQP2) in the urine of the rats, and the radioimmunoassay was used to detect the endothelin (ET) and calcitonin gene related peptide (CGRP). After the rat was killed, the synovial membrane group of the ankle joint was quickly removed. The tissues of tongue and tongue were fixed in 10% Formaldehyde Solution, and paraffin embedded into slices, stained with HE and observed under light microscope.
Results: endothelin (ET), calcitonin gene related peptide (CGRP), AQP2 as a quantitative index of damp heat syndrome, blood uric acid, gait and joint histomorphological changes as indexes of group.A for gouty arthritis were normal values (control). In group B, gait changes, histomorphological changes of synovial synovium in joint, and elevation of endothelin in group C were decreased. Calcitonin gene related peptide decreased, urine AQP2 content decreased, D group detection results compared with B, C two group if B, C two group changes (blood uric acid, gait, joint histomorphological AQP2 content), suggesting that the model was made successfully.
Conclusion: the experimental results show that the animal model of hygrothermal syndrome of gouty arthritis can be successfully replicated under the combined effect of high fat and high sugar diet + humid heat environment plus joint cavity injection. This method is an active exploration of the animal model of damp heat syndrome of gouty arthritis with the combination of disease and syndrome. The study of bed mechanism provides new ideas and experimental basis.

【学位授予单位】：湖南中医药大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R-332

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