麝香乌龙丸对AA大鼠垂体组织形态及ACTH、β-EP表达的影响

发布时间：2018-07-17 02:34

【摘要】：目的通过观察麝香乌龙丸对大鼠佐剂性关节炎（Adjuvant Arthritis, AA）模型垂体组织形态及促肾上腺皮质激素（adrenocorticotropic hormone, ACTH）、β-内啡肽（β-endorphin, β-EP）表达的影响，探讨麝香乌龙丸治疗类风湿关节炎（Rheumatoid arthritis, RA）的镇痛作用机制，为其临床应用提供实验与理论依据。方法将60只雄性Wistar大鼠适应性喂养7d后，随机抽取12只，作为正常组。除正常组外，其余48只大鼠均使用乙醚麻醉，于右后足跖注射完全弗氏佐剂（Freund’s complete adjuvant, FCA），建立AA大鼠模型，造模第15天按关节炎指数（arthritis index, AI）随机分成模型组、麝香乌龙丸低剂量组、麝香乌龙丸中剂量组、麝香乌龙丸高剂量组，每组12只。分别给予麝香乌龙丸低、中、高剂量组麝香乌龙丸0.3g·kg-1，0.6g·kg-1，1.2g·kg-1（生理盐水混悬，分别相当于成人用药剂量的3，6，12倍），正常组与模型组给予灌胃等容量生理盐水，每日1次，连续给药2周。第15天将所有大鼠处死后打开脑腔，取出垂体，垂体组织形态观察采用苏木素—伊红（HE）染色方法，在光镜下观察各组垂体组织形态的变化，，ACTH、β-EP的表达变化采用免疫组化法进行观察，统计阳性细胞表达率。结果1对大鼠一般情况的影响：正常组大鼠饮食正常，皮毛光亮，精神状态佳。其余大鼠注射FCA后出现注射足肿胀并出现左后足及前足红肿，活动减少，饮食饮水减少，毛发稀疏、无光泽，行走困难。给药期间，麝香乌龙丸治疗组大鼠较模型组一般情况较好，反应灵敏，活动增多，足踝和足掌肿胀逐渐减轻，皮毛有光泽。2对大鼠AI的影响：与正常组相比造模各组大鼠AI显著升高（P＜0.05）。治疗后与模型组比较，麝香乌龙丸低、中、高各组AI均明显下降（P＜0.05），尤以麝香乌龙丸高剂量组下降明显。3对大鼠垂体组织形态的影响：光镜下观察各组HE染色后切片发现，与正常组相比，其余各组垂体组织形态均无明显改变。4对垂体ACTH表达的影响：采用免疫组化实验方法观察各组大鼠垂体组织内ACTH表达变化，结果各组ACTH细胞阳性率分别为正常组40.23±3.43，模型组56.82±3.97，麝香乌龙丸低剂量组53.14±4.16，麝香乌龙丸中剂量组50.14±3.39，麝香乌龙丸高剂量组46.87±3.31。与正常组相比，其余4组ACTH细胞阳性表达均明显升高，且差异具有统计学意义（P＜0.01）。麝香乌龙丸低剂量组与模型组ACTH阳性表达无显著性差异（P＞0.05）。麝香乌龙丸中剂量组与麝香乌龙丸高剂量组ACTH细胞阳性表达明显低于模型组，差异具有统计学意义（P＜0.05或P＜0.01），麝香乌龙丸低、中、高三组对垂体ACTH阳性细胞表达的影响有剂量相关性，其中以麝香乌龙丸高剂量组下调ACTH表达最为明显。5对垂体β-EP表达的影响：采用免疫组化实验方法观察各组大鼠垂体组织内β-EP表达变化，发现各组β-EP细胞阳性率分别为正常组30.84±2.19，模型组39.72±2.66，麝香乌龙丸低剂量组43.55±3.59，麝香乌龙丸中剂量组46.69±3.62，麝香乌龙丸高剂量组49.47±3.17。相较于正常组，模型组与麝香乌龙丸低、中、高各组β-EP细胞阳性表达均明显升高，且差异具有统计学意义（P＜0.01）。麝香乌龙丸低、中、高剂量组β-EP阳性表达均明显高于模型组，且差异具有统计学意义（P＜0.01），麝香乌龙丸低、中、高三组对垂体β-EP阳性细胞表达的影响有剂量相关性，其中以麝香乌龙丸高剂量组升高β-EP表达最为明显。结论麝香乌龙丸可缓解AA大鼠疼痛，其作用机制与调节AA大鼠垂体ACTH、β-EP的表达有关。
[Abstract]:Objective to investigate the effect of musk Oolong pills on the morphology of pituitary tissue and the expression of adrenocorticotropic hormone (ACTH) and beta endorphin (beta -endorphin, beta -EP) in the model of Adjuvant Arthritis (AA) model in rats, and to explore the town of musk Oolong pills in the treatment of rheumatoid arthritis (Rheumatoid arthritis, RA). The mechanism of pain action provides experimental and theoretical basis for its clinical application.
Methods after feeding 60 male Wistar rats to 7d, 12 rats were randomly selected as the normal group. Except for the normal group, the other 48 rats were anesthetized with ether, and the complete Freund adjuvant (Freund 's complete adjuvant, FCA) was injected into the right posterior foot. The model of AA rats was established, and the model was randomly selected for fifteenth days according to the arthritis index (arthritis index, AI). Divided into model group, musk Oolong pill low dose group, musk Oolong pill medium dose group, musk Oolong pill high dose group, 12 in each group, respectively give musk Oolong pill low, middle, high dose group of musk Oolong pill 0.3g. Kg-1,0.6g. Kg-1,1.2g. Kg-1 (physiological saline suspension is equivalent to 3,6,12 times of adult drug dosage, respectively), normal group and model group The normal saline was given 1 times a day for 2 weeks. After fifteenth days, all rats were killed and the pituitary was opened. The pituitary tissue morphology was observed by hematoxylin Yi Hong (HE) staining, and the changes of pituitary tissue morphology were observed under the light microscope. The changes of ACTH and beta -EP expression were observed by immunohistochemical method. Positive cell expression rate.
Results the effect of 1 on the general condition of rats: normal diet, bright fur and good mental state. After injection of FCA, the other rats were injected with foot swelling and left hind feet and front foot red and swollen, activities decreased, drinking water decreased, hair sparsely, luster, and difficult walking. The rats in the treatment group of musk Oolong pill were compared with model group during the administration. The general situation was better, the reaction was sensitive, the activity increased, the swelling of the ankle and the palmar palmar gradually lightened, and the lustrous.2 had a significant effect on the AI in rats: the AI of the rats in the model group was significantly higher than that in the normal group (P < 0.05). After the treatment, the musk Oolong pills were lower in the middle and higher groups (P < 0.05), especially in the high dose of musk Oolong pills. The effect of.3 on the morphology of pituitary tissue in rats was observed. Under the light microscope, HE staining was observed in each group. Compared with the normal group, the other groups of pituitary tissue did not significantly change the effect of.4 on the ACTH expression of pituitary gland: immunohistochemical method was used to observe the changes of ACTH expression in the pituitary tissue of each group, and the results of each group were ACTH. The positive rate of the cells in the normal group was 40.23 + 3.43, the model group was 56.82 + 3.97, the low dose group of musk Oolong pill was 53.14 + 4.16, the middle dose group of musk Oolong pill was 50.14 + 3.39, the high dose group of musk Oolong pill was 46.87 + 3.31., and the positive expression of the other 4 groups of ACTH cells increased obviously, and the difference was statistically significant (P < 0.01). Musk The positive expression of ACTH in the low dose group and the model group had no significant difference (P > 0.05). The positive expression of ACTH cells in the middle dose group of musk Oolong pill and the high dose group of musk Oolong pill was significantly lower than that of the model group. The difference was statistically significant (P < 0.05 or P < 0.01). The expression of the pituitary ACTH positive cells in the middle and high three groups of musk Oolong pills was low. In the high dose group of musk Oolong pill, the expression of ACTH expression was the most obvious effect of.5 on the expression of pituitary beta -EP: the expression of beta -EP in pituitary tissue was observed by immunohistochemistry. The positive rate of beta -EP cells in each group was 30.84 + 2.19 in the normal group, 39.72 in the model group, and the musk Oolong pill in the model group. The low dose group was 43.55 + 3.59, the middle dose group of musk Oolong pill was 46.69 + 3.62, the high dose group of musk Oolong pill was 49.47 + 3.17. compared to the normal group. The positive expression of beta -EP cells in the model group and the musk Oolong pill increased obviously, and the difference was statistically significant (P < 0.01). The positive expression of the musk Oolong pill was low, middle and high dose group. It was significantly higher than the model group, and the difference was statistically significant (P < 0.01). The effect of musk Oolong pill low, middle and high three groups on the expression of pituitary beta -EP positive cells was dose-dependent, and the expression of beta -EP in high dose group of musk Oolong pill was most obvious. Conclusion musk ulong pill can relieve pain in AA rats, its mechanism and regulation of AA are large. The expression of ACTH, beta -EP in the rat pituitary is related.
【学位授予单位】：河北联合大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R285.5

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