丹贝益肺方对肺纤维化大鼠肺组织中MEK、ERK及其磷酸化水平的影响

发布时间：2018-03-13 16:05

本文选题：特发性肺纤维化　切入点：丹贝益肺方　出处：《黑龙江省中医药科学院》2016年硕士论文　论文类型：学位论文

【摘要】：目的:本课题采用实验的方法通过博来霉素诱导致大鼠肺纤维化,观察经验方丹贝益肺方对该模型中肺纤维化大鼠的一般状态及肺组织形态的影响和对肺组织中MEK、ERK表达水平及其磷酸化水平的影响,证明丹贝益肺方在治疗疾病肺纤维化中对于一般临床表现的有效性及对肺组织中指标的影响。方法:选取SPF级Wistar大鼠90只,性别标准:雌雄各半,饲养要求:按照SPF级的标准进行适应性饲养1周,然后采用随机的方法将大鼠分为五组,分别为:正常空白对照组、模型对照组、泼尼松组、丹贝益肺方低剂量组、丹贝益肺方高剂量组,每组18只。按照完全随机的方法,将实验所用的大鼠依照文献中博莱霉素诱导造模的方法进行造模。大鼠造模成功后分别按照生理盐水,丹贝益肺水煎液0.3g/ml,丹贝益肺水煎液1.2g/ml,泼尼松溶液0.5mg/ml的标准进行灌胃,在造模成功后分别按不同的时间周期在第7、14、28天每次随机选取六只大鼠,先观察大鼠的一般状态的变化,然后处死大鼠,处死后迅速进行肺组织取材,先用肉眼观察每组大鼠的肺组织的外观的变化;再对肺组织处理行Masson染色的方法,然后在染色镜下观察肺的外观变化,并观察肺纤维化的程度,最后将肺组织制备成片,用SABC免疫组化法测定肺组织中MEK1/2、ERK1/2的水平,行Western blot法测定MEK、ERK的磷酸化水平。结果:丹贝益肺方高低剂量组及泼尼松组都能改善博莱霉素诱导肺纤维化大鼠的活动表现,用药组的大鼠一般情况均有所改善,程度不同,实验证实丹贝益肺方能够降低其肺组织中MEK1/2、ERK1/2及其磷酸化水平,染色镜下观察大鼠肺组织纤维化程度有所减轻。研究发现,丹贝益肺方可显著降低博莱霉素致大鼠的肺组织中MEK1/2、ERK1/2水平(P0.01),与强的松组作用相似(P0.05),丹贝益肺方高低剂量组之间的比较未见明显差异(P0.05)。丹贝益肺方可显著降低博莱霉素致肺纤维化大鼠的肺组织p-MEK1/2、p-ERK1/2水平(P0.01)。通过实验过程及所得结果表明,丹贝益肺方可以改善博莱霉素致大鼠肺纤维化的程度,减轻肺纤维化的临床症状,延缓肺纤维化进程;丹贝益肺方可以降低博莱霉素致肺纤维化大鼠的肺组织中MEK1/2、ERK1/2及其磷酸化水平。结论:1.经验方丹贝益肺方可降低肺纤维化大鼠的肺组织中MEK1/2、ERK1/2的水平以及其磷酸化水平的表达。2.丹贝益肺方能减缓肺纤维化进程的进展。
[Abstract]:Objective: to induce pulmonary fibrosis in rats by bleomycin. To observe the effects of Fang Dan Bei Yifei Fang on the general state of pulmonary fibrosis rats and the morphology of lung tissue and the expression level and phosphorylation level of MEKO ERK in lung tissue. To prove the effectiveness of Danbei Yifei recipe in the treatment of pulmonary fibrosis and its influence on the indexes of lung tissue. Methods: 90 Wistar rats of SPF grade were selected. Feeding requirements: according to the standard of SPF grade, the rats were fed adaptively for one week. Then the rats were randomly divided into five groups: normal blank control group, model control group, prednisone group, and low dose group. According to the totally random method, the rats used in the experiment were modeled according to the method of bleomycin induction in the literature. After the successful establishment of the model, the rats were made in accordance with normal saline, respectively. Danbei Yifei decoction 0.3 g / ml, Danbei Yifei decoction 1.2 g / ml, prednisone 0.5 mg / ml were administered by stomach. Six rats were randomly selected according to different time cycle at day 7 14: 28 to observe the changes of the general state of the rats. Then the rats were killed. After being killed, the lung tissues were collected quickly, and the appearance of each group of rats was observed with the naked eye. Then the method of Masson staining was used to treat the lung tissue, and then the appearance of the lung was observed under the staining microscope. The degree of pulmonary fibrosis was observed. At last, the lung tissue was prepared into pieces, and the level of MEK1 / 2 ERK1 / 2 in lung tissue was measured by SABC immunohistochemical method. Results: the activity of bleomycin induced pulmonary fibrosis was improved in the high and low dose group and prednisone group. The experiment proved that Danbei Yifei Fang could reduce MEK1 / 2 ERK1 / 2 and its phosphorylation level in lung tissue, and observe the degree of pulmonary fibrosis in rats by staining microscope. Danbei Yifei decoction significantly reduced the level of MEK1 / 2 ERK1 / 2 in the lung tissue of rats induced by bleomycin, which was similar to that in prednisone group (P 0.05). There was no significant difference between the high and low dose groups of Danbei Yifei recipe. The levels of p-MEK1 / 2 and p-ERK1 / 2 in pulmonary tissue of rats with pulmonary fibrosis induced by mycin were P0.01. the experimental process and the results showed that. Danbei Yifei prescription can improve the degree of pulmonary fibrosis induced by bleomycin in rats, alleviate the clinical symptoms of pulmonary fibrosis and delay the progress of pulmonary fibrosis. Danbei Yifei recipe can reduce MEK1 / 2 ERK1 / 2 and its phosphorylation level in lung tissue of bleomycin-induced pulmonary fibrosis rats. Conclusion: 1. Experience Fang bai Yifei decoction can reduce the level of MEK1 / 2ERK1 / 2 and its phosphorylated water in lung tissue of rats with pulmonary fibrosis. Ping expression. 2. Danbei Yifei Fang can slow the progression of pulmonary fibrosis.
【学位授予单位】：黑龙江省中医药科学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R259

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