益气养阴法对肺间质纤维化大鼠TNF-α、TGF-β1、IFN-γ作用的影响研究

发布时间：2018-06-22 08:19

本文选题：特发性肺间质纤维化 + 博莱霉素　；参考：《湖北民族学院》2014年硕士论文

【摘要】：目的：本实验采用腹腔注射麻醉并气管注入博来霉素建立大鼠肺间质纤维化模型,从形态组织及细胞学的角度观察并分析益气养阴方对博莱霉素致大鼠肺间质纤维化的影响,研究益气养阴方对肺间质纤维化的治疗作用,为其进一步的临床运用及民族药物新制剂的开发奠定基础,为临床治疗肺间质纤维化提供理论依据。方法：清洁级健康雄性SD大鼠60只,体重为240±10g,随机分为5组：正常组、模型组、中药组、激素组、中药+激素组。正常组气管内注入生理盐水,其余4组分别注入相应剂量博莱霉素建立肺间质纤维化大鼠模型。造模后第28天开始灌胃给药,每天一次。中药组、激素组、中药+激素组分别灌入约2ml相应浓度的药物溶液,正常组与模型组灌入2ml生理盐水,连续灌胃28天。各组动物于灌胃后第14天、28天分两批处死。处死后分离肺组织,肉眼观察肺组织外观形态；分别取左肺下叶同一部位行病理切片,作HE染色和Masson染色以观察形态学改变；同时取右肺底同一部位用4%多聚甲醛固定后,用原位杂交检测试剂盒检测肺组织中TNF-α、TGF-β1、IFN-γ的表达,观察益气养阴方对肺间质纤维化的治疗效果,并行统计学分析。结果： 1一般状态观察：正常组饮食如常,好动活泼,反应敏捷,皮毛有光泽,体重逐渐增加,二便正常,第28天时无死亡现象；模型组食欲不振,精神萎靡,反应迟钝,蜷缩少动,多有皮毛倒竖现象,身体较其它组消瘦,且均有不同程度的咳嗽,二便减少,28天时死亡4只；中药组饮食可,反应灵敏,体重无明显减轻,二便正常,28天时无死亡现象；激素组及中药+激素组饮食、皮毛尚可,体重有所减轻,28天时各有2只死亡。给药第0d即造模后第28天,模型组、中药组、激素组及中药+激素组与正常组相比,体重明显降低,且有明显差异(P0.05)；给药后第28d中药组与激素组体重增长高于模型组,且有统计学意义(P0.05)。 2肺指数观察：给药28d后激素组、中药组及中药+激素组肺指数相比于模型组均下降,且具有统计学意义(P0.05)；模型组大鼠与正常组相比肺指数升高,且有统计学意义(P0.05)。 3肺组织中TNF-αmRNA、TGF-β1mRNA、IFN-γmRNA的表达 3.1模型组与正常组相比,TNF-αmRNA阳性细胞率升高,且具有统计学意义（P0.05）；激素组、中药组、中药+激素组与模型组相比,TNF-αmRNA阳性细胞率降低,且具有统计学意义（P0.05）；中药组与激素组、中药+激素组相比,TNF-αmRNA阳性细胞率低,且具有统计学意义（P0.05）。 3.2正常组、激素组、中药组以及中药+激素组与模型组相比,TGF-β1mRNA阳性细胞率降低,且具有统计学意义（P0.05）；中药组与激素组相比较,TGF-β1mRNA阳性细胞率略低,且具有统计学意义（P0.05）；中药组与中药+激素组比较,无统计学意义（P0.05）。 3.3正常组、激素组、中药组、中药+激素组和模型组相比,IFN-γmRNA阳性细胞率升高,且具有统计学意义（P0.05）；中药组与激素组、中药+激素组相比,IFN-γmRNA阳性细胞率略高,且具有统计学意义（P0.05）。结论： 1大鼠气管内注射博来霉素可建立肺间质纤维化大鼠模型,本实验肺组织病理学观察及肺组织中相关细胞因子检测验证了大鼠模型复制成功。 2益气养阴组方作为附院防治IPF的经验方,临床已应用多年,本实验通过复制IPF大鼠模型,检测相关细胞因子,了解其防治IPF的作用机理,发现其可能通过降低肺组织中TNF-α、TGF-β1的含量和升高肺组织内IFN-γ的含量,最终达到延缓肺纤维化的作用。 3中医药防治IPF已经展示出了良好的发展前景,有待于大家进一步研究。
[Abstract]:Objective:
In this experiment, a rat model of pulmonary fibrosis was established by intraperitoneal injection and intratracheal injection of bleomycin. The effects of Yiqi Yangyin prescription on pulmonary fibrosis in rats induced by bleomycin were observed and analyzed from the angle of morphology and cytology, and the therapeutic effect of Yiqi Yangyin prescription on pulmonary fibrosis was further studied. It will lay a foundation for the development of new preparations of ethnic medicines and provide a theoretical basis for clinical treatment of pulmonary interstitial fibrosis.
Method:
60 healthy male SD rats, with a weight of 240 + 10g, were randomly divided into 5 groups: normal group, model group, Chinese medicine group, hormone group, Chinese medicine + hormone group. Normal saline was injected into the normal group, and the other 4 groups were injected with bleomycin to establish pulmonary fibrosis rat model respectively. After twenty-eighth days after the model, the rats were given the medicine, once a day. The Chinese medicine group, the hormone group, the Chinese medicine + hormone group were injected with the corresponding concentration of 2ml. The normal group and the model group were perfused with 2ml saline for 28 days. The animals were sacrificed at fourteenth days after gavage and 28 days in two batches. The lung tissue was separated and the appearance of the lung tissue was observed by the naked eye; the left lower lobe was taken from the same site in the same part of the lung. The morphological changes were observed by HE staining and Masson staining. At the same time, the same part of the right lung was fixed with 4% polyformaldehyde. In situ hybridization detection kit was used to detect the expression of TNF- alpha, TGF- beta 1, IFN- gamma in lung tissue, and the therapeutic effect of Yiqi Yangyin prescription on pulmonary fibrosis was observed, and the statistical analysis was carried out.
Result:
1 general state observation: normal diet, active and active, active and active, quick reaction, luster, weight gradually increasing, two normal, twenty-eighth days without death; model group loss of appetite, mental malaise, slow reaction, curled up less movement, more skin and vertical phenomenon, body thinner than the other groups, and have different degrees of coughing and two stool There were 4 deaths at 28 days. The diet of the Chinese medicine group was sensitive, the weight was not obviously reduced, the two was normal, and there was no death at 28 days. The diet of the hormone group and the Chinese medicine + hormone group, the fur and the weight were reduced, and 2 died in 28 days. The drug group, 0d, the model group, the Chinese medicine group, the hormone group and the traditional Chinese medicine + hormone group and the normal group. Compared with the control group, the body weight was significantly decreased, and there was a significant difference (P0.05). The weight gain of the 28d group and the hormone group was higher than that of the model group after the administration, and the difference was statistically significant (P0.05).
2 lung index observation: after the administration of 28d, the lung index of the Chinese medicine group and the Chinese medicine + hormone group decreased compared with the model group, and had statistical significance (P0.05). The lung index of the model group was higher than that of the normal group, and had statistical significance (P0.05).
3 expression of TNF- alpha mRNA, TGF- beta 1mRNA and IFN- gamma mRNA in lung tissue
3.1 compared with the normal group, the rate of TNF- alpha mRNA positive cells increased and had statistical significance (P0.05). Compared with the model group, the rate of TNF- alpha mRNA positive cells in the hormone group, Chinese medicine group, Chinese medicine + hormone group decreased and had statistical significance (P0.05). The rate of TNF- alpha mRNA positive cells in the Chinese medicine group and the irritable group, the Chinese traditional medicine + hormone group, and the TNF- alpha mRNA positive cells were low, and had a low rate of TNF- alpha mRNA positive cells. There was a statistical significance (P0.05).
3.2 the normal group, the hormone group, the traditional Chinese medicine group and the Chinese medicine + hormone group compared with the model group, the rate of TGF- beta 1mRNA positive cells decreased and had statistical significance (P0.05). Compared with the hormone group, the rate of TGF- beta 1mRNA positive cells was slightly lower and had statistical significance (P0.05), and there was no statistical significance (P0.05) compared with the Chinese traditional medicine group and the Chinese medicine + hormone group.
3.3 the normal group, the hormone group, the traditional Chinese medicine group, the Chinese medicine + hormone group and the model group, compared with the model group, the rate of IFN- gamma mRNA positive cells increased and had statistical significance (P0.05). Compared with the hormone group, Chinese medicine + hormone group, the rate of IFN- gamma mRNA positive cells was slightly higher and had statistical significance (P0.05).
Conclusion:
The 1 rat models of intratracheal intratracheal injection of bleomycin can establish a rat model of pulmonary fibrosis. In this experiment, the pathological observation of lung tissue and the detection of related cytokines in the lung tissue proved that the rat model was replicated successfully.
2 Yiqi nourishing Yin group as the experience of IPF in the hospital has been used for many years. In this experiment, the IPF rat model was used to detect the related cytokines and understand the mechanism of its prevention and control of IPF. It was found that it could reduce the content of TNF- alpha, TGF- beta 1 and increase the content of IFN- gamma in lung tissue, and finally reached the delay of lung fiber. The role of chemistry.
3 the prevention and treatment of IPF by traditional Chinese medicine has shown good prospects for further study.
【学位授予单位】：湖北民族学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R259

【参考文献】