本文選題：特發(fā)性肺間質(zhì)纖維化 + 博萊霉素　； 參考：《湖北民族學院》2014年碩士論文

【摘要】：目的： 本實驗采用腹腔注射麻醉并氣管注入博來霉素建立大鼠肺間質(zhì)纖維化模型,從形態(tài)組織及細胞學的角度觀察并分析益氣養(yǎng)陰方對博萊霉素致大鼠肺間質(zhì)纖維化的影響,研究益氣養(yǎng)陰方對肺間質(zhì)纖維化的治療作用,為其進一步的臨床運用及民族藥物新制劑的開發(fā)奠定基礎(chǔ),為臨床治療肺間質(zhì)纖維化提供理論依據(jù)。 方法： 清潔級健康雄性SD大鼠60只,體重為240±10g,隨機分為5組：正常組、模型組、中藥組、激素組、中藥+激素組。正常組氣管內(nèi)注入生理鹽水,其余4組分別注入相應(yīng)劑量博萊霉素建立肺間質(zhì)纖維化大鼠模型。造模后第28天開始灌胃給藥,每天一次。中藥組、激素組、中藥+激素組分別灌入約2ml相應(yīng)濃度的藥物溶液,正常組與模型組灌入2ml生理鹽水,連續(xù)灌胃28天。各組動物于灌胃后第14天、28天分兩批處死。處死后分離肺組織,肉眼觀察肺組織外觀形態(tài)；分別取左肺下葉同一部位行病理切片,作HE染色和Masson染色以觀察形態(tài)學改變；同時取右肺底同一部位用4%多聚甲醛固定后,用原位雜交檢測試劑盒檢測肺組織中TNF-α、TGF-β1、IFN-γ的表達,觀察益氣養(yǎng)陰方對肺間質(zhì)纖維化的治療效果,并行統(tǒng)計學分析。 結(jié)果： 1一般狀態(tài)觀察：正常組飲食如常,好動活潑,反應(yīng)敏捷,皮毛有光澤,體重逐漸增加,二便正常,第28天時無死亡現(xiàn)象；模型組食欲不振,精神萎靡,反應(yīng)遲鈍,蜷縮少動,多有皮毛倒豎現(xiàn)象,身體較其它組消瘦,且均有不同程度的咳嗽,二便減少,28天時死亡4只；中藥組飲食可,反應(yīng)靈敏,體重無明顯減輕,二便正常,28天時無死亡現(xiàn)象；激素組及中藥+激素組飲食、皮毛尚可,體重有所減輕,28天時各有2只死亡。給藥第0d即造模后第28天,模型組、中藥組、激素組及中藥+激素組與正常組相比,體重明顯降低,且有明顯差異(P0.05)；給藥后第28d中藥組與激素組體重增長高于模型組,且有統(tǒng)計學意義(P0.05)。 2肺指數(shù)觀察：給藥28d后激素組、中藥組及中藥+激素組肺指數(shù)相比于模型組均下降,且具有統(tǒng)計學意義(P0.05)；模型組大鼠與正常組相比肺指數(shù)升高,且有統(tǒng)計學意義(P0.05)。 3肺組織中TNF-αmRNA、TGF-β1mRNA、IFN-γmRNA的表達 3.1模型組與正常組相比,TNF-αmRNA陽性細胞率升高,且具有統(tǒng)計學意義（P0.05）；激素組、中藥組、中藥+激素組與模型組相比,TNF-αmRNA陽性細胞率降低,且具有統(tǒng)計學意義（P0.05）；中藥組與激素組、中藥+激素組相比,TNF-αmRNA陽性細胞率低,且具有統(tǒng)計學意義（P0.05）。 3.2正常組、激素組、中藥組以及中藥+激素組與模型組相比,TGF-β1mRNA陽性細胞率降低,且具有統(tǒng)計學意義（P0.05）；中藥組與激素組相比較,TGF-β1mRNA陽性細胞率略低,且具有統(tǒng)計學意義（P0.05）；中藥組與中藥+激素組比較,無統(tǒng)計學意義（P0.05）。 3.3正常組、激素組、中藥組、中藥+激素組和模型組相比,IFN-γmRNA陽性細胞率升高,且具有統(tǒng)計學意義（P0.05）；中藥組與激素組、中藥+激素組相比,IFN-γmRNA陽性細胞率略高,且具有統(tǒng)計學意義（P0.05）。 結(jié)論： 1大鼠氣管內(nèi)注射博來霉素可建立肺間質(zhì)纖維化大鼠模型,本實驗肺組織病理學觀察及肺組織中相關(guān)細胞因子檢測驗證了大鼠模型復制成功。 2益氣養(yǎng)陰組方作為附院防治IPF的經(jīng)驗方,臨床已應(yīng)用多年,本實驗通過復制IPF大鼠模型,檢測相關(guān)細胞因子,了解其防治IPF的作用機理,發(fā)現(xiàn)其可能通過降低肺組織中TNF-α、TGF-β1的含量和升高肺組織內(nèi)IFN-γ的含量,最終達到延緩肺纖維化的作用。 3中醫(yī)藥防治IPF已經(jīng)展示出了良好的發(fā)展前景,有待于大家進一步研究。
[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.
【學位授予單位】：湖北民族學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R259

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本文編號：2052204