本文選題：肺間質(zhì)纖維化 + 博萊霉素��； 參考：《河北醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的:肺間質(zhì)纖維化(pumlonary interstitial fibrosis)是一種慢性漸進(jìn)性疾病,以炎癥反應(yīng)、成纖維細(xì)胞增殖和細(xì)胞外基質(zhì)蛋白沉積為特點(diǎn),誘因眾多。而特發(fā)性肺間質(zhì)纖維化(idiopathic pulmonary fibrosis,IPF)被定義為一種特殊形式的慢性纖維變性的間質(zhì)性肺炎,組織病理學(xué)表現(xiàn)為尋常性間質(zhì)肺炎�，F(xiàn)已證實(shí)它是一種限制性通氣功能障礙和彌散功能障礙疾病,患者最終死于呼吸衰竭。盡管IPF預(yù)后不良,但發(fā)病機(jī)制仍為未知,并且尚無(wú)有效的治療措施。目前的治療方法通常包括皮質(zhì)類固醇激素或聯(lián)合使用一種細(xì)胞毒藥物。但是免疫抑制療法的作用有限,并且副作用不容忽視。因此,迫切需要一種替代治療方案。人肺成纖維細(xì)胞的體外實(shí)驗(yàn)研究表明,沙利度胺能減少IL-6、TGF-β_1、VEGF、Ang-1的產(chǎn)生及膠原的合成,并能抑制依賴IL-6的成纖維細(xì)胞增殖和依賴TGF-β_1的成纖維細(xì)胞轉(zhuǎn)分化,這些可能是沙利度胺預(yù)防肺纖維化的潛在作用機(jī)制。 本實(shí)驗(yàn)旨在研究沙利度胺對(duì)博來(lái)霉素致大鼠纖維化的干預(yù)作用。通過(guò)測(cè)量肺組織內(nèi)羥脯氨酸(HYP)含量,以及肺組織和血清中TNF-α、TGF-β_1的表達(dá)水平探討可能的作用機(jī)制。 方法:選取清潔級(jí)健康、雄性SD大鼠(河北醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物中心提供)90只,體重220±10g,適應(yīng)性飼養(yǎng)一周后,隨機(jī)分為:(1)對(duì)照組(A組):30只,氣管內(nèi)注入0.1ml/100g生理鹽水,于假造模后第一日用生理鹽水1ml/100g灌胃,1次/d,分別于7、14、28天各處死10只。(2)模型組(B組):30只,氣管內(nèi)一次性注入博來(lái)霉素A_5(5mg/kg),于造模后第一日用生理鹽水1ml/100g灌胃,1次/d,分別于7、14、28天各處死9只、10只、9只。(3)沙利度胺組(C組):30只,氣管內(nèi)一次性注入博來(lái)霉素A_5(5mg/kg),于造模后第一日用沙利度胺溶液(沙利度胺與生理鹽水配成1%溶液)100mg/kg灌胃,1次/d,分別于7、14、28天各處死10只。各組剪斷雙側(cè)股動(dòng)脈取血并放血處死,用酶聯(lián)免疫吸附試驗(yàn)(ELISA)法測(cè)定血清中TNF-α、TGF-β_1的含量。取右肺下葉固定于4%多聚甲醛溶液中,用免疫組化法檢測(cè)TNF-α、TGF-β_1,并用圖像分析系統(tǒng)半定量測(cè)定TNF-α和TGF-β_1的陽(yáng)性面積百分比。通過(guò)HE染色、Masson膠原染色來(lái)評(píng)價(jià)肺泡炎和肺纖維化的程度,并按照Szapiel法分為4級(jí)。右肺上葉制成勻漿測(cè)定肺組織中羥脯氨酸濃度。采用SPSS13.0統(tǒng)計(jì)軟件處理數(shù)據(jù)。 結(jié)果: 1肺組織病理分析結(jié)果:在同一時(shí)間點(diǎn),B組肺泡炎程度顯著高于A組(P0.01),B組肺纖維化程度亦顯著高于A組,7天時(shí)(P0.05),14天、28天時(shí)(P0.01),表明動(dòng)物模型復(fù)制成功。C組肺泡炎及纖維化程度較模型組減輕,7天:C組肺泡炎程度較B組顯著減輕(P0.01);14天:C組肺泡炎及纖維化程度較B組顯著減輕(P0.01);28天:C組纖維化程度較B組顯著減輕(P0.01)。 2肺組織羥脯氨酸(HYP)含量變化:B組大鼠肺組織HYP含量隨造模時(shí)間延長(zhǎng)逐漸升高,至28天達(dá)高峰,7天時(shí)與A組相比差異有統(tǒng)計(jì)學(xué)意義(P0.01),與A組相比各時(shí)間點(diǎn)HYP含量的差異均具有統(tǒng)計(jì)學(xué)意義(P0.01)。C組大鼠HYP含量呈低水平升高趨勢(shì),造模后7、14、28天HYP含量均顯著低于B組(P0.01)。 3肺組織TNF-α,TGF-β_1含量變化:B組大鼠肺組織中TNF-α、TGF-β_1表達(dá)量于7、14、28天均高于A組、C組,各時(shí)間點(diǎn)比較均有統(tǒng)計(jì)學(xué)意義(P0.01)。 4血清中TNF-α,TGF-β_1含量變化:B組大鼠血清中TNF-α的表達(dá)量除28天外,其余各時(shí)間點(diǎn)均明顯高于A和C組,差異具有統(tǒng)計(jì)學(xué)意義(P0.01);B組大鼠血清中TGF-β_1表達(dá)量于7、14、28天均高于A組和C組,各時(shí)間點(diǎn)比較均有統(tǒng)計(jì)學(xué)意義(P0.01)。 結(jié)論: 1沙利度胺在博萊霉素誘導(dǎo)的大鼠肺纖維化模型中,能夠減少肺間質(zhì)的膠原沉積,對(duì)肺間質(zhì)纖維化具有一定的治療作用。 2沙利度胺對(duì)TNF-α和TGF-β_1的產(chǎn)生具有抑制作用,進(jìn)而減輕早期的炎癥反應(yīng),抑制晚期的纖維化改變,這可能是沙利度胺對(duì)肺纖維化防治作用的潛在機(jī)制。
[Abstract]:Objective: pulmonary interstitial fibrosis (pumlonary interstitial fibrosis) is a chronic progressive disease characterized by inflammatory response, fibroblast proliferation and extracellular matrix protein deposition. And idiopathic pulmonary fibrosis (idiopathic pulmonary fibrosis, IPF) is defined as a special form of chronic fibrous degeneration. Interstitial pneumonia, histopathological manifestation of common interstitial pneumonia. It has been proved to be a restrictive ventilation dysfunction and diffuse dysfunction disease, and the patient eventually died of respiratory failure. Although the prognosis of IPF is poor, the pathogenesis is still unknown, and there is no effective treatment. The current treatment methods usually include cortex. Steroid hormones or combined use of a cytotoxic drug. But immunosuppressive therapy is limited and side effects can not be ignored. Therefore, an alternative therapy is urgently needed. In vitro experimental study on human lung fibroblasts showed that thalidomide could reduce the production of IL-6, TGF- beta _1, VEGF, Ang-1, and collagen synthesis and inhibit the dependence of human lung fibroblasts. The proliferation and the fibroblast transdifferentiation of TGF- beta _1 may be the potential mechanism of thalidomide in preventing pulmonary fibrosis in Lai IL-6.
The aim of this study was to investigate the effect of thalidomide on bleomycin induced fibrosis in rats. The possible mechanism of the expression of hydroxyproline (HYP) in lung tissue and the expression level of TNF- alpha and TGF- beta _1 in lung tissue and serum was investigated.
Methods: 90 male SD rats (Hebei Medical University experimental animal center), weighing 220 + 10g, were randomly divided into two groups: (1) the control group (group A): 30 rats were injected with 0.1ml/100g saline in the trachea. The first day after the artificial model was given to the stomach with physiological saline 1ml/100g and 1 times /d, respectively, to death 1 days, respectively, were killed 1 days respectively, and respectively to the 7,14,28 days to death 1 respectively. 0 (2) the model group (group B): 30 rats were injected with bleomycin A_5 (5mg/kg) in the trachea at one time. After the first day, the normal saline 1ml/100g was administered to the stomach, and 1 times /d, respectively, on 7,14,28 days, respectively, and 9. (3) the thalidomide group (group C): 30, and the bleomycin A_5 (5mg/kg) was injected into the trachea for the first time, and the first day was dissolved in thalidomide after the model. The solution (thalidomide and physiological saline mixed with 1% solution) was gavage with 100mg/kg, 1 times /d, and 10 rats were killed on 7,14,28 days respectively. Each group was cut off the bilateral femoral arteries to take blood and put to death. The content of TNF- alpha and TGF- beta _1 in serum was determined by enzyme linked immunosorbent assay (ELISA). The right lower lobe was fixed in 4% polycondensation Formaldehyde Solution and examined by immunohistochemical method. TNF- alpha, TGF- beta _1 were measured and the percentage of positive area of TNF- A and TGF- beta _1 was semi quantified by image analysis system. The degree of pulmonary alveolitis and pulmonary fibrosis was evaluated by HE staining and Masson collagen staining, and divided into 4 grades according to Szapiel method. The concentration of hydroxyproline in lung tissue was measured by the upper lobe of right lung, and SPSS13.0 statistics software was used. Data.
Result:
1 pathological analysis of lung tissue: at the same time point, the degree of alveolitis in group B was significantly higher than that in group A (P0.01), and the degree of pulmonary fibrosis in group B was also significantly higher than that in group A, 7 days (P0.05), 14 days, and 28 days (P0.01), indicating that animal model replicating of pulmonary alveolitis and fibrosis process was less than that in model group, 7 days: the degree of pulmonary alveolitis in C group was significantly less than that in B group (P0.0) 1); 14 days: alveolar inflammation and fibrosis in group C were significantly lower than those in group B (P0.01); on the 28 day: fibrosis in group C was significantly lower than that in group B (P0.01).
2 changes in the content of hydroxyproline (HYP) in lung tissue: the HYP content in lung tissue of rats in group B increased gradually with the duration of mould making, to the peak of 28 in Tianda. The difference between the 7 days and the A group was statistically significant (P0.01). The difference of HYP content at each time point compared with the A group had statistical significance (P0.01) the HYP content of the.C group was low level, and the model was created. The content of HYP in the post 7,14,28 days was significantly lower than that in the B group (P0.01).
3 the changes in the content of TNF- alpha and TGF- beta _1 in lung tissue: the expression of TNF- alpha and TGF- beta _1 in the lung tissue of the rats in group B was higher than that in the A group, and the C group was statistically significant (P0.01) at all time points.
4 the changes of the content of TNF- A and TGF- beta _1 in serum: the expression of TNF- alpha in the serum of B rats was significantly higher than that of the A and C groups, and the difference was statistically significant (P0.01). The _1 expression of TGF- beta in the serum of the group B rats was higher than that in the 7,14,28 day group and the group, and the time points were statistically significant.
Conclusion:
1 thalidomide in bleomycin induced rat pulmonary fibrosis model can reduce the collagen deposition in the interstitial lung, and have a certain therapeutic effect on pulmonary fibrosis.
2 thalidomide inhibits the production of TNF- A and TGF- beta _1, thus alleviates the early inflammatory response and inhibits late fibrosis, which may be a potential mechanism for the prevention and treatment of pulmonary fibrosis by thalidomide.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R965

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 楊云;張王剛;陳銀霞;曹星梅;何愛麗;楊惠云;田瑋;;沙利度胺對(duì)人外周血單個(gè)核細(xì)胞免疫正調(diào)控作用[J];中國(guó)實(shí)驗(yàn)血液學(xué)雜志;2006年06期

2 宋明臣,何冰,邱忠民,李海潮;肺纖維化大鼠肺泡Ⅱ型細(xì)胞中細(xì)胞因子的表達(dá)[J];中華結(jié)核和呼吸雜志;1998年04期

3 陳巍,李振華,侯顯明,于潤(rùn)江;間質(zhì)性肺疾病支氣管肺泡灌洗液中性粒細(xì)胞趨化因子和腫瘤壞死因子水平及其臨床意義[J];中華結(jié)核和呼吸雜志;1998年05期

4 曹彬,郭子健,許文兵,朱元玨;細(xì)胞因子在特發(fā)性肺間質(zhì)纖維化血管生成中的作用[J];中華內(nèi)科雜志;1999年12期

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