博莱霉素致大鼠肺纤维化胶原表达的研究

目的：探讨博莱霉素致大鼠肺间质纤维化肺组织前胶原I和前胶原III mRNA及蛋白质变化的意义。方法：大鼠随机分为经气管内一次性灌注生理盐水为正常对照组,灌注搏莱霉素分为博莱霉素3天组,7天组,14天组,30天组,分子杂交和免疫组化方法测定肺组织的前胶原I,前胶原III的mRNA和蛋白质表达。结果：博莱霉素7天组前胶原I,前胶原III的mRNA高于正常对照组（P＜0．01）,14天组达到峰值水平,博莱霉素30天组的前胶原I mRNA仍高于正常对照组（P＜0．01）,而前胶原III的mRNA水平与正常对照组。结论：博莱霉素致肺纤维化时,前胶原基因转录水平的升高,引起的胶原合成加速是肺间质胶原积聚的主要原因。     

实验性肺纤维化中胶原基因表达的动态变化

采用博莱霉素复制大鼠肺纤维化模型，分别于第２周和第４周分两批处死大鼠，观察Ｉ、Ⅲ型胶原基因的动态变化。结果显示：在第２周时，博莱霉素组的组织病理学为典型的Ⅲ级肺泡炎表现，成纤维细胞增生。用Ｐｒｏα１（Ｉ）、Ｐｒｏα１（Ⅲ）肽莲的ｃＤＮＡ探针进行ＤＮＡ－ＲＮＡ斑点杂交显示，Ｐｒｏα１（Ｉ）ｍＲＮＡ表达水平在博莱霉素组、地塞米松治疗组均明显高于正常对照组（Ｐ〈０．０１）。在第４周时，博莱霉素组为典型的     

博莱霉素致大鼠外周血淋巴细胞DNA损伤的研究

为了探讨博莱霉素（BLM）的遗传损伤效应，我们应用单细胞微量凝胶电泳技术对抗癌药博莱霉素的DNA损伤作用进行了实验研究，现将结果报道如下。材料与方法一、动物染毒：80～100克体重雄性Fisher大鼠16只，随机分成两组，每组8只。实验组动物腹腔注射BLM6mg／kg，每天一次，每周用5天，连续6周。阴性对照组注射等量的0．9％NaCl。染毒结束后立即取静脉血3ml。二、淋巴细胞分离制备：在3ml全血中加入淋巴细胞分离液一离心制备得淋巴细胞是液，浓度调节为1X106／ml．台盼蓝法测定细胞存活率。三、单细胞微量凝胶电泳测试（MGE）程序：…     

氨基胍对博莱霉素诱发大鼠肺毒性的抑制作用

目的:观察氨基胍(AG)对博莱霉素A_5(BLMA_5)诱发大鼠肺毒性作用的影响。方法:BLM-A_5(5mg·kg~(-1))气管滴注诱发大鼠肺损伤和肺纤维化形成;在气管滴注BLM-A_5的同时及之后21d腹腔注射AG(20mg·kg~(-1)·d~(-1),ip)。化学比色法测定肺组织羟脯氨酸含量、肺泡巨噬细胞培养上清液中NO_2~-/NO_3~-含量、出肺血中NO_2~-/NO_3~-含量;HE染色组织切片上观察组织形态学变化。结果:(1)滴注BLM-A_5后d14至30,肺组织羟脯氨酸含量和出肺血中的MDA含量逐渐升高;注BLMA_5后d30,肺间质可见萎陷的和纤维化的肺泡,并出现大量的成纤维细胞。(2)滴注BLM-A_5后d7至14,出肺血NO_2~-/NO_3~-含量升高;肺泡灌流液中巨噬细胞培养上清液中NO_2~-/NO_3~-含量升高。(3)AG明显抑制出肺血MDA含量和肺羟脯氨酸含量的升高;AG还减轻肺间质组织形态学变化。(4)AG明显抑制出肺血NO_2~-/NO_3~-含量的升高。结论:AG对BLM-A_5诱发的大鼠肺损伤和纤维化有抑制作用,此作用与其抑制肺内NO大量生成有关。     

内源性过氧亚硝基阴离子在博莱霉素A5诱发大鼠肺损伤和肺纤维化中的作用

目的：观察内源性过氧亚硝基阴离子(ONOO^-)在博莱霉素A5诱发大鼠肺损伤和肺纤维化中的作用．方法：通过测定出肺血脂质过氧化物水平和观察肺组织学变化(包括用偏振光显微镜观察天狼猩红染色的I、Ⅲ型胶原的变化)来判断肺损伤和肺纤维化;用硝基酪氨酸的免疫组化判断过氧亚硝基阴离子的表达．结果：气管内给予博莱霉素A5第14天观察到(1)出肺血脂质过氧化物含量升高;肺泡壁增厚,肺间质巨噬细胞浸润,且其邻近有成纤维细胞及增多的I、Ⅲ型胶原．(2)肺泡上皮细胞和肺间质细胞内ONOO^-高表达．(3)诱导型一氧化氮合酶(iNOS)抑制剂氨基胍减轻上述变化．结论：内源性ONOO^-介导BLM-A5的肺毒性作用;氨基胍对肺损伤和肺纤维化的治疗部分是通过减少内源性ONOO^-的形成实现的．     

地塞米松地实验性肺纤维化的保护作用

以博莱霉素气管内给药复制大鼠肺纤维化模型，并给予地塞米松腹腔注射观察其抗肺纤维化的作用，于注药后第２周和第４周博莱霉素组的肺系数，ＨＰ含量均明显增６高，病理有典型的肺泡炎和肺纤维化表现。     

槲皮素对博莱霉素致鼠肺纤维化的防治作用

目的　观察SD大白鼠腹腔内注射槲皮素 ( 2 0 0mg·kg-1)对实验性博莱霉素致肺纤维化的防治作用。方法　通过博莱霉素 ( 5mg·kg-1)致鼠肺纤维化的动物模型 ,动态观察各实验组与肺纤维化形成相关的d 3、d 7、d 14、d 2 8肺组织匀浆内羟脯氨酸、脂质过氧化物 (LPO)含量 ,动态观察肺组织病理学变化及形态学定量测量肺组织中炎症组织数 /每高倍视野、肺泡间隔宽度、单位面积肺纤维化灶所占的分面积。结果　与对照组相比 ,槲皮素组能显著地降低博莱霉素致肺纤维化程度。结论　槲皮素对博莱霉素致肺纤维化有一定的防治作用     

表没食子儿茶素没食子酸酯对博莱霉素所致大鼠肺纤维化的干预作用及其机制研究

目的观察表没食子儿茶素没食子酸酯(EGCG)对实验性大鼠肺纤维化的干预作用及可能的作用机制。方法大鼠随机分为正常对照组、模型组、醋酸泼尼松组和EGCG大、中、小剂量组。通过气管内注入博莱霉素(BLM)复制大鼠肺纤维化模型,于造模后d 2各治疗组开始给药,给药后d7、14、28处死大鼠,取肺组织,观察形态学变化,并测定生化指标。结果EGCG能减少实验性肺纤维化大鼠肺组织中胶原沉积及降低肺系数(P<0.05,P<0.01),提高T-AOC、SOD水平(P<0.05,P<0.01),减轻肺部的病理损害。结论EGCG对BLM诱导产生的大鼠肺纤维化有一定的抑制作用。     

昆布提取物抑制大鼠肺纤维化的研究

目的研究昆布提取物(Okam)对博莱霉素(Bleomycin)所致大鼠肺间质纤维化的抑制作用。方法Wistar大鼠经气管内注入博莱霉素诱导肺纤维化,随后每日灌胃给予Okam 25、50 mg·kg~(-1)进行干预。通过免疫组化检测胶原Ⅲ、TNF—α和血管内皮生长因子(VEGF)蛋白在大鼠肺组织的表达。结果Okam 50mg·kg~(-1)可明显降低胶原Ⅲ、TNF-α和VEGF蛋白表达;Okam 25 mg·kg~(-1)可明显降低胶原Ⅲ、VEGF蛋白表达,但TNF-α表达与模型组比较无差异。结论昆布提取物灌胃给药能减轻博莱霉素诱导的大鼠肺纤维化的程度,有可能被开发成为治疗肺纤维化的海洋药物。     

黄芩总黄酮对博莱霉素致大鼠肺纤维化的干预作用

目的探讨黄芩总黄酮(total flavonoids of scutellariabaicalensis georgi,TFSB)对博莱霉素致大鼠肺纤维化的干预作用。方法气管内注入博莱霉素制备大鼠肺纤维化模型,给予TFSB灌服28 d,每天1次后,计算肺系数、测定肺组织和血清中肺纤维化部分相关生化指标,观察大鼠肺泡炎和肺纤维化程度,RT-PCR检测转化生长因子-β1(TGF-β1)、Smad2、Smad3 mRNA的表达水平,Western blot检测平滑肌肌动蛋白α(α-SMA)与过氧化物酶体增殖因子活化受体γ(PPAR-γ)的蛋白表达水平。结果与模型组相比,TFSB各剂量组大鼠肺组织羟脯氨酸(HYP)含量、血清与肺组织匀浆中丙二醛(MDA)含量降低(P<0.05,P<0.01),血清与肺组织匀浆中谷胱甘肽(GSH)含量、血清超氧化物歧化酶(SOD)活性、血清总抗氧化能力(T-AOC)升高(P<0.05,P<0.01),TFSB各剂量组大鼠肺泡炎和肺纤维化程度降低(P<0.01),RT-PCR和Western blot结果显示:TGFβ-1、Smad2、Smad3 mRNA的表达水平升高(P<0.05),α-SMA蛋白表达水平降低(P<0.05),PPAR-γ蛋白表达水平升高(P<0.05)。结论 TFSB对博莱霉素致大鼠肺纤维化有一定的防治作用,其机制可能与机体内的抗氧化和下调TGF-β1信号通路有关。     

Enzymes of collagen synthesis in lung tissues of bleomycin-induced pulmonary fibrosis

The activities of prolyl hydroxylase (Pro-OHase), galactosylhydroxylysyl glucosyltransferase (Glu-Gal-Hyl-Tase), and hydroxylysyl galactosyltransferase (Gal-Hyl-Tase) were assayed in lung tissues of hamsters with bleomycin-induced experimental pulmonary fibrosis. Serum Glu-Gal-Hyl-Tase and aspartate transaminase (Asp-NH₂-Tase) were measured in the same animals. Lung fibrosis was induced by intratracheal bleomycin instillation, and the enzyme activities were assayed 2, 3, and 4 weeks after bleomycin administration. The activities of the three lung enzymes increased significantly after bleomycin instillation. However, no difference in the values of serum Glu-Gal-Hyl-Tase or Asp-NH₂-Tase were observed. Histologic examination of lung sections indicated progressive fibrotic foci. These results thus indicate that the activities of collagen processing enzymes are elevated in the fibrotic lung tissues as a reflection of the increased rate of collagen synthesis during the period of active fibrogenesis, but unlike liver fibrosis, the elevation of tissue Glu-Gal-Hyl-Tase is not predicted by a corresponding increase in serum levels of this enzyme.     

Berberine regulates pulmonary inflammatory microenvironment and decreases collagen deposition in response to bleomycin-induced pulmonary fibrosis in mice

The purpose of this study was to explore the protective effect and potential mechanism of berberine on bleomycin (BLM)-induced fibrosis after lung injury in conjunction with network pharmacology. Berberine and pulmonary fibrosis prediction targets were collected for Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and so forth. A single intranasal dose of BLM (2.5 mg/kg) was administered to establish a model of fibrosis after lung injury, and berberine (50 mg/kg) was administered intraperitoneally daily for treatment. Network pharmacology results suggested that the mitogen-activated protein kinase (MAPK) signalling pathway may be a potential mechanism of berberine in delaying pulmonary fibrosis. The results of animal experiments showed that compared with the BLM group, after 14 days of berberine treatment, lung inflammatory cell aggregation was reduced and the expression levels of tumour necrosis factor-α (TNF-α), interleukin (IL)-8 and IL-6 were down-regulated in mice (p < 0.05); after 42 days of berberine treatment, the expression levels of transforming growth factor (TGF)-β1, platelet-derived growth factor-AB (PDGF-AB), hydroxyproline (HYP) and α-smooth muscle actin (α-SMA) were significantly down-regulated (p < 0.05), and the expression levels of total p38 MAPKα and p38 MAPKα (pT180/Y182) were down-regulated also (p < 0.05), inhibited collagen production and deposition, and increased the survival rate of mice to 70%. In conclusion, berberine attenuated inflammation mice, inhibited collagen production and showed some anti-pulmonary fibrosis potential in the MAPK signalling pathway.     

磷酸二酯酶4B在博来霉素诱导的肺纤维化小鼠肺组织中的表达与作用

目的观察博来霉素诱导的小鼠肺纤维化过程中磷酸二酯酶4B(PDE4B)mRNA和蛋白表达随时间的变化,初步探究PDE4B在小鼠肺纤维化过程中的作用。方法采用气道滴入博来霉素2.5 mg·kg-1制备肺纤维化模型,在造模后第3,7,14,21和28天进行支气管肺泡灌洗并留取肺组织。应用细胞形态学方法计数支气管肺泡灌洗液(BALF)中白细胞总数和分类;ELISA法测定BALF中巨噬细胞炎症蛋白2(MIP2)、白细胞介素6(IL-6)、IL-1β和转化生长因子β1(TGF-β1)含量;胶原试剂盒测定法测定肺组织胶原含量;髓过氧化物酶(MPO)活性测定试剂盒测定组织匀浆液中MPO活性;实时荧光定量PCR方法检测小鼠肺组织中PDE4B mRNA表达;免疫组织化学法检测PDE4B的分布。结果博来霉素气道滴入诱导的肺纤维化随时间持续加重,第28天最明显,可见肺组织明显实质化。BALF中白细胞总数、IL-1β和IL-6在造模后第3天达峰值,分别为正常对照组的22.0,2.0和2.8倍;MPO和TGF-β1在第7天达峰值,分别为正常对照组的1.9和5.5倍;胶原含量、MIP-2和PDE4B mRNA表达从造模后呈持续增长趋势,分别为正常对照组的1.6,2.7和2.6倍。免疫组织化学法检测结果表明,PDE4B分布于炎症细胞和纤维化的肺组织。结论PDE4B在肺纤维化中发挥重要作用,可能是一个特异性的药物作用靶点。     

Erdosteine prevents bleomycin-induced pulmonary fibrosis in rats

Oxidative stress plays an important role in the pathogenesis of idiopathic pulmonary fibrosis. Therefore, erdosteine, an antioxidant, is expected to have an inhibitor potential against the disease. Rats were given one dose of bleomycin in pulmonary fibrosis groups and saline in controls. The first dose of oral erdosteine (10 mg/kg/day) was given 2 days before the bleomycin injection to achieve the plateau level in blood and continued until killing. At day 14, fibrotic changes were evaluated, using Aschoft's criteria and lung hydroxyproline content. Bleomycin produced a fivefold increase in fibrosis score that was decreased by 87% by erdosteine (P>0.001) and almost twofold increases in hydroxyproline content which were completely prevented by erdosteine. Myeloperoxidase activities and MDA levels, which were significantly higher in the bleomycin group, were then significantly attenuated by erdosteine. These results revealed that oral erdosteine may prevent the development of acute pulmonary inflammation caused by bleomycin injection via the repression of neutrophil accumulation and lipid peroxidation, resulting in the inhibition of subsequent lung fibrosis.     

Feitai attenuates bleomycin-induced pulmonary fibrosis in rats

Pulmonary fibrosis is a common consequence of numerous pulmonary diseases. The current therapeutic approaches for this condition are unsatisfactory. Feitai, a composite formula consisting of several herbs, is used in China as a folk remedy for treating patients with pulmonary tuberculosis. In this study, we extensively investigate the effects and mechanisms of Feitai on bleomycin (BLM)-induced pulmonary fibrosis in rats. One hundred and twenty male Sprague-Dawley rats were randomly divided into four groups, referred to as the saline-water, saline-Feitai, BLM-water, and BLM-Feitai groups. Following a single instillation of BLM (5 mg/kg) or saline, rats were orally administered Feitai at a dose of 3 g/kg body weight or sterilized distilled water once daily. Rats were killed at 7, 14, or 28 d post-BLM. Inflammatory cell count, protein concentration, and lactate dehydrogenase activity in bronchoalveolar lavage fluid were measured, and myeloperoxidase activity and lipid peroxide content in lung homogenates were analyzed. Treatment with Feitai inhibited lung fibrotic progression induced by BLM, as indicated by the decrease in lung hydroproline content and lung fibrosis score at 28 d post-BLM. This was accompanied by significant amelioration of BLM-induced body weight loss, lung edema, and inflammatory response during the development of lung injury in the acute phase. The results strongly indicate the beneficial effects of Feitai in protecting against BLM-induced pulmonary fibrosis. Furthermore, the inflammatory response and lipid peroxidation were inhibited by Feitai, suggesting that the effect of this formula on BLM-induced lung injury and fibrosis is associated with antiinflammatory and antioxidant properties.     

Curcumin inhibition of bleomycin-induced pulmonary fibrosis in rats

Curcumin, an anti-inflammatory, antioxidant, was evaluated for its ability to suppress bleomycin (BLM)-induced pulmonary fibrosis in rats. A single intratracheal instillation of BLM (0.75 U 100(-1) g, sacrificed 3, 5, 7, 14 and 28 days post-BLM) resulted in significant increases in total cell numbers, total protein, and angiotensin-converting enzyme (ACE), and alkaline phosphatase (AKP) activities in bronchoalveolar lavage fluid. Animals with fibrosis had a significant increase in lung hydroxyproline content. Alveolar macrophages from BLM-administered rats elaborated significant increases in tumour necrosis factor (TNF)-alpha release, and superoxide and nitric oxide production in culture medium. Interestingly, oral administration of curcumin (300 mg kg(-1) 10 days before and daily thereafter throughout the experimental time period) inhibited BLM-induced increases in total cell counts and biomarkers of inflammatory responses in BALF. In addition, curcumin significantly reduced the total lung hydroxyproline in BLM rats. Furthermore, curcumin remarkably suppressed the BLM-induced alveolar macrophage production of TNF-alpha, superoxide and nitric oxide. These findings suggest curcumin as a potent anti-inflammatory and anti-fibrotic agent against BLM-induced pulmonary fibrosis in rats.     

卡托普利对博来霉素诱导大鼠肺纤维化的影响

目的探讨博来霉素（bleomycin）中毒肺BLM纤维化机制及卡托普利对BLM中毒肺纤维化的干预作用。方法 60只SD大鼠随机分为3组：正常对照（control）组、单纯BLM染毒（BLM）组和BLM染毒卡托普利（captopril,CPT）治疗（BLM＋CPT）组。14 d后处死各组动物,记录肺系数;病理组织学检查。结果 BLM染毒后大鼠肺系数增大,肺泡炎、肺纤维化程度积分明显增高。CPT均明显减轻了大鼠肺损伤、肺纤维化程度。结论 CPT对BLM中毒大鼠肺损伤、肺纤维化程度有一定改善作用,可能与其抑制血管紧张素Ⅱ有关。