Changes in pulmonary surfactant function and composition in bleomycin-induced pneumonitis and fibrosis

Bleomycin is a widely accepted cancer drug but may induce life-threatening interstitial lung disease in a subset of patients. We evaluated the effect of bleomycin administration on pulmonary surfactant function and composition in rabbit lungs. In order to obtain a uniform response to bleomycin, aerosol technology was employed for bronchoalveolar delivery of 1.8 U/kg b.w. bleomycin. On days 4, 8, 16, 24, 32, and 64 after challenge, bronchoalveolar lavages were performed. Sham-aerosolized rabbits served as controls. In the early acute respiratory distress syndrome (ARDS)-like post-bleomycin period (4-16 days), marked loss of surface activity of the large surfactant aggregate (LA) fraction of surfactant was noted. In parallel, reduced percentages of LA, but only minor changes in surfactant apoproteins (SP)-A, SP-B, and SP-C, were encountered. Analysis of the surfactant lipid profile showed impressively enhanced cholesterol and significantly decreased phosphatidylglycerol (PG) levels. The relative content of dipalmitoyl-PC (DPPC) was slightly increased, and a several-fold increase within the 1-O-alkyl-2-acyl subclass of PC was observed. During the prolonged fibroproliferative period, a highly significant downregulation of SP-B and SP-C levels was observed. This was paralleled by an upregulation of the total extracellular phospholipid pool, with a far-reaching normalization of the (phospho)-lipid profile. The biophysical surfactant function never fully normalized within the 64-day observation period. In conclusion, bleomycin caused marked abnormalities of pulmonary surfactant, with the profile of changes being different between the early ARDS and the late fibrotic phase.     

Inhibitory effect of emodin on bleomycin-induced pulmonary fibrosis in mice

1. Currently, there is no satisfactory treatment for pulmonary fibrosis. Emodin, a component in Chinese herbs, has been shown to have an antifibrotic effect on pancreatic fibrosis and liver fibrosis. In the present study, we tested the hypothesis that emodin may attenuate the development of pulmonary fibrosis. 2. Mice were randomly divided into five groups (n = 16 in each). One group was a control group; the remaining four groups were treated with intratracheal instillation of 3 mg/kg bleomycin (BLM). The following day, emodin (5, 10 or 20 mg/kg per day, p.o.) treatment was started for three of the BLM-treated groups and was continued for 21 days. The fourth BLM-treated group (and the control group) received daily 0.5% sodium carboxymethyl cellulose (placebo) by gavage over the same period. 3. Bleomycin challenge provoked severe pulmonary fibrosis, with marked increases in fibrosis fraction, hydroxyproline content and myeloperoxidase activity in lung tissue. Emodin treatment (10 and 20 mg/kg per day, p.o.) attenuated all these biochemical indices, as well as histopathological alterations induced by BLM. Furthermore, in mice injected with BLM, elevated levels of transforming growth factor-beta1, interleukin (IL)-4 and IL-13 were found in bronchoalveolar lavage fluid. These increases were significantly inhibited by 10 and 20 mg/kg per day emodin. 4. In cell culture, exposure of cells to 6.25, 12.5, 25 or 50 micromol/L emodin for 24 h decreased fibroblast proliferation. Treatment of cells with the same concentrations of emodin for 72 h decreased collagen production by fibroblasts. In addition, emodin (6.25, 12.5, 25 or 50 micromol/L) inhibited the steady state expression of alpha1 (I) procollagen and alpha2 (I) procollagen mRNA in a dose-dependent manner. 5. The results of the present study suggest that emodin may be effective in the treatment of pulmonary fibrosis.     

Effect of antibody against integrin α4 on bleomycin-induced pulmonary fibrosis in mice

Integrins are a family of transmembrane glycoproteins that can interact with components of the extracellular matrix. The α4β1 and α4β7 integrins are heterodimeric leukocyte cell surface molecules critical to their cell and matrix adhesive interactions. Evidence for a central role for the α4 integrins in leukocyte pathophysiology in the lung is well documented. In this study, we tested the hypothesis that neutralizing antibody for integrin α4 (PS2) may reduce bleomycin (BL)-induced lung fibrosis in vivo. Male C57BL/6 mice were injected intratracheally with saline (SA) or BL (0.08 U/mouse) followed by intraperitoneal injection of SA, isotype control antibody (1E6), or PS2 (100 μg) three times a week. Twenty-one days after the intratracheal instillation, mice were killed for bronchoalveolar lavage (BAL), biochemical, histopathological, and immunohistological analyses. Treatment with PS2 significantly reduced BL-induced increases in lung lipid peroxidation and hydroxyproline content. Lung histopathology also showed reduced fibrotic lesions in the BL-treated lungs by treatment with PS2. BL-treated mouse lungs also showed induction of cells with the myofibroblast phenotype, as indicated by the increased expression of α-smooth muscle actin (αSMA), whereas treatment with PS2 minimized the BL-induced αSMA expression. Furthermore, treatment with PS2 reduced the BL-induced increase in the BAL total cell number, and attenuated the BL-induced increase in the BAL protein level. It is concluded that integrin α4 may play an important role in BL-induced pulmonary fibrosis, and the use of anti-α4 antibody offers therapeutic antifibrotic potential in vivo.     

丹参对博莱霉素所致大鼠肺纤维化保护作用的实验研究

目的:探讨丹参对博莱霉素所致大鼠肺纤维化的保护作用及其作用机理.方法:通过气管内注射博莱霉素复制大鼠肺纤维化模型,在造模后的d 1和d 7开始给予丹参注射液治疗,分别在d 7、d 28处死大鼠,对肺组织匀浆中的脂质过氧化物(MDA)、谷胱甘肽(GSH)、羟脯氨酸(HYP)和肺组织病理学变化测量.结果:在造模后d 1给予丹参注射液治疗能抑制实验性肺纤维化大鼠的肺组织匀浆中的MDA、HYP异常升高和GSH的下降,减轻其肺部的病理损害;在造模后d 7给予丹参注射液治疗,丹参治疗组与模型组上述观察指标比较无统计学差异.结论:丹参注射液可预防大鼠肺纤维化,对已形成的肺纤维化无逆转作用.     

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。     

吲哚-3-原醇对博莱霉素所致小鼠肺纤维化的干预作用及其机制

目的观察吲哚-3-原醇(indole-3-carbinol,I3C)对博莱霉素致小鼠肺纤维化的干预作用并初步探讨其机制。方法Imprinting Control Region(ICR)小鼠随机分为6组:正常组、模型组、醋酸泼尼松组(6.67 mg.kg-1)、吲哚-3-原醇小、中、大剂量组(25、50、100 mg.kg-1)。小鼠气管内注射博莱霉素制备肺纤维化模型后,第2天给予相应药物,每天1次。连续给药28 d后处死小鼠检测其肺系数、血清和肺组织羟脯氨酸(Hydroxyproline,HYP)、血清总抗氧化能力(To-tal antioxidative capacity,T-AOC);取固定部位肺组织切片HE染色,进行病理学观察;RT-PCR法检测肺组织中α-SMA、CollagenⅠ、TGF-β、Smad2 mRNA的表达;Western blot法检测肺组织中α-SMA、TGF-β、Smad2蛋白的表达。结果吲哚-3-原醇能够提高小鼠血清的总抗氧化能力(P<0.05或P<0.01),同时可以降低肺系数和肺组织中的HYP含量(P<0.05或P<0.01);光镜观察表明吲哚-3-原醇能明显减轻肺纤维化小鼠肺泡炎和肺纤维化程度(P<0.05或P<0.01);RT-PCR检测发现吲哚-3-原醇能明显降低肺组织α-SMA、CollagenⅠ、TGF-β、Smad2基因表达水平(P<0.05或P<0.01);Western blot分析结果说明吲哚-3-原醇能明显降低肺组织α-SMA、TGF-β、Smad2蛋白表达(P<0.05或P<0.01)。结论吲哚-3-原醇可能通过抑制TGF-β/Smad信号通路来减轻博莱霉素致小鼠肺纤维化。     

Acute exposure of mice to hydrochloric acid leads to the development of chronic lung injury and pulmonary fibrosis

Objective: Accidental exposure to hydrochloric acid (HCl) is associated with acute lung injury in humans, development of long-term chronic airway obstruction, and fibrosis. However, the mechanisms responsible for the progression to pulmonary fibrosis remain unclear. We utilized a mouse model of progressive lung injury from a single exposure to HCl to investigate the effects of HCl on the lower respiratory tract.

Materials and methods: HCl (0.05–0.3?N) or saline was injected intratracheally into male C57Bl/6J mice. At 1, 4, 10 and 30 days post instillation, bronchoalveolar lavage fluid (BALF) and lung tissues were collected and examined for multiple outcomes.

Results and discussion: We observed an early inflammatory response and a late mild inflammation present even at 30?d post HCl exposure. Mice treated with HCl exhibited higher total leukocyte and protein levels in the BALF compared to the vehicle group. This was characterized by increased number of neutrophils, monocytes, and lymphocytes as well as pro-inflammatory cytokines during the first 4?d of injury. The late inflammatory response exhibited a predominant presence of mononuclear cells, increased permeability to protein, and higher levels of the pro-fibrotic mediator TGFβ. Pro-fibrotic protein biomarkers, phosphorylated ERK, and HSP90, were also overexpressed at 10 and 30?d following HCl exposure. In vivo lung function measurements demonstrated lung dysfunction and chronic lung injury associated with increased lung hydroxyproline content and increased expression of extracellular matrix (ECM) proteins. The acute inflammation and severity of fibrosis increased in HCl-concentration dependent manner.

Conclusions: Our findings suggest that the initial inflammatory response and pro-fibrotic biomarker upregulation may be linked to the progression of pulmonary fibrosis and airway dysfunction and may represent valuable therapeutic targets.     

川芎嗪对内毒素诱导的急性肺损伤大鼠肺纤维化和炎症反应的调节作用

目的：探索川芎嗪对内毒素诱导的急性肺损伤大鼠肺纤维化和炎症反应的影响及其机制。方法：大鼠随机分为5组：对照组、LPS模型组、LPS+川芎嗪（50,100,200 mg·kg^-1）组。苏木素伊红（HE）染色观察肺组织病变。Masson染色分析肺纤维化。酶联免疫吸附实验（ELISA）检测白细胞介素（interleukin,IL）-6,IL-1β,肿瘤坏死因子（tTNF-α）和IL-18水平。蛋白印记检测α平滑肌肌动蛋白（α-SMA）,转化生长因子β1（TGF-β1）,Ⅰ型胶原蛋白（collagen type Ⅰ）,Ⅲ型胶原蛋白（collagen type Ⅲ）,PI3K,P-PI3K,AKT,P-AKT,mTOR和P-mTOR的蛋白水平。结果：LPS模型组大鼠肺组织湿/干重之比高于对照组（P<0.01）。与LPS模型组相比,LPS+川芎嗪（100,200 mg·kg^-1）组大鼠肺组织湿/干重之比降低（P<0.01）。川芎嗪处理可改善LPS模型组大鼠肺组织病变及纤维化。LPS模型组大鼠α-SMA,TGF-β1,collagen type Ⅰ和collagen type Ⅲ表达高于对照组（P<0.01）。与LPS模型组相比,LPS+川芎嗪（50,100,200 mg·kg^-1）组大鼠α-SMA、TGF-β1、collagen type Ⅰ和collagen type Ⅲ表达降低（P<0.01）。与对照组相比,LPS模型组大鼠IL-6、IL-1β、IL-18和TNF-α水平升高（P<0.01）。LPS+川芎嗪（50,100,200 mg·kg^-1）组大鼠IL-6、IL-1β、IL-18和TNF-α水平低于LPS模型组（P<0.01）。LPS模型组大鼠P-PI3K/PI3K,P-AKT/AKT和P-mTOR/mTOR比值高于对照组（P<0.01）。与LPS模型组相比,LPS+川芎嗪（50,100,200 mg·kg^-1）组大鼠P-PI3K/PI3K,P-AKT/AKT和P-mTOR/mTOR比值下降（P<0.01）。结论：川芎嗪可通过抑制PI3K/AKT/mTOR信号通路活化减轻内毒素诱导的急性肺损伤大鼠肺纤维化和炎症反应。     

Heat shock protein 70 protects against bleomycin-induced pulmonary fibrosis in mice

Idiopathic pulmonary fibrosis (IPF) involves infiltration of leucocytes, pulmonary injury, fibrosis and resulting pulmonary dysfunction. Myofibroblasts and transforming growth factor (TGF)-β1 have been suggested to play a major role in the pathology and the myofibroblasts are derived from both lung epithelial cells through epithelial-mesenchymal transition (EMT) and activation of lung fibroblasts. Heat shock protein 70 (HSP70) confers protection against various stressors and has the anti-inflammatory activity. In this study, we examined the effect of expression of HSP70 on bleomycin-induced pulmonary fibrosis in mice, a tentative animal model of IPF. Bleomycin-induced pulmonary injury and inflammatory response were ameliorated in transgenic mice overexpressing HSP70 compared to wild-type mice, even though bleomycin-induced pulmonary fibrosis and dysfunction were also suppressed in the transgenic mice. The production of TGF-β1 and expression of pro-inflammatory cytokines was lower in cells from the transgenic mice than wild-type mice after the administration of bleomycin. In vitro, the suppression of HSP70 expression stimulated TGF-β1-induced EMT-like phenotypes of epithelial cells but did not affect the TGF-β1-dependent activation of fibroblasts. Orally administered geranylgeranylacetone (GGA), a clinically used drug with HSP-inducing activity, conferred protection against bleomycin-induced pulmonary injury, as well as against the inflammatory response, fibrosis and dysfunction. These results suggest that HSP70 plays a protective role against bleomycin-induced pulmonary injury, inflammation, fibrosis and dysfunction through cytoprotective effects and by inhibiting the production of TGF-β1, TGF-β1-dependent EMT of epithelial cells and expression of pro-inflammatory cytokines. Results also suggest that HSP70-inducing drugs, such as GGA, could be beneficial in the prophylaxis of IPF.     

芹菜素对百草枯致小鼠肺纤维化保护作用及机制研究

目的 研究芹菜素对百草枯致小鼠肺纤维化的保护作用及机制。方法 将小鼠随机分为对照组、模型组、地塞米松组（1 mg/kg）和芹菜素低、高剂量（10、20 mg/kg）组,除对照组外,其他各组ip百草枯（50 mg/kg）建立小鼠肺纤维化模型,第8天开始ig给药,连续给药21 d。观察小鼠体质量变化;测量小鼠肺系数;采用HE染色观察肺组织形态学变化;免疫组化法观察小鼠肺组织中Ⅰ、Ⅲ型胶原表达变化;ELISA法检测各组小鼠肺组织中Ⅰ、Ⅲ型胶原和炎症因子肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6、IL-1β含量;Western blotting法测定肺组织Caveolin-1蛋白表达水平。结果 造模第28天时,与模型组比较,芹菜素低、高剂量组小鼠体质量显著增加（P<0.05）;地塞米松组和芹菜素低、高剂量组肺系数显著降低（P<0.05）;芹菜素低、高剂量组肺组织内炎性细胞浸润较少、肺泡壁断裂及肺泡腔融合较少,肺纤维化程度较轻;芹菜素低、高剂量组和地塞米松组小鼠的肺组织中Ⅰ、Ⅲ型胶原和IL-6、IL-1β、TNF-α含量显著下降（P<0.01）,Caveolin-1蛋白表达显著下降（P<0.01）。结论 芹菜素对百草枯诱导小鼠肺纤维化具有保护作用,其机制可能与Ⅰ、Ⅲ型胶原含量及炎症因子水平下调、Caveolin-1蛋白表达水平下调相关。     

白藜芦醇对博莱霉素诱导的小鼠肺纤维化的作用

目的:探讨白藜芦醇(Res)对博莱霉素(BLM)致肺纤维化(PF)的治疗作用.方法:复制小鼠博莱霉素致PF模型,采用光镜观察组织学改变.测定肺组织羟脯氨酸(HyP)、超氧化物歧化酶(SOD)、丙二醛(MDA)以反映肺细胞损伤及PF的程度.结果:Res能显著降低实验性PF小鼠肺组织中Hyp、MDA的含量,显著提高小鼠肺组织的SOD活力.病理组织学检查亦表明,Rea能明显改善肺泡炎症程度和PF程度.结论:Res对实验性小鼠PF具有一定的治疗作用.     

异莲心碱对百草枯诱导的小鼠急性肺损伤及肺纤维化的保护作用(英文)

目的探讨异莲心碱对百草枯(PQ)诱导的小鼠急性肺损伤及肺纤维化是否有保护作用。方法一次性给不同剂量PQ分别制备小鼠急性肺损伤模型(45mg·kg-1,ip)和肺纤维化模型(100mg·kg-1,ig)。异莲心碱在给PQ前1 d开始给予至实验结束。实验分空白对照组、模型组(PQ)、单用异莲心碱组及异莲心碱治疗组(异莲心碱+PQ)。急性肺损伤模型组给予PQ后第8,24及48 h观察异莲心碱(20 mg.kg-1,ig,每天3次)对血浆及支气管肺泡灌洗液(BALF)中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量及碱性磷酸酶(ALP)活性的影响。肺纤维化组给予PQ后14 d观察异莲心碱(10,20, 40 mg·kg-1,ig,每天2次)对肺组织羟脯氨酸含量、肺组织中转化生长因子β1(TGF-β1)及基质金属蛋白酶2(MMP-2)表达的影响。同时采用HE染色方法观察两模型在上述各时间点的肺组织病理变化。结果在急性肺损伤模型中,与空白组比较,模型组的肺组织出现充血、出血、炎性渗出及水肿等病变,血浆和BALF中的SOD活性明显下降,MDA水平明显升高,而血浆中的ALP活性显著升高;与模型组比较,异莲心碱治疗组的肺组织炎症反应减轻,血浆和BALF中SOD活性明显升高,血浆中ALP活性以及血浆和BALF中MDA含量明显降低;单用异莲心碱以上指标无明显作用。在肺纤维化模型中,一次性给PQ14 d后,与空白组比较,模型组肺组织出现肺间质增厚和胶原纤维增生等病理变化,肺组织中羟脯氨酸含量明显升高(2.44±0.33 )vs(1.26±0.10 )mg·g-1湿组织;与模型组比较,异莲心碱10, 20和40mg·kg-1治疗组间质炎症及肺纤维化病变有所改善,肺组织中羟脯氨酸含量显著减少分别为(2.11±0.21), (1.94±0.24)和(1.89±0.26)mg·g-1湿组织;与模型组比较,异莲心碱40mg·kg-1治疗组肺组织中TGF-β1和MMP-2的表达明显降低。结论异莲心碱对PQ诱导的急性肺损伤及肺纤维化具有一定的保护作用。     

芒果三芪肺纤方对博莱霉素致小鼠肺纤维化作用研究

目的 观察芒果三芪肺纤方对博莱霉素诱导小鼠肺纤维化的改善作用。方法 昆明小鼠随机分为对照组,模型组,地塞米松（阳性药,1 mg/kg）组,芒果三芪肺纤方高、中、低剂量（以生药计5.00、3.30、1.65 g/kg）组,通过鼻腔1次性滴入6 mg/kg盐酸博莱霉素制备小鼠肺纤维化模型,对照组滴入等体积的生理盐水。于造模后第14天开始ig给药,每天给药1次,连续给药14 d后处死小鼠,试剂盒法测定肺组织匀浆中的羟脯氨酸（HYP）、丙二醛（MDA）、白介素-1β（IL-1β）水平和超氧化物岐化酶（SOD）活力;HE染色观察肺组织病理切片,Masson染色观察胶原纤维的分布情况。结果 与模型组比较,芒果三芪肺纤方高、中、低剂量组小鼠HYP和IL-1β水平均显著降低（P<0.01）,SOD活力显著提高（P<0.05、0.01）,高和中剂量组小鼠MDA水平显著降低（P<0.05、0.01）;HE和Masson染色显示,芒果三芪肺纤方组肺泡炎损伤和肺纤维化病变程度明显减轻（P<0.01）,高剂量组效果最好。结论 芒果三芪肺纤方通过提高SOD活力、降低MDA水平,发挥抗自由基损伤作用;降低HYP的水平,减少纤维蛋白的形成;降低IL-1β水平,发挥抗炎作用;显著减轻肺泡炎症损伤、抑制肺纤维化病变程度,对博莱霉素诱导的小鼠肺纤维化具有很好的改善作用。     

沙苑子总黄酮对博莱霉素致大鼠肺纤维化的干预作用及其机制研究

目的探讨沙苑子总黄酮(FAC)对博莱霉素(BLM)所致实验性大鼠肺纤维化的干预作用,并探讨其作用机制。方法采用气管内滴注博莱霉素法制作大鼠肺纤维化模型,并随机分为6组。给予沙苑子总黄酮灌胃(ig)治疗28 d后,腹主动脉放血法处死动物,测定血清和总抗氧化能力(T-AOC)及肺组织中羟脯氨酸(HYP)含量;ELISA的方法检测实验大鼠肺泡灌洗液中IL-1β和IL-6的含量;取固定部位肺组织作HE及Masson染色,进行病理组织学观察;电镜观察大鼠肺组织的超微结构变化;Westen blot法检测沙苑子总黄酮TGF-β/Smad信号通路相关蛋白的表达。结果与模型组相比,沙苑子总黄酮各组大鼠肺组织中HYP含量明显降低(P<0.01或P<0.05),且有剂量依赖趋势;大鼠血清及肺组织中T-AOC增强(P<0.01或P<0.05);肺泡灌洗液中细胞因子IL-1β及IL-6的含量明显降低(P<0.01或P<0.05);组织病理学观察肺泡炎症(P<0.01或P<0.05)及肺组织纤维化(P<0.01或P<0.05)的程度明显减轻;West-ern blot检测结果显示TGF-β/Smad信号通路相关蛋白TGF-β、Smad2、α-SMA含量均明显降低。结论沙苑子总黄酮对博莱霉素致大鼠肺纤维化病变有一定的治疗作用,其作用机制可能与其抑制炎症细胞因子的活化、抗氧化作用、抑制胶原的形成以及调控TGF-β/Smad信号通路有关。     

灯盏花素对博来霉素诱导小鼠肺纤维化的保护作用

目的观察灯盏花素对肺纤维化的保护作用,探讨其可能的作用机制。方法观察灯盏花素对体外培养小鼠胚肺成纤维细胞L929增殖、激活及细胞外基质分泌的影响;应用博来霉素诱导小鼠肺纤维化模型,观察灯盏花素对肺纤维化的保护作用。结果灯盏花素体外对小鼠胚肺成纤维细胞L929无直接细胞毒作用,但能抑制TGF-β1促进的增殖、激活及层粘连蛋白(LN)、Ⅰ型胶原(ColⅠ)分泌。体内灯盏花素能抑制博来霉素诱导的小鼠血清TGF-β1升高,阻止博来霉素诱导的小鼠肺脏SOD、POD、CAT降低,并降低肺脏羟脯氨酸、胶原、MDA及TGF-β1含量。结论灯盏花素有肺纤维化保护作用,其机制可能是通过增加抗氧化防御系统和阻止TGF-β信号实现的。     

Effect of Feixian Recipe on laminin, collagen I and III in rats with pulmonary fibrosis induced by bleomycin

The aim of this paper is to observe the effect of the Feixian Recipe on pulmonary fibrosis in rats. A rat model with pulmonary fibrosis was established by intratracheal injection of bleomycin. On days 14, 28 and 45, the contents of laminin, collagen I and collagen III in lung tissue homogenate in the model group, the sham operated group, the Feixian group and the prednisone group were measured. The contents of laminin and collagen I and III were decreased significantly by the Feixian Recipe. Feixian Recipe has a significant therapeutic effect on bleomycin-induced pulmonary fibrosis in rats. __________ Translated from Journal of Beijing University of Traditional Chinese Medicine, 2007, 30(9): 608–610 [译自: 北京中医药大学学报]     

枇杷叶三萜酸对博来霉素致大鼠肺纤维化的干预作用

目的观察博来霉素(BLM)致大鼠肺纤维化模型的超微病理过程,肺组织匀浆MDA含量、SOD活性,血清iN-OS、eNOS、NO活性的动态变化及枇杷叶三萜酸(TAL)对其的影响。方法通过气管内一次性滴注博来霉素(5 mg.kg-1)复制大鼠肺纤维化模型。治疗组于造模后d 2开始灌胃给予150 mg.kg-1 TAL。各组大鼠于给药后d 7、14、28随机取样8只。采集血清,留取肺组织制备肺组织匀浆,分别测定匀浆中丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、血清中一氧化氮(NO)含量、诱生型及内皮型一氧化氮合酶(iNOS、eNOS)活性。制备电镜标本行电镜观察。结果电镜可见,模型7 d组大鼠Ⅰ、Ⅱ型肺泡上皮胞质水肿,线粒体肿胀,嵴空化。血管内皮细胞肿胀,肺泡隔胶原增多。肺间质及肺泡腔内炎性细胞增多。14、28 d BLM组Ⅰ型肺泡上皮胞质水肿更明显,Ⅱ型细胞较7 d BLM组减少,巨噬细胞凋亡增加,间质纤维组织增生明显。TAL能明显改善模型组大鼠肺脏组织结构,减轻肺纤维化增生程度。与正常组相比,模型组各时间段血清iNOS活性、肺匀浆MDA含量均升高,而eNOS活性下降(P<0.01 or P<0.05);模型7、14 d血清NO含量增加,肺组织匀浆SOD活性下降(P<0.01 or P<0.05),模型组28 d恢复正常。TAL给药可降低各时段增高的iNOS、NO及MDA含量,同时升高SOD及eNOS活性(P<0.01 or P<0.05)。结论博来霉素诱发大鼠肺纤维化早期以肺泡炎为主,28 d肺纤维化为主要病变。肺内一氧化氮合酶的含量变化,NO的大量生成及过氧化可能是促使肺纤维化形成的重要因素,TAL对肺纤维化的防治作用可能与其调节iNOS、eNOS比例、抗脂质过氧化有关。     

Berberine ameliorates renal interstitial inflammation and fibrosis in mice with unilateral ureteral obstruction

Berberine acts via multiple pathways to alleviate fibrosis in various tissues and shows renoprotective effects. However, its role and underlying mechanisms in renal fibrosis remain unclear. Herein, we aimed to investigate the protective effects and molecular mechanisms of berberine against unilateral ureteric obstruction-induced renal fibrosis. The results indicated that berberine treatment (50 mg/kg/day) markedly alleviated histopathological alterations, collagen deposition and inflammatory cell infiltration in kidney tissue and restored mouse renal function. Mechanistically, berberine intervention inhibited NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome activation and the levels of the inflammatory cytokine IL-1β in the kidneys of unilateral ureteric obstruction mice. In addition, berberine relieved unilateral ureteric obstruction-induced renal injury by activating adenosine monophosphate-activated protein kinase (AMPK) signalling and promoting fatty acid β-oxidation. In vitro models showed that berberine treatment prevented the TGF-β1-induced profibrotic phenotype of hexokinase 2 (HK-2) cells, characterized by loss of an epithelial phenotype (alpha smooth muscle actin [α-SMA]) and acquisition of mesenchymal marker expression (E-cadherin), by restoring abnormal fatty acid β-oxidation and upregulating the expression of the fatty acid β-oxidation related-key enzymes or regulators (phosphorylated-AMPK, peroxisome proliferator activated receptor alpha [PPARα] and carnitine palmitoyltransferase 1A [CPT1A]). Collectively, berberine alleviated renal fibrosis by inhibiting NLRP3 inflammasome activation and protected tubular epithelial cells by reversing defective fatty acid β-oxidation. Our findings might be exploited clinically to provide a potential novel therapeutic strategy for renal fibrosis.     

Th 细胞亚群在肺纤维化小鼠中的动态表达

目的：研究辅助性T细胞亚群（Th17、Th1和Th2）在肺纤维化模型小鼠的动态表达。方法40只雄性C57／BL6小鼠随机均分为两组,分别经气管一次性注入生理盐水（A组）或博来霉素溶液0.04ml（5mg／kg,B组）,于第3、7、14和28天处死。采用HE和Masson染色评价小鼠肺组织病理形态变化;检测肺组织中羟脯氨酸（HYP）含量;应用流式细胞术分析脾脏中Th1、Th2和Th17细胞的动态变化。结果A组小鼠肺泡形态正常,肺组织HYP含量无明显变化。B组小鼠第7天时即有部分肺泡结构破坏,肺组织可见大量炎性细胞及红细胞浸润,伴细束状胶原纤维增生;第7、14、28天肺组织HYP含量高于A组（P＜0．05）;与A组比较,B组4个时间点的Th17和Th2细胞数增加,Th1细胞数减少（P＜0．05）。结论Th17、Th2和Th1细胞与特发性肺纤维化密切相关,可能参与其发病机制。