BACKGROUNDBleomycin-induced fibrosis is extensively used to model aspects of the pathogenesis of interstitial pulmonary fibrosis. This study aimed to determine the benefic effects and mechanisms of simvastatin on bleomycin-induced pulmonary fibrosis in mice.

METHODSBleomycin-induced pulmonary fibrosis mice were administered with simvastatin in different doses for 28 days. We measured inflammatory response, fibrogenic cytokines and profibrogenic markers in both bleomycin-stimulated and control lungs, and correlated these parameters with pulmonary fibrosis.

RESULTSSimvastatin attenuated the histopathological change of bleomycin-induced pulmonary fibrosis and prevented the increase of lung hydroxyproline content and collagen (I and III) mRNA expression induced by bleomycin. Moreover, simvastatin down-regulated the increased expression of transforming growth factor-beta1 (TGF-beta1) and connective tissue growth factor (CTGF) induced by bleomycin at both gene and protein levels. Simultaneously, the accumulation of neutrophils and lymphocytes and the increased production of tumor necrosis factor-alpha (TNF-alpha) in bronchial alveolar lavage fluid were inhibited by simvastatin in early inflammatory phase after bleomycin infusion. The higher dose of simvastatin was associated with a more significant reduction in these inflammatory and fibrotic parameters. Furthermore, the inactivation of p38, RhoA and Smad2/3 signaling pathways was observed during simvastatin administration.

CONCLUSIONSSimvastatin attenuated bleomycin-induced pulmonary fibrosis, as indicated by decreases in Ashcroft score and lung collagen accumulation. The inhibitory effect of simvastatin on the progression of pulmonary fibrosis may be demonstrated by reducing inflammatory response and production of TGF-beta1 and CTGF. These findings indicate that simvastatin may be used in the treatment of pulmonary fibrosis.

Animals ; Antibiotics, Antineoplastic ; Bleomycin ; Mice ; Mice, Inbred C57BL ; Pulmonary Fibrosis ; chemically induced ; metabolism ; pathology ; Simvastatin ; pharmacology

OBJECTIVETo evaluate effect of amygdalin on expression of four biomarkers in the animal model of pulmonary fibrosis induced by bleomycin.

METHODSRats were given one dose (5 mg/kg) of bleomycin in bleomycin-treated groups, amygdalin-treated groups and saline in controls by intratracheal instillation exposed surgically. The amygdalin-treated groups rats were treated with intraperitoneal injection of amygdalin (15 mg x kg(-1) x day(-1)). The rats were sacrificed 7, 14 and 28 days after bleomycin administration. Polarized light microscopy and Image-Pro Plus detected I and III collagen expressed in Paraffin-embedded lung sections stained with Sirius red. Surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF MS) with weak cationic proteinchip (CM10) detected differentially expressed proteins in the pooled serum samples of all groups.

RESULTSConsistent fibrotic responses were found in all bleomycin and amygdalin-tread groups. On the 7th, 14th and 28th day after bleomycin or saline instillation, four differentially expressed proteins were detected in the pooled serum of all groups rats, consisting of 4 proteins with mass/charge ratio of 3530.7, 7043.5, 8332.6 and 9068.0, respectively. Compared with control groups, protein peaks intensity ratio with mass/charge ratio of 3530.7 on 7, 28 d and 7043.5, 8332.6 and 9068.0 on 7, 14 and 28 d was > 2 in bleomycin-treated groups. Compared with amygdalin-treated groups, protein peaks intensity with mass/charge ratio of 3530.7 at 7, 14, 28 d had no change almost, but protein peaks intensity ratio with mass/charge ratio of 7043.5 at 7 d, 8332.6 on 28 d and 9068.0 on 14 d was > 2 in bleomycin-tread groups. All the four protein peaks intensity had no change almost at other point.

CONCLUSIONAmygdalin may reduce the bleomycin-induced increase of differentially expressed protein peak intensities in rat serum.

Amygdalin ; pharmacology ; Animals ; Biomarkers ; blood ; Bleomycin ; adverse effects ; Blood Proteins ; metabolism ; Male ; Pulmonary Fibrosis ; blood ; chemically induced ; Rats ; Rats, Wistar

Animals ; Bleomycin ; Epithelial-Mesenchymal Transition ; Lung ; Male ; Pulmonary Fibrosis/drug therapy* ; Rats ; Rats, Sprague-Dawley ; Simvastatin/pharmacology*

OBJECTIVETo study the role of mitogen-activated protein kinase (MAPK) pathway in the action mechanism of PYM-induced KB cell apoptosis.

METHODSWestern blot analysis was used to investigate the expression of the mitogen-activated protein kinase.

RESULTSWhen treated with PYM in cultured KB cells, the extracellular signal-regulated kinase (ERKl/2) showed a dose-and time-dependent decreasing in phosphorylation status of these proteins through a western blot analysis, whereas protein levels of p38 MAPK remained unchanged.

CONCLUSIONSThe mitogen-activated protein kinase (MAPK) pathway may play an important ro1e in PYM-induced apoptosis of KB cells.

Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Bleomycin ; analogs & derivatives ; pharmacology ; Humans ; KB Cells ; Mitogen-Activated Protein Kinases ; physiology ; p38 Mitogen-Activated Protein Kinases

OBJECTIVEUsing monoclonal antibody (mAb) Fab' fragment to develop mAb immunoconjugates for cancer.

METHODSFab' fragment of mAb 3A5 was prepared by digestion of the antibody with pepsin and then reduced by dithiothreitol (DT), while Fab' fragment of mAb 3D6 was obtained by digestion of the antibody with ficin and subsequently reduced by beta-mercaptoethanol. The conjugation between Fab' fragment and pingyangmycin (PYM), an antitumor antibiotic, was mediated by dextran T-40. Immunoreactivity of Fab'-PYM conjugates with cancer cells was determined by ELISA, and the cytotoxicity of those conjugates to cancer cells was determined by clonogenic assay. Antitumor effects of the Fab'-PYM conjugates were evaluated by subcutaneously transplanted tumors in mice.

RESULTSThe molecular weight of Fab' fragment was approximately 53 kD, while the average molecular weight of Fab'-PYM conjugate was 170 kD. The Fab'-PYM conjugates showed immunoreactivity with antigen-relevant cancer cells and selective cytotoxicity against target cells. Administered intravenously, Fab'-PYM conjugates were more effective against the growth of tumors in mice than free PYM and PYM conjugated with intact mAb.

CONCLUSIONFab'-PYM conjugate may be capable of targeting cancer cells and effectively inhibiting tumor growth, suggesting its therapeutic potential in cancer treatment.

Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antibodies, Monoclonal ; pharmacology ; Bleomycin ; analogs & derivatives ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Colonic Neoplasms ; pathology ; Female ; HT29 Cells ; Humans ; Immunoglobulin Fab Fragments ; pharmacology ; Immunotoxins ; pharmacology ; KB Cells ; Liver Neoplasms ; pathology ; Mice ; Tumor Cells, Cultured ; drug effects

OBJECTIVEThe purpose of this study was investigate the effect of hyperthemia on multidrug resistance in K562/ADM cell.

METHODSThe MDR1 (mulitdrug resistance gene) and MRP (multidrug resistant associated gene) gene expressions in Tca8113 and K562/ADM cell lines were analyzed by RT-PCR after treated with different cytotoxic drugs and different temperature (37 degrees C and 41 degrees C). The function and expression of Pgp and MRP were detected by fluorescence photometeric assays.

RESULTSInhibition rate of both cells was significantly enhanced by exposure to chemotherapeutic drugs and 41 degrees C temperature; Exposing to 41 degrees C hyperthemia reduced MDR1 and MRP expression and enhanced intracellular drug concentration as well in K562/ADM.

CONCLUSION41 degrees C hyperthemia could effectively enhance the inhibition rate of chemotherapeutic drugs and partially reverse the multidrug resistance. It is suggested that hyperthemia could be used as a method to overcome multidrug resistance.

Antineoplastic Agents ; pharmacology ; Bleomycin ; analogs & derivatives ; pharmacology ; Cisplatin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; genetics ; Genes, MDR ; genetics ; Humans ; Hyperthermia, Induced ; K562 Cells ; Methotrexate ; pharmacology ; Multidrug Resistance-Associated Proteins ; biosynthesis ; genetics ; Tongue Neoplasms ; genetics ; pathology

OBJECTIVETo study the combined damage-effects of low-intensity 2,450 MHz microwave (MW) with three chemical mutagens on human lymphocyte DNA.

METHODSDNA damage of lymphocytes exposed to microwave and(or) with chemical mutagens were observed at different incubation time (0 h or 21 h) with comet assay in vitro. Three combination-exposure ways of MW with chemicals were used: MW irradiation before chemical exposures, simultaneously exposed to MW and chemicals and MW irradiation after chemical exposures. The three chemical mutagens were mitomycin C (MMC, DNA crosslinker), bleomycin (BLM, radiometric agent), methyl methanesulfonate (MMS, alkylating agent). The exposure time of MW and chemical mutagens were 2 h and 3 h respectively.

RESULTSThe differences of comet tail length between MW group and control group were not significant when lymphocytes were incubated for 0 h or 21 h (P > 0.05). However, when lymphocytes were incubated for 21 h with 30.00 micro mol/L of MMC, the comet tail lengths of MW + MMC group, MW-MMC group and MMC + MW group were (18.00 +/- 5.96), (21.79 +/- 11.47) and (22.32 +/- 8.10) micro m respectively; while with 3.00 micro mol/L of MMC, the comet tail lengths were (8.99 +/- 3.75), (12.40 +/- 5.35) and (14.00 +/- 5.38) micro m respectively, which were significantly higher than those of corresponding MMC groups [(9.42 +/- 3.34) and (6.50 +/- 2.89) micro m, P < 0.01 or P < 0.05]. The DNA damage of MW plus BLM groups and MW plus MMS groups were not significantly different from the corresponding BLM and MMS groups (P < 0.05).

CONCLUSION2 450 MHz MW (5 mW/cm(2)) did not induce DNA damage directly, but could enhance the DNA damage effects induced by MMC. The synergistic effects of 2 450 MHz MW with BLM and MMS were not obvious.

Bleomycin ; pharmacology ; Comet Assay ; DNA ; drug effects ; genetics ; radiation effects ; DNA Damage ; Humans ; Lymphocytes ; drug effects ; metabolism ; radiation effects ; Methyl Methanesulfonate ; pharmacology ; Microwaves ; adverse effects ; Mitomycin ; pharmacology ; Mutagens ; pharmacology ; Time Factors

The association of exposure to bleomycin with the development of scleroderma-like cutaneous abnormalities has been reported. We experienced a case of scleroderma involving the hands, feet, and forearms after bleomycin chemotherapy. The present report supports the possible causal relation of bleomycin with scleroderma. Regarding the widespread use of bleomycin, this complication is thought to be under appreciated.
Bleomycin/*pharmacology ; Case Report ; Foot Dermatoses/*chemically induced/pathology/therapy ; Hand Dermatoses/*chemically induced/pathology/therapy ; Human ; Male ; Middle Age ; Scleroderma, Circumscribed/*chemically induced/pathology/therapy

Pingyangmycin (bleomycin A5 hydrochloride, PYM) is one of the anti-neoplastic agents which have been commonly used to treat venous malformations. However, the underlying mechanism by which PYM treats venous malformations remains poorly understood. It was reported that venous endothelial cells could recruit neutrophils via adhesion molecules (E-selectin, ICAM-1, ICAM-3, VCAM-1) during the acute/chronic inflammation and subsequent histological fibrosis after sclerotherapy with PYM. This study explored if the expression of E-selectin, ICAM-1, ICAM-3 and VCAM-1 in human venous malformation endothelial cells could be affected by PYM. HVMECs were cultured from human venous malformation tissue. Expressions of E-selectin, ICAM-1, ICAM-3 and VCAM-1 on HVMECs in response to PYM were analyzed by cell ELISA. The relative levels of mRNA expression in the cells were semi-quantified. The results showed that PYM up-regulated the expressions of E-selectin, ICAM-3, VCAM-1 and ICAM-1 in both time- and concentration-dependent manner. Our findings suggested that PYM could induce the expression of adhesion molecules in HVMECs, which might be a possible mechanism by which sclerotherapy by intralesional injection of PYM treats venous malformations.
Bleomycin ; analogs & derivatives ; pharmacology ; Cell Adhesion Molecules ; genetics ; metabolism ; Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; Endothelium, Vascular ; drug effects ; metabolism ; Gene Expression ; drug effects ; genetics ; Humans

OBJECTIVETo determine the effects of curcumin on bleomycin (BLM)-induced pulmonary fibrosis in rats.

METHODOne hundred and forty-four male Sprague-Dawley rats were randomized into 6 groups (24 rats in each group, model group, sham group, prednisone group (0.56 mg x kg(-1) x d(-1)), curcumin with low dose 5 mg group, curcumin with middle dose group 10 mg and curcumin with high dose group 20 mg per 100 g of body weight). Rats in all groups except in sham group were injected with BLM intratracheally. Curcumin with different doses were given by gavage one time everyday for 7, 14 and 28 days. Prednisone were given to rats in prednisone group, po, serving as the positive treatment group. On the 7th, 14th, 28th day, the lung functions (inspiratory resistance, maximal volutary ventilation, forced vital capacity, Fev 0.2/FVC, peak expiratory flow) were determinated in experimental rats, respectively, and the concentrations of hydroxyproline in lung homogenates of each rat were assayed.

RESULTAdministration of curcumin in different doses improved lung functions of BLM-induced fibrotic rats in the all experimental days; and it decreased the concentration of hydroxyproline in lung homogenates compared with those levels in model control group; and it also lessened the hyperplasia of BLM-induced pulmonary fibrosis in rats.

CONCLUSIONAdministration of curcumin can suppress BLM induced pulmonary fibrosis indicated by improved respiratory function, as well as companied with low content of hydroxyproline in lung tissue of rats.

Animals ; Bleomycin ; adverse effects ; Curcumin ; pharmacology ; Hydroxyproline ; metabolism ; Lung ; drug effects ; metabolism ; pathology ; physiopathology ; Male ; Pulmonary Fibrosis ; chemically induced ; metabolism ; pathology ; physiopathology ; Rats ; Time Factors