OBJECTIVETo assess the cytotoxicity of carbon-coated iron nanoparticles (CCIN) and epirubicin-loaded CCIN on Hep-G2 cells in vitro and compare the acute toxicities of epirubicin and epirubicin-loaded CCIN in mice.

METHODSThe cytotoxicities of CCIN and epirubicin-loaded CCIN on HepG2 cells were assessed using MTT assay, and the uptake of CCIN by the tumor cells was observed by optical and electron microscopy. Different doses of epirubicin and equivalent doses of epirubicin-loaded CCIN were injected intravenously in mice to compare their acute toxicities.

RESULTSOptical and electron microscopy revealed cytoplasmic uptake of CCIN in the tumor cells without obvious destruction of the cell structural integrity. Incubation of the HepG-2 cells with different concentrations of CCIN suspension did not result in significant variation in the mean absorbance. MTT assay showed reduced cytotoxicity of epirubicin-loaded CCIN in HepG2 cells as compared with that of epirubicin alone. The cell growth inhibition rate was significantly higher with epirubicin-CCIN mixture that contained a lower proportion of CCIN. In acute toxicity experiment with mice, the median lethal dose (LD(50)) of epirubicin was 16.9 mg/kg, while that of epirubicin-CCIN mixture was 20.7 mg/kg.

CONCLUSIONCCIN uptake by HepG-2 cells does not cause obvious cytotoxicity in vitro within a certain concentration range, epirubicin-loaded CCIN has reduced cytotoxicity against HepG2 cells as compared with epirubicin, and the cytotoxicity of the mixture decreases with the increase in the CCIN content in the mixture. Epirubicin delivery in mixture with CCIN can reduce its acute toxicity in mice.

Animals ; Antibiotics, Antineoplastic ; pharmacology ; toxicity ; Carbon ; pharmacology ; toxicity ; Drug Carriers ; pharmacology ; toxicity ; Epirubicin ; pharmacology ; toxicity ; Ferric Compounds ; pharmacology ; toxicity ; Hep G2 Cells ; Humans ; Iron ; pharmacology ; toxicity ; Mice ; Nanoparticles ; toxicity ; Toxicity Tests, Acute

Although adriamycin (Doxorubicin) is one of the most effective and useful antineoplastic agents for the treatment of a variety of malignancies, its repeated administration can induce irreversible myocardial damage and resultant heart failure. Currently, no marker to detect early cardiac damage is available. The purpose of this study was to investigate whether an assessment of the acoustic properties of the myocardium could enable the earlier detection of myocardial damage after adriamycin chemotherapy. Forty Wistar rats were treated with adriamycin (2 mg/kg, i.v.) once a week for 2, 4, 6 or 8 weeks consecutively. Left ventricular ejection fraction (LVEF) was calculated using M-mode echocardiography data. The magnitude of cardiac cycle dependent variation of integrated backscatter (CVIB) of the myocardium was measured in the mid segment of the septum and in the posterior wall of the left ventricle, using a real time two dimensional integrated backscatter imaging system. LVEF was significantly lower in the adriamycin-treated 8-week group than in the controls (75 +/- 9 vs 57 +/- 8%, p < 0.05). Myocyte damage was only seen in the 8-week adriamycin-treated group. However, no significant changes of CVIB were observed between baseline or during follow-up in the ADR or control group. In conclusion, serial assessment of the acoustic properties of the myocardium may not be an optimal tool for the early detection of myocardial damage after doxorubicin chemotherapy in a rat model.
Animals ; Antibiotics, Antineoplastic/*toxicity ; Cardiomyopathies/*chemically induced/*ultrasonography ; Disease Models, Animal ; Doxorubicin/*toxicity ; *Echocardiography ; Male ; Rats ; Rats, Wistar ; Research Support, Non-U.S. Gov't

OBJECTIVETo observe the ultrastructures of rat alveolar type II cells and change of composition of phospholipid(PL) and content of protein in pulmonary surfactant(PS), to investigate the relation between change in composition of PL and activity of alveolar type II cells.

METHODSThe rats lung injury models were made by intratracheally instilling bleomycin(BLM) (4 mg/ml, 5 mg/kg). 28 rats were divided into four groups: 3-day group, 7-day group, 14-day group and 28-day group. Preparations of each group were stained histochemically and examined by electron microscope, content of PL in BALF, composition of PL and content of protein of each group were determined respectively.

RESULTS(1) Rats lungs in experimental groups were found that PS lost continuously, appeared homogenous and chorionic, dropped in the pulmonary alveolies. 3-day group was more apparent. Ruthenium red attaching on pulmonary surfactant was thicker in 3-day group, and the colour deeper, no difference in 7-day group and 14-day group, thinner in 28-day group. Content of PL in PS of BALF was increasing. Content of phosphatidylglycerol(PG) increased in 3-day group, decreased in 7-day, 14-day and 28-day group. The change of content of phosphatidylinositol(PI) was reversed. (2) Alveolar type II cells degenerated, necrotized, even disintegrated in 3-day group and 7-day group. 3-day group was more apparent. Proliferations of alveolar type II cells were found in each group, 7-day group was more apparent. We found that type II cells transformed to type I cells in 14-day group, extended and attached on bare basement membrane. Content of protein in PS was the highest in 3-day group, almost equal to the content of the control group in 28-day group.

CONCLUSIONMorphologic change and alternation of quality and quantity of PS after bleomycin-induced pulmonary injures specifically reflect the activity of alveolar type II cells. Measuring content of PL in BALF is one of simple and feasible method judging activity of alveolar type II cells when lungs of the rats are injured early by bleomycin.

Animals ; Antibiotics, Antineoplastic ; toxicity ; Bleomycin ; toxicity ; Bronchoalveolar Lavage Fluid ; chemistry ; Lung ; drug effects ; pathology ; ultrastructure ; Pulmonary Alveoli ; chemistry ; Pulmonary Surfactants ; analysis ; Rats

OBJECTIVEThe pathogenesis of minimal change nephrotic syndrome (MCNS) remains unclear. This study aimed to investigate the expression of interleukin-6 (IL-6) in rats with doxorubicin-induced nephropathy and its possible roles in the pathogenesis of MCNS.

METHODSEighty-three male Wistar rats were randomly assigned into a control group (n=32) and a nephropathy group (n=51). Nephropathy was induced by a single tail vein injection of doxorubicin (5 mg/kg). The control group was injected with normal saline. Twenty-four-hour urinary protein excretion was measured 7, 14, 28 and 42 days after doxorubicin injection. IL-6 expression in urine and renal tissues was determined using ELISA 7, 14, 28 and 42 days after doxorubicin injection.

RESULTSThe urinary protein excretion increased significantly in the nephropathy group 7, 14, 28 and 42 days after doxorubicin injection compared with that in the control group (P<0.01). IL-6 expression in urine and renal tissues increased significantly 7, 14, 28 and 42 days after doxorubicin injection compared with that in the control group (P<0.01). IL-6 expression in urine and renal tissues was positively correlated with 24-hour urinary protein excretion in the nephropathy group (r=0.794, P<0.01; r= 0.870, P<0.01). IL-6 expression in urine was positively correlated with that in renal tissues (r=0.739, P<0.01).

CONCLUSIONSIL-6 expression in the urine and renal tissues is increased in MCNS rats. IL-6 might play an important role in the pathogenesis of MCNS.

Animals ; Antibiotics, Antineoplastic ; toxicity ; Disease Models, Animal ; Doxorubicin ; toxicity ; Interleukin-6 ; analysis ; Kidney ; chemistry ; Male ; Nephrosis, Lipoid ; chemically induced ; immunology ; Rats ; Rats, Wistar

OBJECTIVETo study the effects and mechanism of Buzhong Yiqi decoction on adriamycin-induced acute myocardial injury in rats.

METHOD50 rats were randomly divided to five groups: control group, heart failure group, low dose Buzhong Yiqi decoction, high dose Buzhong Yiqi decoction and captopril group. Adriamycin was injected into the latter four groups to built a model of heart failure. Then, the effects of different doses of Buzhong Yiqi decoction on hemodynamics, cardiac tissue histological changes, antioxidant capacity and apoptosis of the damaged hearts were studied.

RESULTAdriamycin led to myocardial fiber swelling and fracture, Buzhong Yiqi decoction could reduce myocardial lesions. Buzhong Yiqi decoction could also improve heart antioxidant capacity and inhibit adriamycin-induced cardiomyocyte apoptosis.

CONCLUSIONBuzhong Yiqi decoction could significantly ease adriamycin induced heart failure in rats, and the mechanism is related to anti-oxidation and inhibiting apoptosis.

Animals ; Antibiotics, Antineoplastic ; toxicity ; Doxorubicin ; toxicity ; Drugs, Chinese Herbal ; pharmacology ; Heart Failure ; chemically induced ; prevention & control ; Male ; Nitric Oxide ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood ; Ventricular Function, Left ; drug effects

OBJECTIVETo determine the in vitro possible clastogenic and cytotoxic activities of Ulva rigida crude extracts (URE), and identify their antigenotoxic and protective effects on chemotherapeutic agent mitomycine-C (MMC).

METHODSAnti-clastogenic and anti-genotoxic activities of Ulva rigida crude extracts (URE) were studied using chromosome aberration (CA), sister chromatid exchange (SCE), and micronuclei (MN) tests in human lymphocytes cultured in vitro.

RESULTSThe chromosome aberration, sister chromatid exchange or micronuclei tests showed that URE at concentrations of 10, 20, and 40 microg/mL had no clastogenic activity in human lymphocyte cell culture. Three doses of URE significantly decreased the number of chromosomal aberrations and the frequencies of SCE and MN when compared with the culture treated with MMC (P < 0.0001).

CONCLUSIONAlthough URE itself is not a clastogenic or cytotoxic substance, it possesses strong antigenotoxic, anti-clastogenic, and protective effects on MMC in vitro.

Antibiotics, Antineoplastic ; pharmacology ; Antimutagenic Agents ; pharmacology ; Cells, Cultured ; Chlorophyta ; Chromosome Aberrations ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Lymphocytes ; drug effects ; metabolism ; Micronucleus Tests ; Mitomycins ; pharmacology ; Mutagens ; toxicity ; Plant Extracts ; chemistry ; pharmacology ; Sister Chromatid Exchange ; drug effects

PURPOSE: Matrix metalloproteinases (MMPs) have been implicated in the pathogenesis of pulmonary fibrosis. To understand the role of MMP-2 and MMP-9 in pulmonary fibrosis, we evaluated the sequential dynamic change and different cellular sources of the 2 MMPs along the time course and their differential expression in the bronchoalveolar lavage (BAL) fluid and in the lung parenchyma of the bleomycin-induced pulmonary fibrosis models in rats. MATERIALS AND METHODS: The level of MMPs in BAL fluid of 54 bleomycin-treated rats was assessed by zymography from 1 to 28 days after intratracheal bleomycin instillation. The level of MMPs in lung parenchyma was evaluated by immunohistochemistry. RESULTS: MMP-2 and MMP-9 were markedly increased in both the BAL fluid and in the lung parenchyma of the bleomycin-treated rats, especially in the early phase with the peak on the 4th day. The levels of both MMPs in the BAL fluid correlated generally well to those in lung parenchyma, although the level of MMP-9 in BAL fluid was higher than MMP-2. In the lung parenchyma, the 2 MMPs, in early stage, were predominantly expressed in the inflammatory cells. In late stage, type II pneumocytes and alveolar epithelial cells at the periphery of the fibrotic foci retained MMP expression, which was more prominent in the cells showing features of cellular injury and/or repair. CONCLUSION: In bleomycin-induced pulmonary fibrosis, MMP-2 and MMP-9 may play important roles, especially in the early phase. In the late stage, the MMP-2 and MMP-9 may play a role in the process of repair.
Animals ; Antibiotics, Antineoplastic/toxicity ; Bleomycin/toxicity ; Bronchioles/*enzymology/pathology ; Bronchoalveolar Lavage Fluid/cytology/immunology ; Disease Models, Animal ; Enzyme Activation ; Gelatin ; Immunohistochemistry ; Male ; Matrix Metalloproteinase 2/*metabolism ; Matrix Metalloproteinase 9/*metabolism ; Neutrophils/pathology ; Pulmonary Fibrosis/chemically induced/*metabolism/*pathology ; Rats ; Rats, Sprague-Dawley

AIMMultidrug resistance ( MDR) as a major obstacle to successful clinical cancer chemotherapy, searching a novel effective antiresistant drug would be necessary.

METHODSA novel doxorubicin anti-resistant stealth liposomes (DARSLs) was prepared by co-encapsulating doxorubicin (DOX) and verapamil (VER) into stealth liposomes with ammonium sulfate gradient remote loading approach. In vitro cytotoxity of various DOX formulations and in vivo toxicity of DARSLs were evaluated using DOX-resistant rat prostate cancer cell line (MLLB2), human uterus sarcoma cell line (MES-SA/DX5) and normal SD rats, separately.

RESULTSThe DARSLs liposome suspensions mainly consisted of homogeneous large unilamellar vesicles (LUV) with average particle size of (118.1 +/- 22.3) nm. Encapsulation efficiencies of DOX and VER in DARSLs were more than 90% and about 70%, respectively, when the ratio of DOX/VER/Lipid was 1: 0.11 :10 (w/w/w). In vitro cytotoxicity tests of the DARSLs using rat prostate cancer cell line (MLLB2) and human uterus sarcoma cell line (MES-SA/DX5) showed that 5 micromol x L(-1) VER significantly reversed DOX-resistance of these 2 cell lines and DARSLs was the most effective on inhibition of DOX-resistant cell growth. Besides, compared to FDFV, much slower DOX distribution (confocal microscopy) to nuclei and cytoplasm in MLLB2 cells for DARSLs suggested that it might possess distinct mechanism of cytotoxicity. Systemic and cardiac toxicity evaluations in normal SD rats suggested that liposomal encapsulation could significantly improve the severe cardiotoxicity arising from simultanous administration of DOX and VER.

CONCLUSIONDARSLs is a novel anticancer liposome formulation with lower cardiotoxicity, effective drug-resistance reversal and intravenous injection.

Animals ; Antibiotics, Antineoplastic ; administration & dosage ; pharmacology ; toxicity ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Doxorubicin ; administration & dosage ; pharmacology ; toxicity ; Drug Carriers ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Female ; Heart Rate ; drug effects ; Humans ; Liposomes ; Male ; Myocytes, Cardiac ; drug effects ; ultrastructure ; Prostatic Neoplasms ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sarcoma ; pathology ; Uterine Neoplasms ; pathology

OBJECTIVETo investigate the trend and potential pathogenic role of nuclear factor (NF)-kappaB P(65)/Rel-A mRNA and angiotensin-II (AngII) receptor type 1 (AT(1)) proteins expression, and the relativity between them in early stage of renal tubulointerstitial lesions in young rats with adriamycin nephrosis and the interfering effects of treatment with angiotensin converting enzyme inhibitor (ACEI) benazepril and ACEI combined with AngII type 1 receptor antagonist (AT(1)RA) Losartan.

METHODSMale young Wistar rats with adriamycin nephrosis were used as experimental models. At different time points (weeks 1, 2, and 3 in early nephritic phase, the urinary protein and blood biochemical parameters were measured, and P(65)/Rel-A mRNA was detected; AT(1) protein expression was determined by in situ hybridization and immunohistochemical methods. The relativity between them was evaluated.

RESULTSIn the early phase of tubulointerstitial lesions, following adriamycin injection and proteinuria aggravated progressively, at week 3, the proteinuria level had reached heavy proteinuria (123.2 +/- 7.7 mg/24 h). The serum parameters reflecting renal function were elevated. The inflammatory cells infiltrated into renal tissues, especially in tubulointerstitial regions, were increased markedly. Swelling of tubular epithelial cells, broadened tubulointerstitial areas, and protein casts in tubule were observed. In situ hybridization and immunochemical staining showed that AT(1) protein was expressed in tubular epithelial cell cytoplasm and on nuclear membranes (AT(1): 1st week 19.8 +/- 1.1%, 2nd week 25.0 +/- 2.6%, 3rd week 37.1 +/- 1.0% (control: 10.3 +/- 0.8%, 10.4 +/- 1.6%, 10.2 +/- 1.5%); and P(65)/Rel-A mRNA expression in the same locations was upregulated. P(65)/Rel-A translocation from cytoplasm into nucleus increased markedly simultaneously. The positive signal of hybridization dominated in cytoplasm gradually became dominant in the nuclei as the pathological changes progressed. The semiquantitative expression of P(65)/Rel-A was 24.0 +/- 3.3% at week 1, 34.2 +/- 2.4% at week 2, 39.9 +/- 6.4% at week 3, while the values of controls were 8.5 +/- 0.4%, 8.7 +/- 1.0%, and 8.4 +/- 0.9%, respectively. There was a positive correlation between AT(1) and P(65)/Rel-A expression in localization and time phase (r = 0.857, P < 0.01). However, the tendency of those factor's expression was all decreased in each treated group, the semiquantitative results were AT(1): 14.6 +/- 2.1%, 13.7 +/- 2.3%, 11.4 +/- 1.1%; P(65)/Rel-A: 18.5 +/- 3.4%, 22.8 +/- 1.6%, 26.7 +/- 4.9% at 1, 2, 3 weeks in ACEI treated group; AT(1): 12.4 +/- 1.5%, 11.1 +/- 1.0%, 10.3 +/- 0.8%; P(65)/Rel-A: 17.9 +/- 5.0%, 21.3 +/- 6.0%, 22.5 +/- 2.5% in AT(1)RA (Losartan) group, respectively. The significant difference were observed between all groups in different time points (P < 0.05).

CONCLUSIONSThe present study suggested that NF-kappaB (P(65)/Rel-A) mRNA expression and its activity was enhanced significantly that synchronized with aggravating injures in tubulointerstitial lesions initial period induced by proteinuria-loading in nephrotic young rats. This tendency was related with AngII and its receptors system that may accelerate lesions progressing in many renal diseases.

Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Antibiotics, Antineoplastic ; toxicity ; Benzazepines ; pharmacology ; Biopsy ; Disease Models, Animal ; Doxorubicin ; toxicity ; Gene Expression Regulation ; drug effects ; Immunohistochemistry ; In Situ Hybridization ; Kidney ; drug effects ; metabolism ; pathology ; Losartan ; pharmacology ; Male ; NF-kappa B ; genetics ; metabolism ; Nephrosis ; chemically induced ; drug therapy ; metabolism ; Rats ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Transcription Factor RelA

This study is to observe the protection of gross saponins of Tribulus terrestris (GSTT) on cardiocytes impaired by adriamycin (ADR) and approach its mechanism of action. Cardiocytes of neonate rat were cultivated for 72 hours and divided into normal control group, model (ADR 2 mg x L(-1)) group, and GSTT (100, 30, and 10 mg x L(-1)) groups. MTT colorimetric method was deployed to detect cardiocyte survival rate, activities of CK, LDH, AST, SOD, MDA and NO were detected, and apoptosis was detected with flow cytometry. Effect of GSTT on caspase-3 was detected with Western blotting. Compared with control group, contents of CK, LDH, AST, MDA and NO were increased, and activity of SOD was reduced (P < 0.05, P < 0.01, P < 0.001) by ADR. Numbers of survival cells were increased (P < 0.05, P < 0.001), contents of CK, LDH, AST, MDA and NO were decreased, and activity of SOD was increased (P < 0.05, P < 0.01, P < 0.001) by GSTT (100 and 30 mg x L(-1)). Apoptosis of cardiocytes and concentration of caspase-3 can be reduced by GSTT (100 and 30 mg x L(-1)). GSTT can protect cardiocytes impaired by ADR, which are possible involved with its effect of resisting oxygen free radical.
Animals ; Antibiotics, Antineoplastic ; toxicity ; Apoptosis ; drug effects ; Aspartate Aminotransferases ; metabolism ; Caspase 3 ; metabolism ; Cell Survival ; drug effects ; Cells, Cultured ; Creatine Kinase ; metabolism ; Doxorubicin ; toxicity ; L-Lactate Dehydrogenase ; metabolism ; Malondialdehyde ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Nitric Oxide ; metabolism ; Rats ; Rats, Wistar ; Saponins ; isolation & purification ; pharmacology ; Superoxide Dismutase ; metabolism ; Tribulus ; chemistry