The diethyl sulfate (DES) mutagenesis was chosen for the mutagenic treatment to Phellinus igniarius, and the relationship of mutagenesis time and death rate was investigated with 0.5% DES. The differences of mycelial growth speed, liquid fermentation mycelia biomass, morphology and pigment classes of secondary metabolites production speed and antioxidant activities of metabolite products were discussed. The study displayed that DES mutagenesis could change mycelial morphology without obvious effect on mycelium growth, and the DES mutagenesis improved antioxidant activities of the active ingredients of P. igniarius and had more antioxidant activity of hypoxia/sugar PC12 nerve cells than that of P. igniarius.
Basidiomycota ; drug effects ; genetics ; growth & development ; metabolism ; Mutagenesis ; Mutagens ; pharmacology ; Mycelium ; drug effects ; genetics ; growth & development ; metabolism ; Pigments, Biological ; analysis ; metabolism ; Secondary Metabolism ; drug effects ; Sulfuric Acid Esters ; pharmacology

OBJECTIVETo study the effect of Morinda officinalis (MO) extract on cytoxan (CTX) -impaired spermatogenesis of adult male SD rats.

METHODSWe randomly divided 56 adult male SD rats into seven groups of equal number: blank control, CTX model, CTX + NS, CTX + 10 g/kg MO, CTX + 20 g/kg MO, CTX + 30 g/kg MO, and CTX + 40 g/kg MO. We made the models of impaired spermatogenesis in the SD rats by intraperitoneal injection of CTX and treated the animal models by intragastric administration of MO at the concentrations of 10, 20, 30, and 40 g per kg per d, respectively. After two weeks of medication, we observed the changes in the body weight, testicular and epididymal indexes, and microstructure of the testis tissue, measured the mean seminiferous tubule diameter (MSTD) , and obtained testicular biopsy scores (TBS) in different groups, followed by comparative analyses.

RESULTSAfter treatment, the CTX + NS group showed no remarkable differences in the body weight ([234.83 ± 28.77] g) and epididymal index (2.71 ± 0.34) from those of the four CTX + MO groups, but exhibited a significantly lower testicular index ([12.15 ± 1.04] g) than those in the CTX + 20 g/kg MO ([13.71 ± 0.97] g), CTX + 30 g/kg MO, ([13.30 ± 0.29] g), and CTX + 40 g/kg MO group ([13.48 ± 0.51] g) (P < 0.05). Light microscopy revealed obvious pathological changes of the testis tissue in the CTX + NS group and significantly ameliorated structures of the seminiferous tubules in the CTX + MO 10, 20, 30, and 40 g/kg groups, with the MSTD of (204.78 ± 11.03), (216.55 ± 10.93), (218.03 ± 11.23), and (218.59 ± 14.06) μm, respectively, and the TBS of 9.03 ± 0.39, 9.69 ± 0.26, 9.83 ± 0.18, and 9.89 ± 0.11, respectively, all significantly higher than (189.74 ± 8.55) μm and 5.95 ± 1.21 in the CTX + NS group (P < 0.05). The efficacy of MO extract was increased in a concentration-dependent manner.

CONCLUSIONMorinda officinalis extract can repair cytoxan-induced damage to rat spermatogenesis, with may achieve the best effect at the concentrations of 30 and 40 g per kg per d.

Animals ; Body Weight ; drug effects ; Cyclophosphamide ; toxicity ; Epididymis ; drug effects ; Male ; Morinda ; chemistry ; Mutagens ; toxicity ; Plant Extracts ; administration & dosage ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; drug effects ; pathology ; Spermatogenesis ; drug effects ; Testis ; drug effects ; ultrastructure

This study is designed to evaluate antioxidant and antigenotoxic activities of corn tassel extracts (CTTs). The major bioactive components of CTTs include flavonoid, saponin and polysaccharide. The antioxidant properties of the three bioactive components of CTTs were investigated by Ferric Reducing Antioxidant Property (FRAP) and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assays. The activities of the extracts were determined by assessing the inhibition of mutagenicity of the direct-acting mutagen fenaminosulf, sodium azide, and indirect-acting mutagen 2-aminofluorene using the Ames test (strains TA98 and TA100). The results showed that the extraction rates of flavonoid, saponin, and polysaccharide from the dried corn tassels were 1.67%, 2.41% and 4.76% respectively. DPPH and FRAP assay strongly demonstrated that CTTs had antioxidant properties. CTTs at doses of 625, 1250 and 2500 μg per plate reduced 2-aminofluorene mutagenicity by 12.52%, 28.76% and 36.49% in Salmonella typhimurium TA98 strain assay respectively and by 10.98%, 25.27% and 37.83%, at the same doses in Salmonella typhimurium TA100 assay system, respectively. 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay showed that the different concentrations of CTTs inhibited the proliferation of MGC80-3 cells in a dose-dependent manner (P<0.01). It is concluded that these integrated approaches to antioxidant and antigenotoxicity assessment may be useful to study corn tassel as a natural herbal material.
Antimutagenic Agents ; pharmacology ; Antioxidants ; pharmacology ; Biphenyl Compounds ; antagonists & inhibitors ; metabolism ; Cell Line, Tumor ; Cell Survival ; drug effects ; Flavonoids ; pharmacology ; Fluorenes ; pharmacology ; Free Radical Scavengers ; pharmacology ; Humans ; Inflorescence ; chemistry ; Mutagens ; pharmacology ; Picrates ; antagonists & inhibitors ; metabolism ; Plant Extracts ; pharmacology ; Polysaccharides ; pharmacology ; Salmonella typhimurium ; drug effects ; genetics ; Saponins ; pharmacology ; Zea mays ; chemistry

OBJECTIVETo investigate the protective effect of epigallocatechin gallate (EGCG) on mouse sperm in vivo.

METHODSA total of 64 six-week-old male Kuming mice were randomly divided into eight groups of equal number to be treated with normal saline (negative control), Cyclophosphamide (CP) at 30 mg/kg (positive control), and CP followed by EGCG (experimental) at 20, 40, and 80 mg/kg, respectively, given every other day for 10 days. At 4 and 5 weeks after treatment, the bilateral testes of the mice were harvested for examination of sperm abnormality.

RESULTSEGCG did not increase the rate of CP-induced sperm abnormality in the mice, but reduced it instead with the prolonged time of treatment.

CONCLUSIONEGCG protects mouse sperm in vivo.

Animals ; Catechin ; analogs & derivatives ; pharmacology ; Cyclophosphamide ; toxicity ; Male ; Mice ; Mutagens ; toxicity ; Random Allocation ; Spermatozoa ; drug effects ; Time Factors

OBJECTIVEThe purpose of this study was to investigate the effects of exogenous epidermal growth factor (EGF) on the growth inhibition of renal proximal tubular epithelial cell induced by AA-I.

METHODCultured human renal proximal tubular epithelial cell line HK-2 was used as the subject. The changes of the survived HK-2 cells were observed and compared among control, AA-I stimulation, pre-EGF, together-EGF and post-EGF groups. In the study, cellular morphologic assessments were performed with a phase-contrast inverted microscope. Cell counting after stained with 0.04% trypan blue was adopted to analyze cell proliferation. Cell cycle was assessed by flow cytometry.

RESULTNumber of the survived cells stimulated by AA-I for 12, 24, 48 hours decreased gradually in a dose and time dependent manner. At 24, 48 hours, the survived cells showed a significant disturbance in the cell cycle procedure, which was characterized as decreased percentage of cells in G0/G1 phase, significant increased percentage of cells in G2/M phases. Exogenous EGF (20 microg L(-1)) could significantly promote the proliferation of HK-2 cells, which shown a increased cell number, accompanied down-regulated cells in G0/G1 phase, increased cells in S and G2/M phase. Compared with AA-I groups, it failed to improve the inhibitory effect on cell proliferation and abnormal cell cycle procedure by AA-I, no matter EGF was added in before, at same time or after AA-I stimulation.

CONCLUSIONAA-I (10 g L(-1)) has remarkable growth inhibition effects on survived RTEC, and can induce a blockage of G2/M phase in the cell cycle procedure. Exogenous EGF (20 microg L(-1)) promote the proliferation in normal cultured HK-2 cell. EGF treatment could not improve the proliferation inhibitory effect induced by AA-I, no matter adding EGF to cultures before, together with AA-I or after AA-I stimulation.

Aristolochic Acids ; pharmacology ; Cell Cycle ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Epidermal Growth Factor ; pharmacology ; Epithelial Cells ; drug effects ; Flow Cytometry ; Humans ; Kidney Tubules, Proximal ; cytology ; drug effects ; Mutagens ; pharmacology

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

OBJECTIVETo study the modulatory effect of distillate of Ocimum sanctum (traditionally known as Tulsi) leaf extract (DTLE) on genotoxicants.

METHODSIn the present investigation, we studied the antigenotoxic and anticlastogenic effect of distillate of Tulsi leaf extract on (i) human polymorphonuclear leukocytes by evaluating the DNA strand break without metabolic activation against mitomycin C (MMC) and hexavalent chromium (Cr+6) and (ii) human peripheral lymphocytes (in vitro) with or without metabolic activation against mitomycin C (MMC), hexavalent chromium (Cr+6) and B[a]P by evaluating chromosomal aberration (CA) and micronucleus assay (MN). Three different doses of DTLE, 50 microL/mL, 100 microL/mL, and 200 microL/mL were selected on the basis of cytotoxicity assay and used for studying DNA strand break, chromosomal aberration and micronucleus emergence. The following positive controls were used for inducing genotoxicity and clastogenicity: MMC (0.29 micromol/L) for DNA strand break, chromosomal aberration and 0.51 micromol/L for micronucleus assay; Potassium dichromate (Cr+6) 600 micromol/L for DNA strand break and 5 micromol/L for chromosomal aberration and micronucleus assay; Benzo[a]pyrene (30 micromol/L) for chromosomal aberration and 40 micromol/L for micronucleus assay. The active ingredients present in the distillate of Tulsi leaf extract were identified by HPLC and LC-MS.

RESULTSMitomycin C (MMC) and hexavalent chromium (Cr+6) induced statistically significant DNA strand break of respectively 69% and 71% (P<0.001) as revealed by fluorometric analysis of DNA unwinding. Furthermore, the damage could be protected with DTLE (50 microL/mL, 100 microL/mL, and 200 microL/mL) on simultaneous treatment. Chromosomal aberration and micronucleus formation induced by MMC, Cr+6 and B[a]P were significantly protected (P<0.001) by DTLE with and without metabolic activation.

CONCLUSIONDistillate of Tulsi leaf extract possesses antioxidants contributed mainly by eugenol, luteolin and apigenin as identified by LC-MS. These active ingredients may have the protective effect against genotoxicants.

Adult ; Benzopyrenes ; toxicity ; Cell Survival ; drug effects ; Chromium ; toxicity ; Chromosome Aberrations ; drug effects ; DNA ; metabolism ; DNA Damage ; drug effects ; Humans ; Lymphocytes ; drug effects ; Mass Spectrometry ; Mitomycin ; toxicity ; Mutagens ; toxicity ; Ocimum ; chemistry ; Plant Extracts ; pharmacology ; Plant Leaves ; chemistry

Turmeric has long been used as a spice and food colouring agent in Asia. In the present investigation, the antimutagenic potential of curcumin was evaluated in Allium cepa root meristem cells. So far there is no report on the biological properties of curcumin in plant test systems. The root tip cells were treated with sodium azide at 200 and 300 microg/ml for 3 h and curcumin was given at 5, 10 and 20 microg/ml for 16 h, prior to sodium azide treatment. The tips were squashed after colchicine treatment and the cells were analyzed for chromosome aberration and mitotic index. Curcumin induces chromosomal aberration in Allium cepa root tip cells in an insignificant manner, when compared with untreated control. Sodium azide alone induces chromosomal aberrations significantly with increasing concentrations. The total number of aberrations was significantly reduced in root tip cells pretreated with curcumin. The study reveals that curcumin has antimutagenic potential against sodium azide induced chromosomal aberrations in Allium cepa root meristem cells. In addition, it showed mild cytotoxicity by reducing the percentage of mitotic index in all curcumin treated groups, but the mechanism of action remains unknown. The antimutagenic potential of curcumin is effective at 5 microg/ml in Allium cepa root meristem cells.
Antimutagenic Agents ; pharmacology ; Chromosome Aberrations ; drug effects ; Curcumin ; pharmacology ; Meristem ; drug effects ; genetics ; Mutagens ; toxicity ; Onions ; drug effects ; genetics ; Sodium Azide ; toxicity

OBJECTIVETo study the anticlastogenic effect of redistilled cow's urine distillate (RCUD) in human peripheral lymphocytes (HLC) challenged with manganese dioxide and hexavalent chromium.

METHODSThe anticlastogenic activity of redistilled cow's urine distillate was studied in human polymorphonuclear leukocytes (HPNLs) and human peripheral lymphocytes in vitro challenged with manganese dioxide and hexavalent chromium as established genotoxicants and clastogens which could cause induction of DNA strand break, chromosomal aberration and micronucleus. Three different levels of RCUD: 1 microL/mL, 50 microL/mL and 100 microL/mL, were used in the study.

RESULTSManganese dioxide and hexavalent chromium caused statistically significant DNA strand break, chromosomal aberration and micronucleus formation, which could be protected by redistilled cow's urine distillate.

CONCLUSIONThe redistilled cow's urine distillate posseses strong antigenotoxic and anticlastogenic properties against HPNLs and HLC treated with Cr+6 and MnO2. This property is mainly due to the antioxidants present in RCUD.

Animals ; Antimutagenic Agents ; pharmacology ; Antioxidants ; pharmacology ; Cattle ; urine ; Cells, Cultured ; Chromium ; antagonists & inhibitors ; toxicity ; DNA Damage ; Humans ; Hydrogen-Ion Concentration ; Lymphocytes ; drug effects ; Manganese Compounds ; antagonists & inhibitors ; Mutagenicity Tests ; Mutagens ; toxicity ; Oxides ; antagonists & inhibitors ; toxicity ; Urine ; chemistry

OBJECTIVETo study the effect of hydroquinone on apoptosis of bone marrow mononuclear cells, and to evaluate the toxic effect of benzene on stem cells.

METHODSCell morphology was observed by HT fluorescent stain method, and DNA fragments were analyzed by agarose gel electrophoresis. Anti-Annexin V FITC plus PI staining for apoptotic and necrotic rate was examined by flow cytometer.

RESULTSAfter adding different concentrations of hydroquinone to the cells for 6 h culture, the fluorescent intensity of nucleus increased, the color of nucleus became deep and inhomogeneous, and the chromatin was condensed and distributed around the neucleus. DNA ladder was detected in all samples. Cell apoptotic rate in different concentration of hydroquinone groups was significantly higher than that in blank control group (P < 0.05). With the increase of the concentration of hydroquinone, the apoptotic and necrotic rate also increased. The optimal concentration of hydroquinone was 50 micro mol/L. When it was >or= 75 micro mol/L, the necrotic rate increased significantly. Hydroquinone-induced apoptosis was associated with culture time at the concentration of 50 micro mol/L, and the peak apoptotic time was 10 h, then the apoptotic rate decreased and necrotic rate increased.

CONCLUSIONHydroquinone can induce apoptosis of bone marrow mononuclear cells in vitro with dose-effect and time-effect relationship.

Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Humans ; Hydroquinones ; pharmacology ; Leukocytes, Mononuclear ; cytology ; drug effects ; Mutagens ; pharmacology