cyanoginosin-lr and Hepatitis

Low dose but long-term exposure of microcystin-LR (MC-LR) could induce human hepatitis and promote liver cancer according to epidemiological investigation results, but the exact mechanism has not been completely elucidated. In the present study, a chronic toxicity test of MC-LR exposure on HepG2 cells at 0.1-30 nM for 83 d was conducted under laboratory conditions. The western blot assay result revealed that MC-LR entered HepG2 cells, even at the concentration of 0.1 nM, after 83 d of exposure, but no cytotoxicity was observed in the HepG2 cells, as determined by the CCK-8 and LDH tests. However, the results of the DCF fluorescence assay showed that the intracellular ROS level in the 30 nM MC-LR-treated cells was significantly higher than that of the control cells, and 5 and 10 nM of MC-LR exposure totally increased the activity of SOD in HepG2 cells. These results indicate that MC-LR exposure at low concentration also induced excessive ROS in HepG2 cells. Additionally, long-term exposure of MC-LR at low concentration remarkably promoted the expression of NF-κB p65, COX-2, iNOS, TNF-α, IL-1β, and IL-6 in the cells, suggesting that long-term MC-LR exposure at low concentration can induce inflammatory reaction to HepG2 cells, which might account for MC-induced human hepatitis. Thus, we hypothesized that the pathogenesis of human hepatitis and hepatocarcinoma caused by MCs might be closely associated with oxidative stress and inflammation.

Topics: Bacterial Toxins; Carcinoma, Hepatocellular; Hep G2 Cells; Hepatitis; Humans; Inflammation; Interleukin-6; Liver Neoplasms; Marine Toxins; Microcystins; NF-kappa B; Oxidative Stress; Reactive Oxygen Species; Tumor Necrosis Factor-alpha

We had previously shown that microcystin-LR (MCLR) could induce lung and liver inflammation after acute exposure. The biological outcomes following prolonged exposure to MCLR, although more frequent, are still poorly understood. Thus, we aimed to verify whether repeated doses of MCLR could damage lung and liver and evaluate the dose-dependence of the results. Male Swiss mice received 10 intraperitoneal injections (i.p.) of distilled water (60 μL, CTRL) or different doses of MCLR (5 μg/kg, TOX5), 10 μg/kg (TOX10), 15 μg/kg (TOX15) and 20 μg/kg (TOX20) every other day. On the tenth injection respiratory mechanics (lung resistive and viscoelastic/inhomogeneous pressures, static elastance, and viscoelastic component of elastance) was measured. Lungs and liver were prepared for histology (morphometry and cellularity) and inflammatory mediators (KC and MIP-2) determination. All mechanical parameters and alveolar collapse were significantly higher in TOX5, 10, 15 and 20 than CTRL, but did not differ among them. Lung inflammatory cell content increased dose-dependently in all TOX groups in relation to CTRL, being TOX20 the largest. The production of KC was increased in lung and liver homogenates. MIP-2 increased in the liver of all TOX groups, but in lung homogenates it was significantly higher only in TOX20 group. All TOX mice livers showed steatosis, necrosis, inflammatory foci and a high degree of binucleated hepatocytes. In conclusion, sub-chronic exposure to MCLR damaged lung and liver in all doses, with a more important lung inflammation in TOX20 group.

Topics: Animals; Bacterial Toxins; Chemical and Drug Induced Liver Injury; Chemokine CXCL2; Chemokines; Dose-Response Relationship, Drug; Enzyme Inhibitors; Hepatitis; Injections, Intraperitoneal; Liver; Lung; Male; Marine Toxins; Mice; Microcystins; Microcystis; Organ Size; Phosphoprotein Phosphatases; Pneumonia; Random Allocation; Toxicity Tests, Subchronic

The present study aimed to determine the cytotoxicity of microcystin-LR (MC-LR) on the human hepatocellular carcinoma (HepG2) cells in order to elucidate the mechanism of apoptosis induced by MC-LR. Morphological evaluation results showed that MC-LR induced time- and concentration-dependent apoptosis in HepG2 cells. The biochemical assays revealed that MC-LR-exposure caused overproduction of reactive oxygen species (ROS), cyclooxygenase-2 activity alteration, cytochrome c release, and remarkable activation of caspase-3 and caspase-9 in HepG2 cells, indicating that MC-LR-induced apoptosis is mediated by mitochondrial pathway. Moreover, we also found that p53 and Bax might play an important role in MC-LR-induced apoptosis in HepG2 cells in which PUMA and survivin were involved. However, further studies are necessary to elucidate the possible functions of PUMA and survivin in MC-LR-induced apoptosis in HepG2 cells.

Topics: Apoptosis; Apoptosis Regulatory Proteins; Cytotoxins; Hep G2 Cells; Hepatitis; Humans; Inhibitor of Apoptosis Proteins; Liver Neoplasms; Marine Toxins; Microcystins; Proto-Oncogene Proteins; Risk Assessment; Survivin