In Vitro Cytotoxicity Testing Of Copper Sulfate And Zinc Chloride Using Different Ploidy Loach Fin Cell Lines

In this paper, the toxic effects of different concentrations of copper on diploid loach fin cell line(DIMF) and triploid loach fin cell line(TRMF),which are 70-80 generation, were studied. In the MTT measured, lethal concentrations of DIMF and TRMF were 154.3 ± 23μmol / L, 184.6 ± 14μmol / L. For zinc chloride, lethal concentrations of DIMF and TRMF were 245.2 ± 12μmol / L, 293.6 ± 23μmol / L. The lethal concentration of TRMF was significantly higher than DIMF(P <0.05). The results of antioxidase in DIMF and TRMF are as follows: superoxide dismutase(SOD) activity of DIMF gradually increased with the increasing of copper sulfate concentrations in the range of 0 ~ 200μmol/L, and reached a maximum at 200μmol/L, then gradually decreased. SOD activity of TRMF decreased with the increasing of copper sulfate concentration. Glutathione-S-transferase(GST) and glutathione peroxidase(GSH-Px) activity gradually decreased with the increasing of copper sulfate concentrations in two cell lines. After zinc chloride, these three appeared to have increased activity then decreased when the chloride concentration was 100μmol / L and the activity reached a maximum. The activity of three enzymes in TRMF were higher than DIFM. For detection the damage of copper sulfate to nuclear, micronucleus test indicated that TRMF and DIMF produced micronucleus, and the micronucleus rates gradually increased with increasing of copper sulfate concentrations. At the concentration of 400μmol/L micronucleus rates reached maximum, which the DIMF was 3.33 ‰, TRMF was 4.33‰. Ultrastructural observations: Treated with zinc chloride, zinc chloride concentration of 0~500μmol/L concentration, with the increase of concentration, the micronucleus rate increased. Concentration of 500 μmol/L, the diploid and triploid micronucleus rate achieves the maximum value, diploid was 3.33 ‰, triploid is 4.33 ‰, with the increase of the concentration after no significant change(P > 0.1).The organelles there were no significant difference between the control groups, but the vesicles of TRMF is more than DIMF. After treatment of copper sulfate, the pathological changed of DIMF and TRMF similar: the cell boundaries fuzzy, nuclear membrane structure incomplete, mitochondria disintegrated, rough endoplasmic reticulum dissolved, the ribosome drifted in the cytoplasm. Those phenomenon suggested that cell necrosis appeared.