ABSTRACT
OBJECTIVE: To explore the toxicity of 1,2-dichloroethane( 1,2-DCE) and its metabolites on human astrocytes( HAs). METHODS: Different doses of 1,2-DCE( 5. 00,10. 00,25. 00,50. 00 and 100. 00 mmol/L),2-chlorohydrins( 5. 00,25. 00,50. 00,100. 00 and 200. 00 mmol/L),2-chloroacetaldehyde( 1. 00,5. 00,10. 00,20. 00 and 50. 00 mmol / L) and chloroacetic acid( 0. 01,0. 05,0. 10,0. 50 and 1. 00 mmol / L) were used for treating HAs in vitro during their logarithmic phase. After 24 hours of culture,the morphology of HAs was observed by fluorescent inverted phase contrast microscope. The survival rate and the inhibition ratio of HAs were detected by CCK-8 colorimetry to estimate the50% inhibiting concentration in 24 hours( 24 h-IC50). The apoptosis of HAs was tested by double-labeling and flow cytometry using Annexin Ⅴ-fluorescein isothiocyanate and propidium iodide. RESULTS: The morphology of HAs changed in varying degrees after 24 hours exposure to 1,2-DCE,2-chlorohydrins,2-chloroacetaldehyde and chloroacetic acid. The changes included smaller size of cells,pseudopodia tapering,increased intracellular particles and suspension of circular cells and decreased transparency of cells. With the increasing does of 1,2-DCE,2-chlorohydrins,2-chloroacetaldehyde and chloroacetic acid exposure,the survival rates of HAs decreased( P < 0. 01),while its inhibition ratios increased( P <0. 01). They all showed dose-effect relationship. 24 h-IC50 of the above 4 chemicals were 56. 25,235. 00,26. 43 and1. 38 mmol / L,respectively. The 1,2-DCE,2-chlorohydrins and chloroacetic acid could induce the apoptosis of HAs and the apoptosis rate of HAs was positively correlated with the 3 kinds of chemicals( P < 0. 01). CONCLUSION: 1,2-DCE and its metabolites 2-chloroacetaldehyde,2-chlorohydrins and chloroacetic acid can lead to toxic damage and induce the apoptosis of HAs. Chloroacetic acid has the strongest toxicity among the metabolites.
ABSTRACT
It was reported that a complete antifertility action of alpha-chlorohydrin in combination with glycidol in male rats after a daily oral dose of 5mg/kg and 75mg/kg respectively for two days. Simultaneously some ultrastructural changes including mitochondrial swelling, atrophy of Golgi complex, decreasing in pinocytotic vesicles and stereocilia were found in principal cells of epididymis within 3 days after medication.In this experiment the animals received 3, 6 and 9 courses of treatment respectively, each course included two days of administration with a daily dose mentioned above and eight days of cessation of medication.The specimens of initial segment of epididymis, liver and kidney were taken at the end of 3 rd, 6 th and 9th courses of treatment for electron microscopy.The principal cells of initial segment of epididymis revealed a normal feature at the end of 3 rd and 9 th course of treatment. But at the end of 6 th course of treatment, the atrophy of Golgi complex and the decrease in stereocilia and pinocytotic vesicles were found in some principal cells.Any ultrastructural changs of sperms in the initial segment were not found by electron microscope.There was also mitochondrial changes with striking increase in the density of the matrix and reduction of size in some liver cells at the end of 6 th course of treatment, but not at the end of 3 rd or 9 th course of treatment. The epithelial cells of nephron in all the specimens revealed a normal ultrastructural feature.The results suggest that the ultrastructural changes of the principal cells and the liver cells induced by the mixture of the drugs could be recovered at the end of treatments.