Sodium monochloroacetate causes cytotoxic effects,an increased lactate and pyruvate level and induces ultra structural and cytoskeletal alterations in cultured kidney and liver epithelial cells

Abstract

1 Monochloroacetic acid (MCAA) and its sodium salt, sodium monochloroacetate (SMCA) are widely used in chemical industries as intermediates in the synthesis of carboxymethylcellulose, phenoxyacetic acid, thio-glycolic acid, glycine, indigoid dyes and others. More-over, MCAA has been found as a by-product of the chlorination disinfection of drinking water and as an environmental contaminant of the atmosphere from the photodechlorination reactions of chlorinated hydrocarbons. Little is known about the mode of action of both compounds on the cellular level. From cases of accidental poisoning of man it is known that MCAA accumulates in liver and kidney.

2 In this study, the cytotoxicity of SMCA on cultured liver (Chang liver cells) and kidney epithelial cells of the proximal tubule (Opossum kidney cells) was investigated and its effect on metabolism, ultrastruc-ture and organization of cytoskeleton was examined.

3 Independent from the growth state of the cells (proliferating or quiescent), the results clearly show that SMCA causes a dose-dependent decrease in cell viability after an exposure period of 24 h. In all experiments, proliferating cells were more sensitive than quiescent and confluent cells. Liver cells were less sensitive against SMCA treatment than kidney epithe-lial cells. In contrast to liver cells, kidney cells exhibited a dose-dependent decrease in cell volume. The decrease in cell viability was accompanied by an increase of lactate and pyruvate concentrations released into the culture medium. In the case of Opossum kidney cells, lactate and pyruvate levels increased 5-6-fold, whereas in the case of Chang liver cells the increase was approximately twofold. While the ultrastructure of liver cells remained unaltered after drug treatment, kidney cells exhibited cytoplas-mic vacuolization, membraneous disruption and especially mitochondrial alterations. In accordance with the changes in the ultrastructure of Opossum cells, was the reorganization of cytoskeletal elements with an increased stress fiber network at the basolat-eral surface as well as a partial depolymerization of microtubules and vimentin filaments. A cytoskeletal reorganization was not observed for Chang liver cells after SMCA treatment.

4 The results demonstrate that SMCA causes a dose-dependent cytotoxicity which is accompanied by metabolic, mitochondrial and cytoskeletal alterations in the cells.

Keywords

sodium monochloroacetate cytotoxicity lactate pyruvate mitochondria cytoskeleto

References

1 Woodard G , Lange SW , Nelson KW , Calvery HO . The acute oral toxicity of acetic, chloroacetic, dichloroacetic and trichloroacetic acids . Journal of Industrial Hygiene and Toxicology 1941; 23: 78-82 .

2. Sax N , Lewis R . (1987). Hawley's Condensed Chemical Dictionary. 11th edn. Nostrand Reinhold Company: New York .

3. Feldhaus K , et al. Pediatric fatality associated with accidental oral administration of monochloroacetic acid (MCA) . Veterinary and Human Toxicology 1993; 35: 344-344 .

4. Kulling P , et al. Fatal systemic poisoning after skin exposure to monochloroacetic acid . Clinical Toxicology 1992; 30: 643-652 .

5. Kusch GD , McCarty LP , Lanham JM . Monochloroacetic acid exposure: a case report . Polish Journal of Occupational Medicine 1990; 3: 409-414 .

6. Rogers DR . Accidental fatal monochloroacetic acid poisoning . American Journal of Forensic Medicine and Pathology 1995; 16: 115-116 .

7. Christiansen M , Dalgaard-Mikkelsen S . Toxic effect of monochloroacetate on geese . Acta Pharmacologica et Toxicologica 1961; 18: 179-182 .

8. Quick MP , Manser PA , Stevens H , Bolton JF . Sodium monochloroacetate poisoning of cattle and sheep . Veterinary Record 1983; 8: 155-156 .

9. Quick MP , Porter S , Bennett AD , Fleetwood AJ . Sodium monochloroacetate poisoning of greenfinches . Forensic Science 1992; 54: 1-8 .

10.

10. Christman RF , Norwood DL , Millington DS , Johnson JD . Identity and yields of major halogenated products of aquatic fulvic acid chlorination . Environmental Science and Toxicology 1983; 17: 625-628 .

11.

11. Coleman WE , et al. Gas chromatography/mass spectroscopy analysis of mutagenic extracts of aqueous chlorinated humic acid. A comparison of the by-products to drinking water contaminants . Environmental Science and Toxicology 18: 674-681 .

12.

12. Krasner SW , et al. The occurrence of disinfection by-products in U.S. drinking water . Journal of the American Water Works Association 1989; 81: 41-53 .

13.

13. Uden PC , Miller JW . Chlorinated acids and chloral in drinking water . Journal of the American Water Works Association 1983; 75: 524-527 .

14.

14. Reimann S , Grob K , Frank H . Chloroacetic acids in rainwater . Environmental Science and Toxicology 1996; 30: 2340-2344 .

15.

15. Liebler DC , Meredith MJ , Guengerich FP . Formation of glutathione conjugates by reactive metabolites of vinylidene chloride in microsomes and isolated hepatocytes . Cancer Research 1985; 45: 186-193 .

16.

16. Rannug U , Gothe R , Wachtmeister CA . The mutagenicity of chloroethylene oxide, chloroacetaldehyde, 2-chloroethanol and chloroacetic acid, conceivable metabolites of vinyl chloride . Chemico-Biological Interactions 1976; 12: 251-263 .

17.

17. Yllner S . Metabolism of 1,2-dichloroethane-'4C in the mouse . Acta Pharmacologica et Toxicologica 1971; 30: 257-265 .

18.

18. Yllner S . Metabolism of 1,1,2-trichloroethane-1,2-14C in the mouse . Acta Pharmacological et Toxicologica 1971; 30: 248-256 .

19.

19. Yllner S . Metabolism of chloroacetate in the mouse . Acta Pharmacologica et Toxicologica 1971; 30: 69-80 .

20.

20. Bhat HK , Ahmed AE , Ansari GAS . Toxicokinetics of monochloroacetic acid: a whole-body autoradiography study . Toxicology 1990; 63: 35-43 .

21.

21. Hayes FD , Short RD , Gibson JE . Differential toxicity of monochloroacetate, monofluoroacetate and monoiodoacetate in rats . Toxicology and Applied Pharmacology 1973; 26: 93-102 .

22.

22. Koyama H , et al. Establishment and characterization of a cell line from the American opossum (Didelphis virginiana) . In Vitro 1978; 14: 239-246 .

23.

23. Mossman T . Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays . Journal of Immunological Methods 1983; 65: 55-63 .

24.

24. Borenfreund E , Puerner J . Toxicity determined in vitro by morphological alterations and neutral red absorption . Toxicology Letters 1985; 24: 119-124 .

25.

25. Dartsch PC , Ritter M , Haussinger D , Lang F . Cytoskeletal reorganization in NIH 3T3 fibroblasts expressing the ras oncogene . European Journal of Cell Biology 1994; 63: 316-325 .

26.

26. Gutmann I , Wahlefeld AW . L-(+)-Lactat-Bestimmung mit Lactat-Dehydrogenase und NAD. In Bergmeyer HU (ed) Methoden der enzymatischen Analyse, Bd. II, 3. Auflage. Verlag Chemie: Weinheim , 1974; pp 1510-1514.

27.

27. Czok R , Lamprecht W . Pryvuat, Phosphoenolpyruvat und D-Glycerat-2-phosphat. In Bergmeyer HU (ed) Methoden der enzymatischen Analyse, Bd. II, 3. Auflage. Verlag Chemie: Weinheim , 1974; pp 1491-1496.

28.

28. Dartsch PC . Mounting media for epifluorescence microsopy. A comparative study on the cytoskeleton of cultivated lens epithelial cells . GIT Labormedizin 1990; 13: 450-456 .

29.

29. Dartsch PC , et al. Effects of calcium channel blockers on NIH 3T3 fibroblasts expressing the Ha-ras oncogene . European Journal of Cell Biology 1995; 67: 372-378 .

30.

30. Morrison JL . Toxicity of certain halogen substituted aliphatic acids for white mice . Journal of Pharmacology and Experimental Therapeutics 1946; 86: 336-338 .

31.

31. Bruschi SA , Bull RJ . In vitro cytotoxicity of mono, di-and trichloroacetate and its modulation by hepatic peroxisome proliferation . Fundamental and Applied Toxicology 1993; 21: 366-375 .

32.

32. Giller S , Le Curieux F , Erb F , Marzin D . Comparative genotoxicity of halogenated acetic acids found in drinking water . Mutagenesis 1997; 12: 321-328 .

33.

33. Mitroka JG. (1989). Monochloroacetic acid lethality in the rat in relation to lactic acid accumulation in the cerebrospinal fluid. Dissertation, Rutgers State University of New Jersey, No. 9008948.

34.

34. Millischer RJ , et al. (1987). Monochloroacetic acid: seven worldwide cases of systemic poisoning resulting from accidental skin contact. Occupational Health in Chemical Industry. 22th ICOH-Congress, Sydney , MEDICHEM 1988; 138-144.

35.

35. Lieberg C , Peters RA . The toxicity of fluoroacetate and tricarboxylic acid cycle . Biochimica Biophysica Acta 1949; 3: 215-230 .

36.

36. Bosakowski T , Levin AA . Serum citrate as a peripheral indicator of fluoroacetate and fluorocitrate toxicity . Toxicology and Applied Pharmacology 1986; 85: 428-436 .

37.

37. Bryant BJ , et al, Toxicity of monochloroacetic acid administered by gavage to F344 rats and B6C3F1 mice for up to 13 weeks . Toxicology 1992; 72: 77-87 .

38.

38. Ghadially FN . Ultrastructure and Pathology of the Cell and Matrix. Butterworths: London, Boston, Sydney, Wellington, Durban, Toronto , 1982; pp 196-203.