Bairati C, Goi G, Lombardo A, Tettamanti G
Department of Medical Chemistry and Biochemistry, Medical School University of Milan, Italy.
Clin Chim Acta. 1994 Jan 31;224(2):147-57. doi: 10.1016/0009-8981(94)90181-3.
An in vitro test was set up to assess the release of lysosomal enzymes from cells and the effect on this process of the commonly used preservatives, parabens. Human peripheral lymphocytes, cultivated in vitro for 24 h in the presence or absence of phytohaemagglutinin (PHA; 5 mg/l), were used. After 1 day of incubation, PHA treatment caused an increased release (from 220 to 500%) of the lysosomal enzymes N-acetyl-beta-D-glucosaminidase, beta-D-glucuronidase, alpha-L-fucosidase and alpha-D-galactosidase. This enhancement was analytically reliable, and detectable with 1-5 micrograms of cell protein. Leakage of lactate dehydrogenase (LDH) underwent only a 20% increase on PHA treatment, indicating that the increased release of lysosomal enzymes was presumably due to secretion, not to cell damage. In PHA-stimulated lymphocytes, methyl-, ethyl-, propyl- and butyl-parabens caused a concentration-dependent diminution of the secretion of lysosomal enzymes. Butyl-paraben appeared to be the most potent inhibitor, causing a 45-50% inhibition at 0.06 mmol/l. With the other parabens, the inhibitory effect became statistically significant at about 0.25 mmol/l for alpha-L-fucosidase and alpha-D-galactosidase, and at 0.5 mmol/l for N-acetyl-beta-D-glucosaminidase and beta-D-glucuronidase. At 1 mmol/l inhibition was greater than 50% for all the enzymes and was more marked with the propyl derivative. Parabens did not influence the release of LDH, suggesting that they affected particularly the secretion of lysosomal enzymes. This supports the hypothesis that parabens are capable of affecting cellular function at concentrations which are likely to be reached in blood or tissues under conditions of common use.
开展了一项体外试验，以评估细胞溶酶体酶的释放情况以及常用防腐剂对羟基苯甲酸酯对此过程的影响。使用在存在或不存在植物血凝素（PHA；5 mg/l）的情况下体外培养24小时的人外周血淋巴细胞。孵育1天后，PHA处理导致溶酶体酶N-乙酰-β-D-氨基葡萄糖苷酶、β-D-葡萄糖醛酸酶、α-L-岩藻糖苷酶和α-D-半乳糖苷酶的释放增加（从220%增至500%）。这种增强在分析上是可靠的，并且在1-5微克细胞蛋白时即可检测到。PHA处理后，乳酸脱氢酶（LDH）的泄漏仅增加了20%，这表明溶酶体酶释放增加可能是由于分泌，而非细胞损伤。在PHA刺激的淋巴细胞中，甲基、乙基、丙基和丁基对羟基苯甲酸酯导致溶酶体酶分泌呈浓度依赖性减少。丁基对羟基苯甲酸酯似乎是最有效的抑制剂，在0.06 mmol/l时可产生45-50%的抑制作用。对于其他对羟基苯甲酸酯，α-L-岩藻糖苷酶和α-D-半乳糖苷酶在约0.25 mmol/l时抑制作用具有统计学意义，N-乙酰-β-D-氨基葡萄糖苷酶和β-D-葡萄糖醛酸酶在0.5 mmol/l时抑制作用具有统计学意义。在1 mmol/l时，所有酶的抑制率均大于50%，丙基衍生物的抑制作用更明显。对羟基苯甲酸酯不影响LDH的释放，表明它们特别影响溶酶体酶的分泌。这支持了这样一种假设，即在常用条件下，对羟基苯甲酸酯能够在血液或组织中可能达到的浓度下影响细胞功能。
