SU1462193A1 - Method of analyzing toxicity of lysosomotropic chemical substances - Google Patents

Abstract

The invention relates to biochemistry. The purpose of the invention is, simplification and. acceleration of the way. The lysosome fraction is incubated in a 0.7 M sucrose solution at 45-50 ° C and the change in optical density is recorded before and after the test substance is added to the lysosome incubation medium. If the value of the ratio of the first indicator to the second is greater than 1, then the test substance has a protective lysosomotropic action. 2 tab.

Description

one. .

The invention relates to medicine, namely to methods for determining the toxicity of chemical substances.

The purpose of the invention is to simplify the method and accelerate the method.

The goal is achieved through the selection of physicochemical factors, the cumulative effect of which causes the necessary degree of labilization by lysosome, i.e. about 50%. The interval values indicated in the formula of factors are chosen in view of this circumstance. Thus, a decrease in the concentration of sucrose solution causes an increase in the degree of labybies of lysosomes (Table 1), and a decrease in temperature — a decrease (Table 2).

Example1. Intact male outbred white rats weighing 120-180 g after 48 h of starvation were used in the work. The liver was taken from decapitated animals, washed on ice with ice-cold saline, blotted with paper filters, thoroughly cleaned of connective tissue, and weighed. The enriched lysosome fraction was obtained by differential centrifugation. A sucrose solution containing 1 mM EDTA was prepared on phosphate-citrate buffer, the pH of which was adjusted to 7 with concentrated NaOH solution. Studies have been conducted on the effect of cadaverine on the stability of intact isolated lysosomes.

The sample was incubated for; - 5 min in 0.7 M sucrose solution pH 7 at 50 s. The decay rate of the li-zosome V, relative to the control value, is 17.0 (D o / min) 10. Cadaverine was added to the sample, until con. concentration 1 mm and incubated in the same way. The lysosome V lysis rate was 4.5 (.). The ratio V, / V is about 4, i.e. more than 1. Therefore, the investigated substance has a protective

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lysosomotropic action and not toxic in this regard for the body.

Example 2. The method is performed as in Example 1. Instead of cadaverine, sodium deoxycholate was added to the sample to a concentration of 0.1%. The degree of labilization of lysosomes was 76%. The ratio of the degree of labilization of these particles during incubation under the conditions specified in Example 1 to the degree of labilization caused by sodium deoxycholate 55% / 76% is less than 1,

Consequently, this | 1 st substance causes the destruction of lysosomes and has a toxic effect on the body when it enters the cells of the tissue.

Example 3. The method is carried out as in Example 1. Instead of cadaverine, sodium dodecyl sulfate was added to the sample to a final concentration of 0.1%. - The degree of labilization was close to 100% (95%). The ratio of 55% / 95% is less than 1. Therefore, this substance is also, like the previous one, toxic to the animal's body.

Table

The effect of the molarity of the sucrose solution on the stability of lysosomes.

Note: Samples are incubated at 50 ° C in a sucrose solution of the indicated molarity pH 7 for 5 minutes.

Table 2 The effect of temperature of the incubation medium on the stability of isolated lysosomes.

Claims (1)

Formula isotopic The method of determining the toxicity of lysosomotropic chemicals by examining the physicochemical properties of lysosomes, characterized in that, in order to simplify and accelerate the method, the stability of schizosomes is investigated by incubating them in 0.7 M sucrose at an incubation temperature of 45-50 ° C, with This records the change in optical density of the resulting suspension per minute before and after adding the lotuses to the incubation medium with the test substance, and when the ratio of the first indicator to the second is less than unity, toxicity is determined. the awn Note: Samples are incubated at the indicated temperature in sucrose plant with 0.7 M pH 7 for 5 minutes.