Merjenje vpliva učinkovin na obliko eritrocitov s primerjavo reprezentativnih populacij : diplomsko delo

Vehovar, Urška (author), Kralj-Iglič, Veronika (mentor)

Uvod: Oblika eritrocitov je zaradi odsotnosti notranje strukture določena z lastnostmi eritrocitove membrane, zato se opazovanje oblike eritrocitov pogosto uporablja za raziskave o vplivu kemikalij na celične membrane. Najpogostejša odziva oblike eritrocita na zunanje dejavnike sta ehinocitoza ali stomatocitoza. Namen: Želeli smo izdelati metodo, s katero bi na osnovi spremembe oblike in števila eritrocitov primerjali vpliv učinkovin na celične membrane. Kot primer smo uporabili tri učinkovine, pridobljene s kromatografijo na gelu iz krvi bolnika z Alzheimerjevo boleznijo (označen s številko 1) in dveh zdravih darovalcev (34 in 39). Metode dela: Suspenziji eritrocitov smo dodajali učinkovine in vzorce opazovali pod optičnim mikroskopom takoj po dodatku učinkovine, po eni uri in po 24 urah. Med opazovanjem smo naredili vrsto slik vsakega vzorca, ki smo jo uporabili za analizo oblik in števila celic. Ločili smo stomatocitno, diskocitno, ehinocitno in sferocitno obliko eritrocitov. Vzorce smo primerjali z metodami deskriptivne statistike. Rezultati: V sveži krvi kontrolnih vzorcev so v začetku večinoma diskociti, kasneje pa pride do spontane ehinocitoze in po enem dnevu tudi do hemolize. V vzorcih z dodanima učinkovinama 34 in 39 je bil ta proces hitrejši kot v kontrolnih vzorcih, nastajali so tudi sferociti. Proces je v vzorcu z dodano učinkovino 39 potekal najhitreje. V vzorcih z učinkovino 1 so se diskociti ohranili v največji meri. Po številu celic na sliko se vzorec 34 ni razlikoval od kontrole, vzorec 39 je vseboval manj celic od nje, vzorec 1 pa več. Pri vzorcih z učinkovino 1 je prišlo do sprijemanja celic v skupke. Razprava in sklep: Analiza oblik je pokazala, da lahko ločimo vplive različnih učinkovin na eritrocite od kontrolnih vzorcev in tudi med seboj. Učinkovina pridobljena iz krvi bolnika z Alzheimerjevo boleznijo v fazi remisije je najbolj zavrla spontani potek ehinocitoze v vzorcih. Ugotovili smo tudi nekatere slabosti metode, predvsem pri pripravi opazovalnih komor.

• Language: Slovenian
• Keywords: mikroskopija, ehinociti, statistična analiza, t-test
• Work type: Bachelor thesis/paper
• Organization: ZF - Faculty of Health Sciences
• Year: 2018
• PID: 20.500.12556/RUL-101228
• COBISS.SI-ID: 5428587
• Publication date in RUL: 16.05.2018

Secondary language

• Language: English
• Title: Determination of effect of added substances on erythrocyte shape within representative populations : diploma thesis

Abstract

Introduction: Due to the absence of internal structure, the shape of erythrocytes is determined by erythrocyte membrane properties, therefore, the observation of the erythrocyte form is often used to investigate the effect of chemicals on cell membranes. The most common erythrocyte membrane responses to external factors are echinocytosis and stomatocytosis. Purpose: We wanted to create a method by which, on the basis of a change in the shape and number of erythrocytes, the effect of the substances on the cell membranes would be compared. As an example, three substances derived from the blood of the patient with Alzheimer's disease (labeled by number 1) and two healthy donors (34 and 39) were used. Methods: The substances were added to the erythrocyte suspension and the samples were observed under the optical microscope immediately after the addition of the substance, after one hour and 24 hours. During the observation we made a series of images of each sample that was used to analyze the shape and number of cells. We separated the stomatocytic, discocytic, echinocytic and spherical form of erythrocytes. Samples were compared with descriptive statistics. The results: In the fresh blood of control samples, they are mostly discocytes, later it comes to spontaneous echinocytosis and after one day, even hemolysis. In the samples with added substances 34 and 39, this process was faster than in control samples, and spherocytes were also formed. The process was the fastest in the sample with the added substance 39. In samples with substance 1, the discocytes were longest preserved. According to the number of cells per picture, the sample 34 did not differ from the control, the sample 39 contained fewer cells than it, and the sample 1 more. In the samples with substance 1, the cells were joining in aggregates. Discussion and conclusion: The analysis of the forms has shown that the effects of various substances on erythrocytes can be distinguished from the control samples as well as among themselves. The substance obtained from the blood of a patient with Alzheimer's disease in the remission phase had the greatest effect on the spontaneous course of echinocytosis in the samples. We also found some weaknesses in the method, especially in the preparation of observation chambers.

• Keywords: microscopy, echinocytes, statistical analysis, t-test