Assessment of oxidative stress index in patients with non-alcoholic fatty liver disease using spectroscopy and Immunoassay

Abstract

In the process of oxidative stress, fats are oxidized by reactive oxygen species, replacing hydrogen atoms with molecular oxygen, and eventually the formation of low molecular weight oxidized products in the blood, such as Malondialdehydehyde (MDA), which is a non-invasive and vital indicator of the oxidative stress process.

Objective: Comparison of accuracy and sensitivity in the detection of very low concentrations of malon dialdehyde MDA using two analytical methods, the enzyme linked immunoassay) ELIZA( method,  forming a yellow malondialdehyde derivative measured by a microplate reader, and the method of reaction with thiobarbityric acid (TBA), forming a pink malondialdehyde derivative measured spectrally, and then finding the relationship with non-alcoholic fatty liver disease.

Methods: - The study were applied to 90 patients suffering from fatty liver( a 44 males and 46 females). Based on the degree of steatosis, they were divided into three groups: (n=29) healthy, (n=29) mild steatosis and (n=32) moderate to severe D2 steatosis. Serum was collected after it’s  processing for detection malondialdehyde (MDA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), albumin, total bilirubin and vitamin "D". Body mass index (BMI) was calculated by the ratio of weight to height (in kilograms/square of length in meters).

Study results: The results of the comparison between the two analytical methods showed significant statistical differences (p < 0.0001), and according to the quantitative analyses of the blank solution, the detection limit was respectively (1.5 x 10-3 nmol/ml) (2.9 x 10-6 nmol/ml) and quantitative estimation (4.5 x 10-3 nmol/ml) (8.8 x 10-5 nmol/ml). Quantitative analyses showed different trends when patients compared to healthy controls. There was a high trend in the concentration of MDA (p < 0.05), ALT and AST, especially in males, but a low trend in the concentration of total bilirubin with increasing body mass index, especially in females , but the concentration of both vitamin D, ALP and albumin was not statistically significant (p>0.05).

The results of this study confirm the active role of AST, ALT, total bilirubin and BMI in the development of non-alcoholic fatty liver disease,  while the enzyme ALP and albumin and vitamin D was not associated  with non-alcoholic fatty liver disease. The results were in the spectroscopy method with a linear rate  of 99.93%, although it is the least accurate and sensitive, but it is the most specific to detect MDA compared to the immunoassay method (ELIZA), thus highlights the Influential MDA relationship in non-alcoholic fatty liver disease.