FN1 mRNA expression of fibronectin 1 and distribution of fibronectin-associated leukocytes in humans with chronic diffuse liver diseases

Abstract

Chronic diffuse liver diseases are characterized by continuous progression of fibrosis, ultimately leading to cirrhosis with the following loss of the normal functioning of this organ due to excessive accumulation of the components of extracellular matrix. To find new, more available diagnostic markers of detecting disorders in the liver, we used methods of antifungal cytofluorometry and quantitative real-time polymerase chain reaction. Intensity of exposure of fibronectin and plasmatic membrane of lymphocytes in the group of patients with chronic diffuse diseases compared with the control group of practically healthy donors decreased both inside and on the surface of the cells respectively by 45.3% and 16.2%. Similar tendency towards decrease was observed during the assays of the level of the exposure of fibronectin on the surface and inside the blood granulocytes: by 25.0% and 36.5%, respectively. In the blood of the patients suffering from chronic diffuse diseases, compared with the control group, there was determined reliable increase in percentage of lymphocytes and granulocytes which contain topical fibronectin, by 32.3% and 2.78 times, correspondingly. The level of monocytes (as a percentage) with cell-associated fibronectin and fibronectin localized inside, by contrast, reliably decreased in 2.07 and 4.50 times, respectively. Analysis of the expression of FN1 in lymphocytes of blood of the studied groups using quantitative real-time polymerase chain reaction revealed decrease in the level of FN1 mRNA expression by 34.0% in the group of ill patients compared with the control group. We determined excellent diagnostic informativeness of the parameters of the level of exposure of fibronectin inside and on the surface of granulocytes and prognostic accuracy of the classifier from these parameters at the level of 100% using the method of support vector machine, SVM. High levels of diagnostic informativeness were recorded for the tests of all types of analyzed leukocytes with cell-associated fibronectin, and the classifiers based on the pair combinations of the tests with cell-associated fibronectin and fibronectin localized within the cells provide high diagnostic accuracy of the prognosis. Because the mentioned indicators are highly-sensitive tests, they can be proposed for early diagnostics and evaluation of the effectiveness of the conducted therapy of chronic diffuse liver diseases, which would allow reducing the use of paracentetic trepanobiopsy, a painful and risky procedure, which still remains the main type of diagnostic.

Keywords

• fibronectin; fn1 expression; lymphocytes; monocytes; granulocytes.