RESPONSE OF FIBROBLASTS, MAST CELLS AND LIVER COLLAGEN ON FORMATION OF EXPERIMENTAL AMYLOIDOSIS

Abstract

The aim of the study was to quantify the response of fibroblasts, mast cells and liver collagen to formation of experimental amyloidosis. Material and methods. All mice were kept on a standard vivarium diet. Experimental mice underwent formation of amyloidosis by subcutaneous administration of soy cream substitute. Intact mice (IG – 3 specimen) and the internal control group for amyloidosis development (KM – 3 specimen) received water in a free access mode. For mice of the second (VM – 3) and third groups (VFM – 3), water was replaced with dry red grape wine "Cabernet Sauvignon" (Crimea) with an ethyl alcohol content of 10–12°, sugar – 5–10 mg/dm³ against the background of amyloidosis. In addition, fructose 5 g/100 ml of wine was added to the wine of VFM group. To assess the relative area of amyloid-free collagen fibers, dewaxed liver sections were stained using the tricolor technique developed by V.A. Kozlov et al. (2017), to assess the relative area of amyloid deposits and to count fibroblasts (FB) – staining with hematoxylin and Congo red, to identify mast cells (TC) and to assess their functionality – Unna's method. The functional state of mast cells was assessed by the degranulation index (ID) and the sulfatation index (IC) which we proposed earlier. The obtained numerical data were used to calculate a new statistical value – the TC/FB index. The data are presented in the form of mean values and median values indicating the percentiles 10 and 90. Differences of the groups were determined using z‑test. Differences in median values were determined using F-test. Results and their discussion. Liver sections of intact mice corresponded to the histological norm. The relative area of amyloid in the liver sections in CM group was 15.2±2.26%, VM group – 1.9±2.1 and VMF group – 2.5±2.9% (p= 0.0000). The relative area of amyloid–free collagen fibers in IG was 0.50±0.18%, in KM is 0.11±0.03 (p = 0.0151), in VM is 0.51±0.16 and in VMF – 0.69±0.18%. ID in IG – 0.38±0.19, KM – 0.39±0.2, VM – 0.55±0.09, VMF – 1.03±0.19 (p = 0.0065 to IG). IS in IG – 0.57±0.19, KM – 0.38±0.2, VM – 0.54±0.09, VMF – 1.23±0.17 (p = 0.0051 to IG). The median values of TC number in IG were 0.8 (0.5¸1.3), KM – 0.4 (0.3¸0.5), VM – 3.0 (2.0¸3.7), VMF – 2.0 (1.3¸3.0), to IG p < 0.05 in all cases. The median values of the FB number in IG were 35.3 (29.9¸39.9), KM – 40.1 (26.1¸55.4), VM – 28.6 (20.3¸33.7), to IG p < 0.05 in all cases, VMF – 51.3 (46.2¸55.4), p > 0.5. The median values of the TC/FB index in IG were 0.022 (0.013¸0.038), KM – 0.009 (0.007¸0.012), p = 0.0000, VM – 0.11 (0.063¸0.167), p = 0.0012, VMF – 0.037 (0.024¸0.06). Conclusions. When forming experimental amyloidosis in the liver of mice, there is a decrease in the number of TC with a parallel increase in the number of FB per field of vision. Substitution of water in the diet of mice by dry red grape wine inhibits the development of changes characteristic of experimental amyloidosis induced by soy cream substitute. A new statistical value – the TC/FB index reflects both the severity of amyloid liver damage and the effectiveness of preventive measures in the form of substituting water by wine.