The reaction of lymphoid splenic tissue of mice on albumin model of systemic amyloidosis (case group, N = 5) was studied and compared to a similar indicator of intact mice (N = 5). Paraffin sections of the spleen, stained with hematoxylin and eosin and Congo red, were microscoped in a regredient LED white light on "Lumam-4" microscope. The absolute area of lymphatic follicles (LFs), their diameters and the area of amyloid lesion were measured on microphotos obtained with the help of video-eyepiece Levenhuk C800 NG 8M in LevenhukLite program. The obtained data were used for calculating the indicators: the relative areas of amyloid lesion (SrelA)), the red (SrelKB) and the white (SrelBP) pulp, the red/white pulp index, LFs' ovalityindex. The number of LFs was counted in the field of vision at magnification of 100. The obtained data were processed using the methods of descriptive and variative statistics and presented in the form of M±m, where M is the mean, m is the standard deviation. Differences of the means were determined using z test. The morphological pattern of the spleen in intact mice corresponded to the histological norm. The wet mass of the spleen in intact animals was 0.75±0.01 g, no signs of amyloidosis were found. In the case group, the wet mass of the spleen increased to 2.2±0.06 g (p=0.000), SrelA was 33.85± 3.39%. The average number of LFs in the field of vision did not change. The diameters by the large and small axes differed by 18% in intact animals and by 6.6% in experimental ones (p=0.000). Respectively, the area of LFs decreased by 11.2% and the ovality index increased by 10.3% (p = 0.0066) in experimental mice. SrelKB and SrelBPdid not change during the formation of amyloidosis. But the Red/White Pulp Index increased by 59.2% (p=0.008). Simulation of amyloidosis in experimental animals was accompanied by a significant increase in the area of the red pulp and by a reduction in the area of white pulp. Thus, the calculated relative morphometric indicators are more informative than the directly measured initial data; the wet mass of the spleen during experimental amyloidosis formation significantly increases; the lymphoid tissue of the spleen readily responds to amyloidogenesis by the change in the ratio of the red and white pulp, as well as by the change in the shape and the area of the lymph follicles.
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.
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