Journal: Scientific Reports

Article Title: Splenectomy reduces shear stress and inflammation in liver endothelial cells during regeneration after partial hepatectomy in mice

doi: 10.1038/s41598-025-32446-4

Figure Lengend Snippet: Characteristics of liver regeneration after 70% resection under conditions of previous splenectomy. ( A ) Histological structure of the regenerating liver, hematoxylin and eosin stain, scale bar—100 µm, arrows indicate mitotic figures in hepatocytes. ( B ) Dynamics of liver mass recovery after 70% resection. ( C ) Serum concentrations of ALT, AST and albumin in operated mice. ( D ) Immunohistochemical study of the proliferation marker Ki67 in the regenerating liver. Second antibodies are conjugated to FITC (green), nuclei are counterstained with DAPI (blue), scale bars—50 μm. ( E ) Dynamics of the Ki67 index. Data are presented as mean ± SD, SO – sham splenectomy + sham hepatectomy (n = 6), SE—splenectomy, (n = 6), SH – splenectomy + hepatectomy (n = 18), PH—sham splenectomy + hepatectomy, *—statistically significant differences, p < 0,05. Paired comparisons used the Student’s t-test for normal distributions and the Mann–Whitney U-test for distributions other than normal.)

Article Snippet: A 10 5 aliquote of LSECs isolated using magnetic sorting for the CD146 marker were incubated in 100 μl Rinsing Solution (Miltenyi Biotec, USA) with 5 μl primary antibodies to CD31 (PECAM-1) (PE-labeled, 130–111-540, clone REA784 | 390, Miltenyi Biotec, USA), integrin alpha-5 (CD49e) (PE-labeled, 130–122-072, clone REA1183 | 5H10-27, Miltenyi Biotec, USA), VCAM-1 (CD106) (PE-labeled, 130–116-323, clone REA971 | 429, Miltenyi Biotec, USA), VE-cadherin (CD144) (PE-labeled, 130–128-207, clone REA225 | BV13, Miltenyi Biotec, USA), Ki67 (FITC-labeled, 130–117-691, clone REA183 | B56 Miltenyi Biotec, USA), CD146 (130–102-230, clone ME-9F1, Miltenyi Biotec, USA), F4/80 (130–102-422, REA126 | BM8, Miltenyi Biotec, USA ) at room temperature for 1 h. The dynamics of CD3 + lymphocytes and NK1.1 cells in regenerating livers were analyzed similarly using 10 5 cells of the liver stromal fraction and primary antibodies to CD45 (PE-labeled, 130–102-596, clone 30F11, Miltenyi Biotec, USA), CD3 (APC-labeled, MCA500APC, clone KT3, Bio-Rad USA), NK1.1 (VioBlue-labeled, Biolegend, USA); the incubations proceeded for 1 h. Following the incubations, the cells were washed in PBS, resuspended in 0.5 ml PBS and analyzed in a MACSQuant 10 flow cytometer (Milteniy Biotech, Germany); the data were analyzed in FlowJo (LLC).

Techniques: H&E Stain, Immunohistochemical staining, Marker, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Splenectomy reduces shear stress and inflammation in liver endothelial cells during regeneration after partial hepatectomy in mice

doi: 10.1038/s41598-025-32446-4

Figure Lengend Snippet: Activity of cell death and proliferation of endotheliocytes of sinusoidal capillaries of regenerating liver. ( A ) Dynamics of cell death in the regenerating liver. ( B ) Dynamics of the population of Ki67 + endotheliocytes in the regenerating liver. Data are presented as mean ± SD, SO – sham splenectomy + sham hepatectomy (n = 6), SE—splenectomy, (n = 6), SH – splenectomy + hepatectomy (n = 18), PH—sham splenectomy + hepatectomy, *—statistically significant differences, p < 0,05. Paired comparisons used the Student’s t-test for normal distributions and the Mann–Whitney U-test for distributions other than normal.

Article Snippet: A 10 5 aliquote of LSECs isolated using magnetic sorting for the CD146 marker were incubated in 100 μl Rinsing Solution (Miltenyi Biotec, USA) with 5 μl primary antibodies to CD31 (PECAM-1) (PE-labeled, 130–111-540, clone REA784 | 390, Miltenyi Biotec, USA), integrin alpha-5 (CD49e) (PE-labeled, 130–122-072, clone REA1183 | 5H10-27, Miltenyi Biotec, USA), VCAM-1 (CD106) (PE-labeled, 130–116-323, clone REA971 | 429, Miltenyi Biotec, USA), VE-cadherin (CD144) (PE-labeled, 130–128-207, clone REA225 | BV13, Miltenyi Biotec, USA), Ki67 (FITC-labeled, 130–117-691, clone REA183 | B56 Miltenyi Biotec, USA), CD146 (130–102-230, clone ME-9F1, Miltenyi Biotec, USA), F4/80 (130–102-422, REA126 | BM8, Miltenyi Biotec, USA ) at room temperature for 1 h. The dynamics of CD3 + lymphocytes and NK1.1 cells in regenerating livers were analyzed similarly using 10 5 cells of the liver stromal fraction and primary antibodies to CD45 (PE-labeled, 130–102-596, clone 30F11, Miltenyi Biotec, USA), CD3 (APC-labeled, MCA500APC, clone KT3, Bio-Rad USA), NK1.1 (VioBlue-labeled, Biolegend, USA); the incubations proceeded for 1 h. Following the incubations, the cells were washed in PBS, resuspended in 0.5 ml PBS and analyzed in a MACSQuant 10 flow cytometer (Milteniy Biotech, Germany); the data were analyzed in FlowJo (LLC).

Techniques: Activity Assay, MANN-WHITNEY

Journal: Scientific Reports

Article Title: The local anesthetic, lidocaine, suppresses the growth of colon cancer SW480 cells by decreasing the voltage-gated sodium channel subunit Na v 1.5

doi: 10.1038/s41598-025-29505-1

Figure Lengend Snippet: Lidocaine induced growth suppression along with a decrease of Na v 1.5 levels. ( A ) Lidocaine treatment induced growth suppression of SW480 cells in a dose-dependent manner. SW480 cells were treated with the indicated concentration of lidocaine for 24 h, and cell proliferation assay was performed. The lysate of SW480 cells treated with the indicated concentration of lidocaine was subjected to western blot analysis. The arrow indicates Na v 1.5. The expression levels of Na v 1.5 were normalized to α-tubulin levels in each sample. The results of each sample assayed in triplicate are presented as the mean ± SD. * p < 0.05, ** p < 0.01. ( B ) Lidocaine treatment decreased Na v 1.5 expression in the cell membrane of SW480 cells, along with a decrease of the proliferation marker, Ki-67. SW480 cells untreated or treated with 3 mM lidocaine for 24 h were applied for immunofluorescence assay. The nuclei were stained with DAPI. The arrows indicate Na v 1.5 localized in the cell membrane of SW480 cells. The scale bar represents 10 μm. ( C ) Lidocaine induced growth suppression of SW480 cells with a decrease of Ki-67 expression. For quantification of Ki-67 expression in SW480 cells, cells untreated or treated with 3 mM lidocaine for 24 h were applied for immunofluorescence assay and were stained with anti-Ki67 antibody. The nuclei were stained with DAPI. The scale bar represents 10 μm. The number of Ki-67 positive and negative SW480 cells with or without lidocaine treatment are shown. Each sample was assayed in triplicate and the results are presented as the mean ± SD. * p < 0.05.

Article Snippet: The antibodies used in this study were rabbit anti-Na v 1.5 antibody (Proteintech, IL, USA, 23016-1-AP), mouse anti-Ki67 antibody (SantaCruz Bio, TX, USA, sc-23900), rabbit anti-ATP1A1 antibody (Proteintech, IL, USA, 55187-1-AP), rabbit anti-GDF-15 antibody (Proteintech, IL, USA, 27455-1-AP), rabbit anti-cyclin D1 antibody (Proteintech, IL, USA, 26929-1-AP), mouse anti-p53 antibody (Cell Signaling Technology, MA, USA, #2524), anti-rabbit cleaved caspase-3 antibody (Cell Signaling Technology, MA, USA, #9664), mouse anti-GAPDH antibody (Fujifilm-Wako, Osaka, Japan, 014-25524), mouse α-tubulin antibody (DM1A) (SantaCruz Bio, TX, USA, sc-32293), goat Cy3-conjugated anti-mouse IgG antibody, and donkey Cy2-conjugated anti-rabbit IgG antibody (Jackson ImmunoResearch Laboratories, PA, USA).

Techniques: Concentration Assay, Proliferation Assay, Western Blot, Expressing, Membrane, Marker, Immunofluorescence, Staining