Enhanced expression of ETRs in spleen tissues of patients with schistosome-induced hepatic fibrosis.
(A) The human spleen tissues were stained with H&E and Masson’s trichrome. Scale bar, 200 μm. (B-G) qPCR analysis of the expression levels of ET-1, ETAR, ETBR, Col1α1, Col3α1 and IL-10 in spleen samples. Cont (n = 9), CS (n = 12), CHB (n = 13). (H) The IL-10 concentration in spleen tissue homogenates was determined by ELISA. Cont (n = 9), CS (n = 12), CHB (n = 13). (I) Representative immunohistochemical staining of ETAR, ETBR, collagen I and collagen III. Black arrows indicate the ETRs, collagen I and collagen III positive cells. Scale bar, 100 μm. (J) Representative immunofluorescence staining of ETAR, ETBR and CD20+ B cells in human spleen tissues. Insets show a higher magnification of the outlined area. Yellow arrows denote positive cells. Scale bar, 50 μm. (K) The fluorescence intensity was measured using IPP software (n = 4). (L-N) ETAR and ETBR proteins were determined by western blotting, quantified using Image J analysis, and normalized to GAPDH (n = 7). Data are represented as the median and interquartile range from two independent experiments. Multiple comparisons were performed by Kruskal–Wallis with Dunn’s posttest (B-H, M-N) or the Mann-Whitney U-test (K). *P < 0.05. Abbreviation: Cont, control; CS, chronic schistosomiasis; CHB, chronic hepatitis B; RP: red pulp; WP: white pulp.