Table 1.
Experimental designs and protocolsa.
Table 2.
Disease activity index (DAI) for clinical evaluation of DSS-induced UC in micea.
Table 3.
Histopathological scoring systema.
Fig 1.
Effects of MSE on DAI scores in DSS-induced UC.
(A) Acute UC (n = 8/group): data are shown at daily intervals throughout UC induction and treatment periods. (B) Chronic UC (n = 6/group): data are shown at 3-d intervals during UC induction and 2-d intervals during the treatment period. Experimental designs and disease induction procedures are defined in Table 1 and DAI scoring criteria in Table 2. Data are expressed as mean ± SEM. *p<0.05, **p<0.01, and ***p<0.001 compared to disease control (DSS/vehicle). 5-aminosalicylic acid (5-ASA); disease activity index (DAI); dextran sulfate sodium (DSS); moringa seed extract (MSE); ulcerative colitis (UC).
Fig 2.
Effects of MSE on spleen and colon in DSS-induced UC.
(A) Spleen weight (mg), (B) colon length (cm), and (C) colon weight/length ratio (mg/cm) were measured at necropsy of acute UC (n = 8/group) and chronic UC (n = 6/group) experiments. Experimental designs and disease induction procedures are defined in Table 1. Data are expressed as mean ± SEM. Different letters (a, b, c) indicate significant differences between groups at p<0.05. 5-aminosalicylic acid (5-ASA); dextran sulfate sodium (DSS); moringa seed extract (MSE); ulcerative colitis (UC).
Fig 3.
Effects of MSE on colon histopathology in DSS-induced UC.
Representative histological observations of H&E-stained colonic sections of (A) acute UC mice (n = 8/group), and (B) chronic UC mice (n = 6/group). Experimental designs and disease induction procedures are defined in Table 1. (C) Associated colonic histopathological scores calculated as sum of scores of each parameter consisting of histopathological scoring system as described in Table 3. Histological images were acquired by X20 and X400 magnification, respectively (Scale bar = 25 μm). Data are expressed as mean ± SEM. Different letters (a, b, c) indicate significant differences between groups at p<0.05. 5-aminosalicylic acid (5-ASA); dextran sulfate sodium (DSS); hematoxylin and eosin (H&E); moringa seed extract (MSE); ulcerative colitis (UC).
Fig 4.
Effects of MSE on colonic pro-inflammatory biomarkers and fecal Lcn-2 levels in DSS-induced UC.
Analyses of (A) KC, (B) TNF-α, (C) NO, and (D) MPO were performed in colon culture supernatants of acute UC (n = 8/group) and chronic UC (n = 6/group) mice. All concentrations were normalized to 1 g of the colon tissue weight. Fecal levels of (E) Lcn-2 were normalized to 1 g of the fecal weight. Experimental designs and disease induction procedures are defined in Table 1. Data are expressed as mean ± SEM. Different letters (a, b, c) indicate significant differences between groups at p<0.05. 5-aminosalicylic acid (5-ASA); dextran sulfate sodium (DSS); keratinocyte-derived cytokine (KC); lipocalin-2 (Lcn-2); myeloperoxidase (MPO); moringa seed extract (MSE); nitric oxide (NO); tumor necrosis factor-α (TNF-α); ulcerative colitis (UC).
Fig 5.
Effects of MSE on colonic expression of pro-inflammatory markers, tight-junction proteins, and Nrf2-mediated enzymes in DSS-induced UC.
Gene expression of (A) IL-1β, (B) IL-6, (C) TNF-α, (D) iNOS, (E) claudin-1, (F) ZO-1, (G) GSTP1, (H) NQO1, and (I) HO1 was analyzed by qPCR in colon tissues of acute UC (n = 8/group) and chronic UC (n = 6/group) mice. All data are expressed as a relative fold change compared to healthy control (value = 1.0). Experimental designs and disease induction procedures are defined in Table 1. Data are expressed as mean ± SEM. Different letters (a, b) indicate significant differences between groups at p<0.05. 5-aminosalicylic acid (5-ASA); dextran sulfate sodium (DSS); glyceraldehyde-3-phosphate dehydrogenase (GAPDH); glutathione-S-transferase pi 1 (GSTP1); hydroxymethylbilane synthase (HMBS); heme oxygenase 1 (HO1); interleukin-1β (IL-1β); interleukin-6 (IL-6); inducible nitric oxide synthase (iNOS); moringa seed extract (MSE); NAD(P)H quinone oxidoreductase 1 (NQO1); tumor necrosis factor-α (TNF-α); ulcerative colitis (UC), zonula occludens-1 (ZO-1).