Heparan sulfate and syndecan-1 are essential in maintaining murine and human intestinal epithelial barrier function - PubMed (original) (raw)

Figure 6. Heparin and 2/3-DS-H alleviate protein leakage in vitro and in mice.

(A) Albumin leakage (mean ± SD) through HT29 monolayers relative to untreated cells. Maximum leakage (white bar) was induced by incubating cells with heparinase (HS loss), IFN-γ (10 ng/ml, 24 h), and TNF-α (2 ng/ml, 12 h). Cytokines were coincubated with heparin-like compounds or other GAG derivatives at 2.5 μg/ml or 25.0 μg/ml. Heparin (lane 2) and 2/3-DS-H (lane 9) were most effective in alleviating cytokine-induced protein leakage (arrows). Lane 1, HS; lane 2, high-molecular-weight heparin (unfractionated); lane 3, low-molecular-weight heparin; lane 4, sized heparin, dp 2; lane 5, sized heparin, dp 8; lane 6, sized heparin, dp 14; lane 7, sized heparin, dp 20; lane 8, 2,6-de-_O_-sulfated heparin; lane 9, 2/3-DS-H; lane 10, 6-_O_-desulfated heparin (chemical desulfation); lane 11, 6-_O_-desulfated heparin (enzymatic desulfation with endosulfatase [HSulf2]); lane 12, carboxyl-reduced heparin; lane 13, fully _N_-acetylated heparin; lane 14, fully _O_-sulfated _N_-acetylated heparin; lane 15, chondroitin sulfate; lane 16, fully _O_-sulfated chondroitin sulfate; lane 17, dermatan sulfate; lane 18, fully _O_-sulfated dermatan sulfate; lane 19, fully _O_-sulfated hyaluronic acid; lane 20, archaran sulfate; lane 21, sulfated cyclodextran; lane 22, sucrose octasulfate. (B–G) Intestinal protein leakage in Sdc1+/+ (B, C, E, and F) and _Sdc1_–/– mice (D and G) assessed by in vivo 51Cr labeling. Mice were injected daily with low (100 U/kg, 0.7 mg/kg) (B and E) or high doses (500 U/kg, 3.5 mg/kg) of heparin (C and F), 2/3-DS-H (C, D, F, and G), or PBS as control. Three days following the first injections (t = 0), intestinal protein leakage was induced by injection of either TNF-α (i.v., 0.1 mg/kg) (B–D) or IFN-γ (i.v., 0.2 mg/kg) and TNF-α (i.v., 0.1 mg/kg, 12 h after IFN-γ) (E–G). PBS was used as a control. All data represent assessment in a minimum of n = 4 mice (mean ± SD).