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. 2020 Jun 5;10(16):7384–7400. doi: 10.7150/thno.45455

Figure 2.

Figure 2

Indoxyl sulfate (IS), a metabolite of intestinal flora, induces intestinal barrier injury. (A) Caco2 cells were seeded on Millicell Hanging Cell Culture Inserts in 24-well plates and cultured for 14 days, and then treated with various concentrations of IS for 24 hours or 500 μM IS for different durations for TER determination. n=3. (B) qPCR analysis of the expression of tight junction-related genes in Caco2 cells treated with 500 μM IS for 24 hours. n=3. (C-H) Mice were intraperitoneally injected with IS (100 mg/kg) daily for 8 weeks. n = 8 per group. HE staining (C) and macroscopic injury score (D, detailed information was shown in Table S5) of intestinal tissues from control and IS-injected mice. (E) Serum IS level was determined using HPLC. Relative intestinal permeability (F), TEM observation of tight junction (G), and qPCR analysis of tight junction-related genes (H) of intestinal tissues from mice in (C). (I) Serum level of TNF-α, IL-1β and IL-6 from mice in (C) were measured using ELISA kits. Yellow arrow indicates intestinal mucosal damage in (C) and impaired tight junction in (G). Data are shown as mean ± SEM and were analyzed by one-way ANOVA (A) or two-tailed unpaired Student's t test (B, D-F, H, I). * P<0.05, ** P<0.01, *** P<0.001.