Table 1.
Metabolic characteristics of STZ-induced diabetic mice after treatment with PAI-1 inhibitors.
Fig 1.
TM compounds improve kidney function and morphology in STZ-induced diabetic mice.
Diabetes was induced in mice by intraperitoneally injecting STZ (150 mg/kg), and then either TM5275 (50 mg/kg/day) or TM5441 (10 mg/kg/day) was administered orally for 16 weeks to the diabetic and age-matched control mice. After 16 weeks, blood was collected for analysis of (A) plasma creatinine, and urine was collected for (B) protein analysis by using the Bradford assay and SDS-PAGE. (C) Kidneys were fixed in paraffin and cut into 4-μm sections that were subsequently stained with PAS (periodic acid–Schiff) reagent. Scale bar: 10 μm; original magnification: 630×. After PAS staining, (D) glomerular volume and (E) FMA were analyzed using Image-Pro Plus 4.5.1. CM, control mice; DM, STZ-induced diabetic mice. Data are presented as the mean ± SE of 5–8 mice/group; *p < 0.05 vs. CM, †p < 0.05 vs. DM.
Fig 2.
TM compounds inhibit kidney fibrosis in STZ-induced diabetic mice.
After diabetic mice were treated with TM compounds for 16 weeks, the mRNA and protein levels of fibrosis markers in kidney tissue were measured. (A) Collagen Iα1, (B) fibronectin, and (C) α-SMA mRNA expression levels were measured using real-time PCR. Paraffin-embedded kidney sections were stained with (D) Masson trichrome stain (original magnification: 630×; scale bar: 10 μm), (E) picrosirius red (original magnification: 200×; scale bar: 100 μm), or anti-fibronectin antibodies; original magnification: 630×; scale bar: 10 μm). CM, control mice; DM, STZ-induced diabetic mice. Data are presented as the mean ± SE of 5–8 mice/group; *p < 0.05 vs CM, †p < 0.05 vs DM.
Fig 3.
TM compounds inhibit kidney inflammation in STZ-induced diabetic mice.
After diabetic mice were treated for 16 weeks with TM compounds, mRNA and protein expression levels of inflammatory cytokines were measured in the kidney tissue. Real-time PCR analysis of the mRNA expression of (A) MCP-1, (B) F4/80, and (C) PAI-1. (D, E) Paraffin-embedded kidney sections were stained with anti-F4/80 antibodies (1:200; original magnification: 630×; scale bar: 10 μm). CM, control mice; DM, STZ-induced diabetic mice. Data are presented as the mean ± SE of 5–8 mice/group; *p < 0.05 vs CM, †p < 0.05 vs DM.
Fig 4.
TM compounds inhibit PAI-1-induced fibrotic and inflammatory responses in vitro.
We treated mProx cells with TM compounds for 4 h and then stimulated them with 50 nM recombinant PAI-1 for 24 h. Real-time RT-PCR was used to measure the mRNA expression of (A) TGF-β1, (B) collagen Iα1, (C) collagen Iα2, and (D) MCP-1. (E) Plasmin activity was also measured. Data are presented as the mean ± SE of 4 experiments; *p < 0.05 vs control, †p < 0.05 vs PAI-1.