Suppression of Gut Bacterial Translocation Ameliorates Vascular Calcification through Inhibiting Toll-Like Receptor 9-Mediated BMP-2 Expression

Figure 4

Antibiotics ameliorate vascular calcification in CRF rats through TLR9 signaling. (a) HASMCs were treated with sham rats’ serum or CRF rats’ serum, with or without TLR4 inhibitor (1 μg/mL, preincubated for 30 min) or TLR9 inhibitor (1 μmol/L, preincubated for 30 min), and then treated with Pi for 7 days. Calcium content was measured (, vs. sham serum + Pi, ^# vs. CRF serum + Pi). (b-d) Male 8-week-old c57 mice, TLR4^-/-, mice and TLR9^-/- mice were fed with high-casein diet (Ctrl) or high-casein diet plus adenine (CRF) for 8 weeks. (b) Aortic rings of c57, TLR4^-/- mice, and TLR9^-/- mice were cultured and treated with sham rats’ serum or CRF rats’ serum and then treated with Pi for 7 days. Calcium content was measured. , vs. c57 + sham serum + Pi, # vs. c57 + CRF serum + Pi. (c) Body weight was measured (~11, , ). (d) Calcium content of abdominal aorta was measured and normalized by dried tissue weight. ~11, vs. c57 Ctrl, # vs. c57 CRF. (e) von Kossa staining of abdominal aorta from c57, TLR4^-/-, and TLR9^-/- mice fed with high-casein diet (Ctrl) or high-casein diet plus adenine (CRF).

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