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

Figure 3

Bacterial components LPS and DNA promote Pi-induced calcification and osteoblastic differentiation in HASMCs. HASMCs were treated with inorganic phosphate (Pi), E. coli ssDNA (10 μg/mL), or LPS (100 ng/mL) as indicated for 7 days, and alizarin red staining was operated. (b-d) HASMCs were treated with TLR9 ligand control (1 μg/mL) or E. coli ssDNA (ssDNA, 10 μg/mL), with or without Pi for 7 days. (b) Calcium content was measured. Real-time PCR analyzed the mRNA levels of (c) Cbfα1 and (d) SM22α. Data are relative to β-actin level. (e-g) HASMCs were treated with LPS at the indicated concentrations (ng/mL), with or without Pi for 7 days. (e) Calcium content was measured. Real-time PCR analyzed the mRNA levels of (f) Cbfα1 and (g) SM22α. Data are relative to β-actin level. , vs. Ctrl, ^# vs. Pi.

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