Figure 6.

MT alleviates vascular calcification and ageing in the 5/6 NTP‐induced mouse model. A, Schematic flow diagram representing the in vivo treatment of MT with or without GW4869 in the 5/6 NTP mouse model. Sham operation mice with high‐phosphate diet were used as control (n = 5 per group). B, Calcium content of the thoracic aorta of mice was measured by o‐cresolphthalein complexone method. C, The ALP activity of thoracic aortic tissues was measured by the specific kit and normalised to total proteins contents. D, Representative microscopic pictures of Von Kossa‐stained and Alizarin Red S‐stained sections from the thoracic aorta (left panel) and quantitation of positive staining area (right panel) are shown. Scare bar 200 µm (Black). E, The expression of RUNX2 in thoracic aorta was examined by immunohistochemistry (IHC) (left panel), and quantitation of positive staining area (right panel) is shown. Scale bar 50 µm (Blue) and 500 µm (Red). F, Representative vascular SA‐β‐gal staining pictures are presented (left panel), and quantitation of positive staining area (right panel) is shown. Scale bar 50 µm (Blue). Green staining area indicates ageing tissues. G, The expression of p21 in thoracic aorta was determined by immunohistochemistry (left panel), and quantitation of positive staining area (right panel) is shown. Scare bar 50 µm (Blue) and 500 µm (Red). Results are represented by mean ± SEM with five replicates for each group. Significance was analysed by two‐way ANOVA with Tukey's HSD post hoc analysis. *P < .05