3-n-Butylphthalide Attenuates Arterial Calcification Induced By Vitamin D3 In Mice

Objective: To observe the effect of 3-n-butylphthalide on arterial calcification in mice and its possible mechanism.Methods: C57BL/6 male mice were randomly divided into control group(N group),vitamin D3 group(VD group),vitamin D3+50mg NBP group(VDN1 group),vitamin D3+100mg NBP group(VDN2group).Except for the control group,vitamin D3(500,000 IU/kg/d)was injected subcutaneously for 7 days to establish a mouse arterial calcification model.From the first day of the experiment,VDN1 group and VDN2 group were given small dose(50mg/kg/d)and large dose(100mg/kg/d)NBP for 14 consecutive days.All mice were killed after feeding for 2 months after the intervention,and the thoracic aorta of mice was taken to make paraffin sections and frozen sections.Alizarin red staining of paraffin sections was used to detect the formation of calcified nodules in thoracic aorta of mice,and immunofluorescence staining of frozen sections was used to detect the expression of BMP-2 and Runx2 proteins.Results: 1.The percentage of the area of alizarin red stained calcified nodules in the total cross-sectional area of blood vessels was significantly increased after vitamin D3 intervention(p<0.001).Theexpression of Runx2 and BMP-2 protein increased by immunofluorescence staining(p<0.05).2.compared with vitamin D3 intervention group,small dose NBP intervention group,large dose NBP intervention group significantly reduced the percentage of calcified nodule area in total vascular cross-sectional area(p<0.001).The expressions of Runx2 and BMP-2 protein were decreased by immunofluorescence staining(p<0.05).3.compared with the low-dose NBP intervention group and the high-dose NBP intervention group,the percentage of calcified nodule area in the total vascular cross-sectional area was significantly reduced(p<0.05).The expression of Runx2 and BMP-2 protein was decreased by immunofluorescence staining(p<0.05).Conclusion: NBP treatment can inhibit arterial calcification in mice,reduce the expression of BMP-2?Runx2 in calcified blood vessels,and the inhibitory effect of high dose butyphthalide is more significant.