ROLE OF PYROPTOSIS OF AORTIC VALVE INTERSTITIAL CELLS IN CALCIFIC AORTIC VALVE DISEASE

Abstract

Objective To culture aortic valve interstitial cells (AVICs) in vitro and construct a calcification model, and to explore the role of pyroptosis in calcific aortic valve disease. Methods We used a calcification induction medium to induce calcification of AVICs and then divided them into control group (standard medium culture), calcification group (calcification induction medium culture), and inhibitor group (24 h VX-765 pretreatment followed by calcification induction medium culture) according to the treatment. CCK-8 assay was used to measure the cell viability of AVICs in each group. Alizarin red staining was used to mea-sure the expression levels of calcium nodules in the AVICs of each group. Western blotting was used to measure the relative expression levels of calcification-related proteins OPN and RunX2, pyroptosis-related proteins NLRP3, GSDMD, Caspase-1, GSDMD-N, and Cleaved-Caspase-1, and inflammatory factors interleukin (IL)-1β and IL-18. Results Compared with the control group, the calcification group showed significant increases in calcium nodules in AVICs and the relative expression levels of calcification-related proteins OPN and RunX2 and pyroptosis-related proteins NLRP3, GSDMD, Caspase-1, GSDMD-N, and Cleaved-Caspase-1 (t=4.586-15.842,P<0.01). Compared with the calcification group, the inhibitor group showed significant reductions in calcium no-dules in AVICs and the relative expression levels of calcification-related proteins OPN and RunX2, pyroptosis-related proteins NLRP3, GSDMD, Caspase-1, GSDMD-N, and Cleaved-Caspase-1, and inflammatory factors IL-1β and IL-18 (t=5.789-19.227,P<0.01). Conclusion There is pyroptosis in the calcification process of AVICs, and VX-765 pretreatment before calcification induction can inhibit the pyroptosis of AVICs to a certain extent.

Keywords

• calcific aortic valve disease|aortic valve interstitial cells|pyroptosis|inflammatory|vx-765