Regulation of Ca2+/NLRP3/Caspase-1 signaling pathway in atherosclerotic endothelial dysfunction
Objective To explore the regulation of Ca2+/NLRP3/Caspase-1 signaling pathway on endothelial dysfunction in atherosclerosis(AS).Methods 16 male ApoE-/-mice were randomly divided into a normal diet group and a high-fat diet group and measured for the weight on an empty stomach every week;The mice were fed continuously for 20 weeks and then sacrificed,of which the arterial tissues were collected;Serum cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL-C)and high density lipoprotein(HDL-C)were detected by biochemical analyzer;The thickness of aortic luminal plaque and intima was assessed by oil red O staining.Human umbilical vein endothelial fusion cells EA.hy926 were divided into control group,high-sugar and high-fat group,and Ca2+inhibitor group,which were detected for Ca2+content by flow cytometry at different time periods,for the expression of NLRP3 by immunofluorescence,and for the protein expression levels of NLRP3,Caspase-1/p10/p20,VCAM-1 and ICAM-1 in aortic tissue and cells by Western blot.Results Compared with the normal diet group,the weight of the mice in high-fat diet group increased significantly(F=3.333,P=0.026),the serum TC,TG and LDL-C significantly increased,while HDL-C significantly decreased(F=1.852,2.410,1.920 and 2.917,P=0.000,0.004,0.002 and 0.003,respectively);The aortic plaque surface area/aortic surface area increased significantly(F=3.256,P=0.000),plaque cross-sectional area/aortic lumen area increased significantly(F=6.433,P=0.008),and NLRP3,Caspase-1/p10/p20,VCAM-1 and ICAM-1 protein expressions increased significantly(F=8.997,4.664,4.486 and 9.949,P=0.036,0.022,0.005 and 0.018,respectively).Compared with the control group,the Ca2+content of endothelial cells in high-sugar and high-fat group increased in the 6 h,12 h,and 24 h(F=0.310,5.649 and 5.580,P=0.006,0.009 and 0.004,respectively),especially in the 6 h group;The expression of NLRP3 in high-sugar and high-fat group increased significantly;VCAM-1,ICAM-1 and Caspase-1/p10/p20 protein expression increased significantly(F=10.476,5.310 and 9.306,P=0.029,0.030 and 0.018,respectively).Compared with the high-sugar and high-fat group,the expression of NLRP3 and Caspase1/p10/p20 protein in EA.hy926 cells of Ca2+inhibitor group decreased significantly(F=2.196,2.882 and 0.035,P＜0.01,＜0.05 and＜0.05,respectively).Conclusion NLRP3 inflammasome induced dysfunction of vascular endothelial cells and aggravated the progression of AS,which might be closely related to Ca2+and downstream signaling pathways.
Vascular endothelial injuryAtherosclerosisNLRP3 inflammasomeHigh sugar and high fatCa2+
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