Effect of CD36/Calpain-1/NLRP3 axis on pyroptosis pathway and influence of Th17/Treg inflammatory axis on vascular smooth muscle cells
Wu Lihao
Fu Qiang
Zhan Zhongqun
Liu Hui
Li Yang
Li Jianping
Abstract：Objective To discuss the effect of platelet reactive protein receptor(CD36)/calcium-activated neutral protease(Calpain-1)/NLRP3 inflammasome axis on pyroptosis pathway and influence of T helper 17 cell(Th17)/regulatory T cell(Treg)inflammatory axis on vascular smooth muscle cells(VSMC).Methods Platelet-derived growth factor(PDGF)was used to treat primary VSMC of rats for 24 h,and then a VSMC-induced VSMC mode was established.The treated VSMC were divided into control group,Lv-pcDNA-CD36 group,Lv-si-CD36 group,empty CD36 group,Lv-pcDNA-Calpain-1 group,Lv-si-Calpain-1 group,empty Calpain-1 group,Lv-pcDNA-NLRP3 group,Lv-si-NLRP3 group,and empty NLRP3 group.The expression levels of CD36,Calpain-1 and NLRP3 in VSMC were detected by using qRT-PCR.The proliferation,migration,invasion and pyroptosis of VSMC were detected,respectively,by using MTT,Transwell chamber test and flow cytometer.The expressions of CD36,Calpain-1,NLRP3,Th17,Treg and pyroptosis-related protein(GSDMD)were detected by using Western blotting assay.Results Compared with empty CD36 group,empty Calpain-1 group and empty NLRP3 group,expression levels of CD36 mRNA,Calpain-1 mRNA,NLRP3 mRNA increased in Lv-pcDNA-CD36 group,Lv-pcDNA-Calpain-1 group and Lv-pcDNA-NLRP3 group,and decreased in Lv-si-CD36 group,Lv-si-Calpain-1 group and Lv-si-NLRP3 group(P＜0.05).The differences in expression levels of all observation indexes had no statistical significance among control group,empty CD36 group,empty Calpain-1 group and empty NLRP3 group(P＞0.05).The rate of proliferation,number of migration and invasion and pyroptosis rate,and protein expression levels of CD36,Calpain-1,NLRP3,Th17 and GSDMD increased,and protein expression level of Treg decreased in Lv-pcDNA-CD36 group,Lv-pcDNA-Calpain-1 group and Lv-pcDNA-NLRP3 group compared with control group,empty CD36 group,empty Calpain-1 group and empty NLRP3 group(P＜0.05).The rate of proliferation,number of migration and invasion and pyroptosis rate,and protein expression levels of CD36,Calpain-1,NLRP3,Th17 and GSDMD decreased,and protein expression level of Treg increased in Lv-si-CD36 group,Lv-si-Calpain-1 group and Lv-si-NLRP3 group compared with Lv-pcDNA-CD36 group,Lv-pcDNA-Calpain-1 group and Lv-pcDNA-NLRP3 group(P＜0.05).Conclusion When CD36/Calpain-1/NLRP3 pathway is activated,the dysregulation of Th17/Treg inflammatory axis,and further VSMC pyroptosis can be induced.
Keywords：Platelet reactive protein receptorCalcium-activated neutral proteasePyroptosisSmooth muscle cellsT helper 17 cellRegulatory T cellRats
Publication Date：2025-04-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：8( 437-444 )