基于NF-κB/NLRP3/Caspase-1通路探讨养心舒脉颗粒对血管紧张素Ⅱ诱导动脉内皮细胞焦亡及动脉粥样硬化斑块的影响OA

Objective It is to explore the effect of Yang-Xin-Shu-Mai Granule(YXSMG)on angiotensin Ⅱ induced arterial endothelial cell pyroptosis and atherosclerosis plaque via regulating nuclear factor-κB(NF-κB)/Nod-like receptor protein 3(NLRP3)/Caspase-1 pathway.Methods Six normal C57BL/6J male mice(used as blank group)and18 ApoE-/-mice with the same genetic background were selected for the experiment.The 18 ApoE-/-mice were randomly divided into model group,low-dose and high-dose groups of YXSMG,with 6 mice in each group.The mice of all the four groups were fed with high-fat diet,and the ApoE-/-mice were implanted subcutaneously with Alzet osmotic pump to treated with angio-tensin Ⅱ 500 ng/(kg·min)for4 weeks to establish models of atherosclerosis.The low-dose and high-dose groups of YX-SMG were respectively treated with YXSMG aqueous solution at 5.2 g/(kg·d)and 11.7 g/(kg·d)via gavage,the blank group and model group were gavaged with the same volume of sterile distilled water,all once daily,continuously trea-ted for 4 weeks.The changes in body weight and blood pressure of mice in each group during experiment were observed,the mice were sacrificed at the end of treatment,their aorta histopathology was observed by HE and Movat staining;the positive expressions of NLRP3,phosphorylated NF-κB p65(p-NF-κB p65)and gasdermin D(GSDMD)in aortic sinuses were detected by immunohistochemistry;the protein expressions of NLRP3,p-NF-κB p65,NF-κB p65,cleaved Caspase-1,Caspase-1,and GSDMD in aortic plaques were detected by Western blot.Results After modeling,the body weight,sys-tolic blood pressure and diastolic blood pressure of mice in the model group,low-dose,and high-dose groups of YXSMG were all significantly higher than those of mice in the control group at the same time point(all P<0.05),but there were no statistically significant differences in body weight,systolic blood pressure or diastolic blood pressure among the model group,low-dose,and high-dose groups of YXSMG at the same time point(all P>0.05).Compared with the blank group,there were much more aortic atherosclerosis plaque gathered around with inflammatory cells,and much more endothelial cell nuclei,elastic fibers,collagen fibers,and pale purple foam cells in aortic sinus in mice of the model group;strong positive expressions of NLRP3,p-NF-κB p65 and GSDMD were observed in the aortic sinuses,and the relative protein expressions of NLRP3,p-NF-κB p65/NF-κB p65,cleaved Caspase-1/Caspase-1 and GSDMD in aortic plaques were all significantly increased(all P<0.05).Compared with the model group,the aortic plaques with inflammatory cell infiltration,pale pur-ple foam cells beneath the endothelial cells of the aortic sinus were decreased,along with degeneration and thinning of elas-tic fibers in mice of the low-dose,and high-dose groups of YXSMG;the positive expressions of NLRP3,p-NF-κB p65 and GSDMD in aortic sinuses,and the relative protein expressions of NLRP3,p-NF-κB p65/NF-κB p65,cleaved Caspase-1/Caspase-1,and GSDMD in aortic plaques(except for the relative expression of p-NF-κB p65/NF-κB p65 in the low-dose group)were all significantly reduced(all P<0.05).Conclusion YXSMG can delay the progression of atherosclerosis,and its mechanism may be related to regulating the NF-κB/NLRP3/Caspase-1 pathway to inhibit the pyroptosis of arterial endothelial cells.