摘要
目的:研究IL-37对氧化低密度脂蛋白(ox-LDL)诱导的人主动脉血管平滑肌细胞(HA-SMCs)钙化的作用及对模式识别受体(TLR)/核转录因子(NF-κB)通路和成骨转录因子的影响,探讨IL-37在钙化反应中的作用机制。方法:采用oxLDL诱导HA-SMCs钙化模型,模型组加入100 ng/ml IL-37或100μmol/L PDTC预处理,CCK-8检测细胞增殖活性;流式细胞术检测细胞凋亡;茜素红染色检测钙化结节;qPCR检测平滑肌肌动蛋白(SMA)、骨形态发生蛋白(BMP2)、IL-37 mRNA水平;qPCR、Western blot检测TLR、NF-κB、p-NF-κB、RUNX相关转录因子(RUNX2)、碱性磷酸酶(ALP)、SMA、平滑肌22α(SM22α)的mRNA和蛋白表达水平。结果:与对照组相比,ox-LDL处理后细胞增殖活性增强并具有浓度依赖性(P<0.05),凋亡率升高,同时模型组产生明显的橘红色钙化结节,TLR、NF-κB等炎症因子mRNA及蛋白表达上调,IL-37、RUNX2、BMP2、ALP的mRNA表达上调,SMA、SM22α的mRNA表达下降,p-NF-κB、RUNX2的蛋白表达量明显升高;与造模组相比,IL-37预处理组细胞增殖率下降,凋亡率降低(P<0.05),钙化结节明显减少,TLR、NF-κB等炎症因子的mRNA及蛋白表达明显下降,RUNX2、BMP2、ALP的mRNA表达下调,SMA、SM22α的mRNA表达升高,p-NF-κB、RUNX2的蛋白表达量明显下降,与抑制剂PDTC组在细胞因子表达上呈现相同趋势。结论:IL-37可抑制HA-SMCs钙化,可能与抑制NF-κB通路及细胞成骨表型转化相关。
Objective:To investigate the effect of IL-37 on oxidized low-density lipoprotein(ox-LDL)-induced calcification in human aortic vascular smooth muscle cells(HA-SMCs)and the influence of toll-like receptor(TLR)/nuclear transcription factor(NF-κB)pathway and osteogenic transcription factors,to demonstrate the mechanism of IL-37 in calcification.Methods:The calcification model of HA-SMCs was treated with ox-LDL,and the model group was pretreated with 100 ng/ml IL-37 or 100µmol/L PDTC.CCK8 was used to detect cell proliferation activity;flow cytometry was used to detect apoptosis.Alizarin red staining was used to detect calcified nodules.The mRNA levels of smooth muscle actin(SMA),bone morphogenetic protein(BMP2)and IL-37 were detected by qPCR;mRNA and protein expression levels of TLR,NF-κB,p-NF-κB,RUNX-associated transcription factor(RUNX2),alkaline phosphatase(ALP),SMA and smooth muscle 22α(SM22α)were detected by qPCR and Western blot.Results:Compared with the control group,ox-LDL treatment enhanced cellular proliferation activity and was concentration-dependent(P<0.05),and apoptosis rate was increased,while the model group produced significant orange-red calcified nodules,up-regulated mRNA and protein expression of inflammatory factors such as TLR and NF-κB,up-regulated mRNA expression of IL-37,RUNX2,BMP2,ALP,while SMA and SM22αmRNA expressions were decreased,and the protein expressions of p-NF-κB and RUNX2 were increased significantly;compared with the modeling group,the IL-37 pretreatment group showed lower viability and decreased apoptosis rate(P<0.05),significantly reduced calcified nodules,and the mRNA and protein expression of inflammatory factors such as TLR and NF-κB,and mRNA of RUNX2,BMP2,ALP expressions were downregulated,mRNA expression of SMA and SM22αwere elevated,and protein expressions of p-NF-κB and RUNX2 were significantly decreased,showing the same trend as the inhibitor PDTC group in cytokines.Conclusion:IL-37 inhibits calcification of HA-SMCs,which may be related to the inhibition of NF-κB pathway and osteogenic phenotype transformation.
作者
马晨越
贺琼逸
王猛
柴萌
张海涛
MA Chenyue;HE Qiongyi;WANG Meng;CHAI Meng;ZHANG Haitao(Air Force Clinical College,the Fifth Clinical College,Anhui Medical University,Hefei 230032,China;Department of Cardiology,Air Force Special Medical Center,Beijing 100142,China;Hebei North University,Zhangjiakou 075000,China;De-partment of Cardiology,Beijing Anzhen Hospital,Beijing Institute of Cardiopulmonary and Vascular Diseases,Bei-jing Key Laboratory of Precision Medicine for Coronary Atherosclerosis Disease,Clinical Center for Coronary Heart Disease,Capital Medical University,Beijing 100029,China)
出处
《中国免疫学杂志》
北大核心
2025年第2期304-309,共6页
Chinese Journal of Immunology
基金
军队医学科技青年培育计划拔尖项目(20QNPY062)
北京医院管理局孵化项目(PX2022023)。
