摘要
目的探讨swiprosin-1在动脉粥样硬化组织中的表达及对巨噬细胞凋亡和炎症因子表达的影响。方法构建动脉粥样硬化小鼠模型,RT-PCR和Western blot检测动脉粥样硬化组织中swiprosin-1的表达水平。用氧化型低密度脂蛋白（ox-LDL）处理巨噬细胞THP-1、平滑肌细胞VSMC、内皮细胞EC-304,RT-PCR和Western blot检测swiprosin-1的表达水平。巨噬细胞THP-1转染swiprosin-1 siRNA和siRNA control,经ox-LDL处理后,流式细胞术检测细胞凋亡,Western blot检测cleaved Caspase-3、cleaved Caspase-9表达,ELISA检测培养液上清中肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）含量。结果 swiprosin-1在动脉粥样硬化组织中的表达水平明显高于正常组织。swiprosin-1在ox-LDL处理后的巨噬细胞THP-1中表达水平最高,而在平滑肌细胞VSMC、内皮细胞EC-304中表达水平极低。ox-LDL处理后巨噬细胞THP-1凋亡率高达21.64%±1.88%,细胞中cleaved Caspase-3、cleaved Caspase-9表达水平升高,细胞培养液上清中TNF-α、IL-6含量也明显升高。干扰swiprosin-1表达后的巨噬细胞THP-1经ox-LDL处理后凋亡率下降为15.25%±1.10%,细胞中cleaved Caspase-3、cleaved Caspase-9表达水平降低,细胞培养液上清中TNF-α、IL-6含量也明显下降。结论 swiprosin-1在动脉粥样硬化组织中表达升高,降低swiprosin-1表达能够抑制ox-LDL诱导的巨噬细胞凋亡和巨噬细胞炎症因子分泌。
Aim To investigate the expression of swiprosin-1 in atherosclerotic tissue and its influence on apoptosis and inflammatory factors expression in macrophages. Methods Establishment of atherosclerosis mouse model,RT-PCR and Western blot were used to detect the expression of swiprosin-1 in atherosclerotic tissues. Treatment of macrophage THP-1,smooth muscle cell VSMC,endothelial cell EC-304 with ox-LDL,the expression levels of swiprosin-1 were detected by RT-PCR and Western blot. Macrophage THP-1 was transfected with swiprosin-1 siRNA,and siRNA control,after ox-LDL treatment,apoptosis was detected by flow cytometry,the expression of cleaved Caspase-3 and cleaved Caspase-9 were detected by Western blot,TNF-α and IL-6 in the supernatant of culture medium were detected by ELISA.Results The expression level of swiprosin-1 in atherosclerotic tissue was higher than that in normal tissue. The expression level of swiprosin-1 in macrophage THP-1 after ox-LDL treatment was the highest,in smooth muscle cell VSMC and endothelial cell EC-304 were very low. The apoptotic rate of macrophage THP-1 treated with ox-LDL was as high as 21.64% ±1.88%,the expression levels of cleaved Caspase-3 and cleaved Caspase-9 in cells were increased,the levels of TNF-α and IL-6 in the supernatant of culture medium were also significantly increased. The apoptotic rate of macrophage THP-1 treated by ox-LDL after swiprosin-1 interference decreased to 15. 25% ± 1. 10%,the expression levels of cleaved Caspase-3 and cleaved Caspase-9 in cells were decreased,the contents of TNF-α and IL-6 in the supernatant of culture medium also decreased obviously. Conclusion Elevated expression of swiprosin-1 in atherosclerotic tissue,reducing swiprosin-1 expression could inhibit apoptosis and inflammatory cytokine secretion in macrophages induced by ox-LDL.
作者
秦学鹏
陈小龙
王小雪
QIN Xue-Peng;CHEN Xiao-Long;WANG Xiao-Xue(Cardiovascular Medicine Department II Ward, the First Affiliated Hospital of Nanyang Medical College;Department of Nephrology , Nanyang First People＇s Hospital, Nanyang , Henan 473000, China)
出处
《中国动脉硬化杂志》
CAS
2018年第5期479-484,共6页
Chinese Journal of Arteriosclerosis
