摘要
目的探讨ABI家族成员3结合蛋白(ABI family member 3-binding protein,ABI3BP)在血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导内皮祖细胞功能障碍中的作用及机制。方法为探讨ABI3BP在AngⅡ诱导内皮祖细胞功能障碍中的作用,将细胞分为4组,sh-NC组[转染阴性对照短发夹RNA(LV-scramble-shRNA)+磷酸盐缓冲液(phosphate buffered saline,PBS)]、sh-ABI3BP组[转染ABI3BP shRNA(LV-ABI3BP-shRNA)+PBS]、sh-NC+AngⅡ组(LV-scramble-shRNA+AngⅡ)和sh-ABI3BP+AngⅡ组(LV-ABI3BP-shRNA+AngⅡ)。采用Transwell实验检测细胞迁移能力,黏附实验检测细胞黏附能力,Matrigel检测细胞成管能力,原位末端标记法检测细胞凋亡。Western blot检测整合素β1-黏着斑激酶(focal adhesion kinase,FAK)-P53信号通路变化情况。结果与sh-NC组比较,sh-NC+AngⅡ组迁移细胞数量、黏附细胞数量、小管形成数量显著降低,细胞凋亡率、整合素β1、磷酸化FAK(p-FAK)/FAK及P53蛋白表达显著增高,差异有统计学意义(P<0.05)。与sh-NC+AngⅡ组比较,sh-ABI3BP+AngⅡ组迁移细胞数量[(88.67±8.33)个vs(62.33±7.37)个]、黏附细胞数量[(104.33±6.03)个vs(68.33±10.05)个]、小管形成数量[(36.33±3.21)个vs(19.33±3.06)个]显著增高,细胞凋亡率、整合素β1、p-FAK/FAK及P53蛋白表达水平显著降低,差异有统计学意义(P<0.05)。结论AngⅡ可上调ABI3BP表达,敲低ABI3BP基因表达可改善AngⅡ诱导的内皮祖细胞功能障碍,其机制可能与抑制整合素β1-FAK-P53信号通路有关。
Objective To investigate the role and mechanism of ABI family member 3-binding protein(ABI3BP)in dysfunction of endothelial progenitor cells(EPC)induced by angiotensinⅡ(AngⅡ).Methods EPC were divided into sh-RNA negative control(sh-NC)group(transfected with LV-scramble-shRNA+PBS),sh-ABI3BP group(transfected with LV-ABI3BP-shRNA+PBS),sh-NC+AngⅡgroup(transfected with LV-scramble-shRNA+AngⅡ)and sh-ABI3BP+AngⅡgroup(transfected with LV-ABI3BP-shRNA+AngⅡ)to investigate the effect of silencing ABI3BP on the dysfunction of EPC induced by AngⅡ.Transwell assay,adhesion assay,Matrigel tube formation assay,and TUNEL staining were performed respectively to detect the migration,adhesion and tube formation abilities and cell apoptosis in above cell groups.The expression changes in integrin-β1/FAK/P53 signaling pathway were detected by Western blotting.Results Compared to the sh-NC group,the sh-NC+AngⅡgroup showed significant decreases in the numbers of migratory cells,adhesion cells,and tubule formation,along with increases in the apoptotic rate and the expression levels of Integrinβ1,p-FAK,and P53(P<0.05).While,the sh-ABI3BP+AngⅡgroup had obviously more migratory cells(88.67±8.33 vs 62.33±7.37 units,P<0.05),adhesion cells(104.33±6.03 vs 68.33±10.05 units,P<0.05),and tubule formation(36.33±3.21 vs 19.33±3.06 units,P<0.05),while decreased apoptotic rate and expression levels of integrin-β1,p-FAK and P53 protein when compared with the sh-NC+AngⅡgroup(P<0.05).Conclusion AngⅡcan up-regulate the expression of ABI3BP,and knockdown of ABI3BP can improve AngⅡ-induced EPC dysfunction,which may be related to its inhibition on integrin-β1/FAK/P53 signaling pathway.
出处
《中华老年心脑血管病杂志》
CAS
北大核心
2024年第8期948-953,共6页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基金
湖北省自然科学基金项目(2022CFA036,2024AFB363)。
