引用本文:魏春阳,齐国先,王衍富,张华,张多多.TWEAK及Fn14在衰老大鼠血管平滑肌细胞中的表达及意义[J].大连医科大学学报,2021,43(3):198-203,223.
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TWEAK及Fn14在衰老大鼠血管平滑肌细胞中的表达及意义
魏春阳1,2, 齐国先1, 王衍富2, 张华2, 张多多3
1.中国医科大学附属第一医院 老年医学科, 辽宁 沈阳 110001;2.大连医科大学附属第一医院 老年医学科, 辽宁 大连 116011;3.大连医科大学附属第一医院 心律失常科, 辽宁 大连 116011
摘要:
目的 探讨肿瘤坏死因子样弱凋亡诱导因子(tumor necrosis factor-like weak inducer of apoptosis,TWEAK)及其受体成纤维细胞生长因子诱导蛋白14(fibroblast growth factor-inducing protein 14,Fn14)对血管平滑肌细胞衰老的潜在作用。方法 将大鼠主动脉平滑肌(A10)细胞分为对照组及AngⅡ组,经同步化后,对照组10%血清培养基培养,AngⅡ组加1×10-7 mol/L的AngⅡ培养。再将A10细胞分为对照组、未感染组、空载体组及TWEAK过表达组,经同步化后,对照组、未感染组不做转染,空载体组及TWEAK过表达组分别转染空载体腺病毒及TWEAK过表达腺病毒。转染48 h后,未感染组、空载体组及TWEAK过表达组加1×10-7 mol/L的AngⅡ培养,对照组10%血清培养基培养。通过衰老相关β-半乳糖苷酶(SA-β-Gal)染色、Western blot检测P53、P21来评估细胞衰老情况,实时荧光定量PCR、Western blot检测TWEAK mRNA及TWEAK、Fn14蛋白表达水平。结果 随着AngⅡ作用时间延长,AngⅡ组SA-β-Gal染色逐渐增多,P53、P21、TWEAK、Fn14蛋白表达明显增多,与对照组比较,差异均有统计学意义(P<0.05)。TWEAK过表达组血管平滑肌细胞,在AngⅡ诱导衰老时Fn14、P53、P21的表达显著增多,与对照组、未感染组、空载体组比较,差异均有统计学意义(P<0.05)。结论 血管平滑肌细胞衰老时TWEAK、Fn14表达增多,TWEAK对血管平滑肌细胞衰老有促进作用。
关键词:  TWEAK  Fn14  血管老化  血管平滑肌细胞  衰老
DOI:10.11724/jdmu.2021.03.02
分类号:Q255
基金项目:
Significance of TWEAK expression in rat vascular smooth muscle cell senescence
WEI Chunyang1,2, QI Guoxian1, WANG Yanfu2, ZHANG Hua2, ZHANG Duoduo3
1.Department of Gerontology, the First Affiliated Hospital of China Medical University, Shenyang 110001, China;2.Department of Gerontology, the First Affiliated Hospital of Dalian Medical University, Dalian 116011, China;3.Department of Arrhythmia, the First Affiliated Hospital of Dalian Medical University, Dalian 116011, China
Abstract:
Objective To investigate the potential effects of tumor necrosis factor-like weak inducer of apoptosis (TWEAK)and its receptor fibroblast growth factor-inducing protein 14 (Fn14) on vascular smooth muscle cell senescence. Methods First,the rat aortic smooth muscle (A10) cells were divided into control group and Ang Ⅱ group. After synchronization, the control group was cultured in 10% serum medium, and Ang Ⅱ group was added with Ang Ⅱ (1×10-7 mol/L). Then,A10 cells were divided into control group, uninfected group, empty vector group and tweak overexpression group. After synchronization, control group and uninfected group were not transfected, empty vector group and tweak overexpression group were transfected with empty vector adenovirus and tweak overexpression adenovirus respectively. After 48 hours of transfection, the uninfected group, empty vector group and tweak overexpression group were added with Ang Ⅱ, and the control group was cultured in 10% serum medium. SA-β-Gal staining was used to show the level of cell senescence. The senescence markers, P53, P21, were analyzed by Western blot.TWEAK and Fn14 expression were examined by Real-time quantitative PCR and Western blot assays. Results In Ang Ⅱ group, SA-β-Gal positive cells increased significantly. P53, P21, TWEAK, Fn14 expression were significantly increased than those in the control group (P<0.05). The expression of Fn14, P53, P21 in TWEAK overexpression group were higher significantly than those in the control group, uninfected group, and empty vector group (P<0.05). Conclusion TWEAK could promote aging of vascular smooth muscle cells.
Key words:  TWEAK  Fn14  blood vessel aging  vascular smooth muscle cell  senescence