Yang Hangzhen, Hong Yimei, Lin Yanya, Li Xin
School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, Guangdong, China.
Department of Emergency Medicine, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Guangzhou 510080, Guangdong, China.
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2022 Jun;34(6):646-650. doi: 10.3760/cma.j.cn121430-20220218-00153.
To investigate the protective effect of nicotinamide phosphoribosyltransferase (NAMPT) on abdominal aortic aneurysm by delaying the senescence of aortic vascular smooth muscle cells (VSMC).
The primary VSMC cells from normal and patients with abdominal aortic aneurysm were cultured by tissue adherence method. Cells were divided into normal human-derived VSMC group (Ctrl-VSMC group), abdominal aortic aneurysm patient-derived VSMC group (AAA-VSMC group), and angiotensin II (Ang II) in vitro abdominal aortic aneurysm model group (Ang II-VSMC group, 100 nmol/L Ang II treated normal human-derived VSMC for 48 hours), Ang II + P7C3 group and AAA + P7C3 group after NAMPT agonist P7C3 intervention (adding 5 μmol/L P7C3 on the basis of Ang II-VSMC group and AAA-VSMC group, respectively). Immunofluorescence staining was used to identify VSMC; cell proliferation-associated antigen Ki67 staining was used to detect cell proliferation; senescence associated β-galactosidase (SA-β-gal) staining was used to detect cell senescence in each group; Western blotting was used to detect the protein expression levels of senescence-related proteins p21, p16 and NAMPT in each group.
Compared with the Ctrl-VSMC group, the positive rate of SA-β-gal staining and the expression levels of senescence-related proteins p21 and p16 in the AAA-VSMC group and Ang II-VSMC group were significantly increased [SA-β-gal staining positive rate: (74.1±4.4)%, (68.6±5.5)% vs. (36.8±10.3)%, p21/GAPDH: 0.61±0.07, 0.51±0.03 vs. 0.31±0.03, p16/GAPDH: 0.77±0.03, 0.72±0.06 vs. 0.33±0.26, all P < 0.01]. However, the expression of NAMPT was significantly decreased (NAMPT/GAPDH: 0.88±0.07, 0.79±0.14 vs. 1.29±0.02, both P < 0.01). Compared with the Ang II-VSMC group, the positive rate of SA-β-gal staining and the expressions levels of senescence-related proteins p21 and p16 in the Ang II + P7C3 group were significantly lower [SA-β-gal staining positive rate: (49.1±3.2)% vs. (68.6±5.5)%, p21/GAPDH: 0.35±0.06 vs. 0.51±0.03, p16/GAPDH: 0.47±0.08 vs. 0.72±0.06, all P < 0.05], while the expression of NAMPT was significantly increased (NAMPT/GAPDH: 1.15±0.06 vs. 0.79±0.14, P < 0.01). Compared with the AAA-VSMC group, the positive rate of SA-β-gal staining and the expression levels of senescence-related proteins p21 and p16 in the AAA+P7C3 group were significantly lower [SA-β-gal staining positive rate: (54.1±6.0)% vs. (74.1±4.4)%, p21/GAPDH: 0.38±0.02 vs. 0.61±0.07, p16/GAPDH: 0.50±0.13 vs. 0.77±0.03, all P < 0.05], but the expression of NAMPT was significantly increased (NAMPT/GAPDH: 1.25±0.28 vs. 0.88±0.07, P < 0.01).
NAMPT agonist P7C3 can delay the senescence of VSMC and play a protective role in abdominal aortic aneurysm.
通过延缓主动脉血管平滑肌细胞（VSMC）衰老，探讨烟酰胺磷酸核糖转移酶（NAMPT）对腹主动脉瘤的保护作用。
采用组织贴壁法培养正常人和腹主动脉瘤患者的原代VSMC细胞。细胞分为正常人源VSMC组（Ctrl-VSMC组）、腹主动脉瘤患者源VSMC组（AAA-VSMC组）、体外腹主动脉瘤模型组（Ang II-VSMC组，用100 nmol/L血管紧张素II（Ang II）处理正常人源VSMC 48小时）、Ang II + P7C3组和AAA + P7C3组（分别在Ang II-VSMC组和AAA-VSMC组基础上加5 μmol/L P7C3）。采用免疫荧光染色鉴定VSMC；采用细胞增殖相关抗原Ki67染色检测细胞增殖；采用衰老相关β-半乳糖苷酶（SA-β-gal）染色检测各组细胞衰老情况；采用蛋白质免疫印迹法检测各组衰老相关蛋白p21、p16和NAMPT的蛋白表达水平。
与Ctrl-VSMC组比较，AAA-VSMC组和Ang II-VSMC组SA-β-gal染色阳性率及衰老相关蛋白p21和p16表达水平显著升高[SA-β-gal染色阳性率：（74.1±4.4）%、（68.6±5.5）%比（36.8±10.3）%，p21/GAPDH：0.61±0.07、0.51±0.03比0.31±0.03，p16/GAPDH：0.77±0.03、0.72±0.06比0.33±0.26，均P < 0.01]。然而，NAMPT表达显著降低（NAMPT/GAPDH：0.88±0.07、0.79±0.14比1.29±0.02，均P < 0.01）。与Ang II-VSMC组比较，Ang II + P7C3组SA-β-gal染色阳性率及衰老相关蛋白p21和p16表达水平显著降低[SA-β-gal染色阳性率：（49.1±3.2）%比（68.6±5.5）%，p21/GAPDH：0.35±0.06比0.51±0.03，p16/GAPDH：0.47±0.08比0.72±0.06，均P < 0.05]，而NAMPT表达显著升高（NAMPT/GAPDH：1.15±0.06比0.79±0.14，P < 0.01）。与AAA-VSMC组比较，AAA+P7C3组SA-β-gal染色阳性率及衰老相关蛋白p21和p16表达水平显著降低[SA-β-gal染色阳性率：（54.1±6.0）%比（74.1±4.4）%，p21/GAPDH：0.38±0.02比0.61±0.07，p16/GAPDH：0.50±0.13比0.77±0.03，均P < 0.05]，但NAMPT表达显著升高（NAMPT/GAPDH：1.25±0.28比0.88±0.07，P < 0.01）。
NAMPT激动剂P7C3可延缓VSMC衰老，对腹主动脉瘤起保护作用。
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