KLF11 protects against abdominal aortic aneurysm through inhibition of endothelial cell dysfunction
Guizhen Zhao, Ziyi Chang, Yang Zhao, Yanhong Guo, Haocheng Lu, Wenying Liang, Oren Rom, Huilun Wang, Jinjian Sun, Tianqing Zhu, Yanbo Fan, Lin Chang, Bo Yang, Minerva T. Garcia-Barrio, Y. Eugene Chen, Jifeng Zhang
Guizhen Zhao, Ziyi Chang, Yang Zhao, Yanhong Guo, Haocheng Lu, Wenying Liang, Oren Rom, Huilun Wang, Jinjian Sun, Tianqing Zhu, Yanbo Fan, Lin Chang, Bo Yang, Minerva T. Garcia-Barrio, Y. Eugene Chen, Jifeng Zhang
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Research Article Vascular biology

KLF11 protects against abdominal aortic aneurysm through inhibition of endothelial cell dysfunction

Abstract

Abdominal aortic aneurysm (AAA) is a life-threatening degenerative vascular disease. Endothelial cell (EC) dysfunction is implicated in AAA. Our group recently demonstrated that Krüppel-like factor 11 (KLF11) plays an essential role in maintaining vascular homeostasis, at least partially through inhibition of EC inflammatory activation. However, the functions of endothelial KLF11 in AAA remain unknown. Here we found that endothelial KLF11 expression was reduced in the ECs from human aneurysms and was time dependently decreased in the aneurysmal endothelium from both elastase- and Pcsk9/AngII-induced AAA mouse models. KLF11 deficiency in ECs markedly aggravated AAA formation, whereas EC-selective KLF11 overexpression markedly inhibited AAA formation. Mechanistically, KLF11 not only inhibited the EC inflammatory response but also diminished MMP9 expression and activity and reduced NADPH oxidase 2–mediated production of reactive oxygen species in ECs. In addition, KLF11-deficient ECs induced smooth muscle cell dedifferentiation and apoptosis. Overall, we established endothelial KLF11 as a potentially novel factor protecting against AAA and a potential target for intervention in aortic aneurysms.

Authors

Guizhen Zhao, Ziyi Chang, Yang Zhao, Yanhong Guo, Haocheng Lu, Wenying Liang, Oren Rom, Huilun Wang, Jinjian Sun, Tianqing Zhu, Yanbo Fan, Lin Chang, Bo Yang, Minerva T. Garcia-Barrio, Y. Eugene Chen, Jifeng Zhang

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Figure 5

KLF11 attenuates ROS production through NOX2 suppression.

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KLF11 attenuates ROS production through NOX2 suppression. (A and B) The ...
(A and B) The production of superoxide was determined in HAECs. HAECs were infected with Ad-lacZ or Ad-KLF11, or Ad-shlacZ or Ad-shKLF11 (10 MOI). After 48 hours, they were treated with TNF-α (2 ng/mL) or AngII (1 μM) for 2 hours in the presence of superoxide detection solution (fluorescent probes). (C–E) HAECs were infected with Ad-GFP or Ad-KLF11 (10 MOI) or transfected with control siRNA (siControl) or KLF11 siRNA (siKLF11, 20 μM). After 48 hours, they were stimulated with TNF-α (2 ng/mL) or AngII (1 μM) for 24 hours. (C and D) The mRNA levels of NOX1, NOX2, NOX4, and NOX5 were determined by qPCR. (E) Western blot to determine the expression of NOX2 in HAECs. (F) HAECs were transfected with siControl, siKLF11 or siKLF11+NOX2 siRNA (siNOX2) (20 μM). After 48 hours, they were treated with TNF-α (2 ng/mL) or AngII (1 μM) for 2 hours in the presence of superoxide detection solution. (G and H) Representative DHE staining and quantification of superoxide production in HAECs. HAECs were transfected with siControl, siKLF11, or siKLF11+siNOX2. After 48 hours, they were pretreated with NOX2 inhibitor (NOX2i, GSK2795039, 1 μM) for 1 hour, then stimulated with TNF-α (2 ng/mL) or AngII (1 μM) for 2 hours, followed by DHE staining of superoxide (red) and immunofluorescence staining of VE-cadherin (green). Nuclei stained by DAPI are shown in blue. Scale bar: 20 μm. (I) HAECs were infected with Ad-lacZ or Ad-flag-KLF11. After 48 hours, they were stimulated with TNF-α (2 ng/mL) for 4 hours, followed by ChIP assay using an antibody against flag or IgG. Data are presented as mean ± SEM. Two-way ANOVA followed by Holm-Sidak post hoc analysis (A–D, F, H, and I).