ZBTB20 Positively Regulates Oxidative Stress, Mitochondrial Fission, and Inflammatory Responses of ox‐LDL‐Induced Macrophages in Atherosclerosis

Abstract

Atherosclerosis (AS) is one of the most serious and common cardiovascular diseases affecting human health. AS is featured by the accumulation of plaques in vessel walls. The pathophysiology of AS is relevant in the low-density lipoprotein (LDL) uptake by macrophages, as well as the conversion of macrophages to foam cells. However, the mechanisms about how macrophages regulate AS have not been fully elucidated. In this study, we aimed to illuminate the roles of ZBTB20 and to excavate the underlying regulative mechanisms of ZBTB20 in AS. The microarray analysis revealed that ZBTB20 was a hub gene in the oxidative stress and inflammatory responses induced by oxidized LDL (ox-LDL) in AS. Correspondingly, our validation studies showed that ZBTB20 increased in either the human atherosclerotic lesion or the ox-LDL-stimulated macrophages. Moreover, the knockdown of ZBTB20 decreased M1 polarization, suppressed the proinflammatory factors, inhibited mitochondrial fission, and reduced the oxidative stress level of macrophages induced by ox-LDL. The mechanistic studies revealed that the ZBTB20 knockdown suppressed NF-κB/MAPK activation and attenuated the mitochondrial fission possibly via regulating the nucleus translocation of NRF2, a pivotal transcription factor on redox homeostasis. Our in vivo studies showed that the sh-ZBTB20 adenovirus injection could reduce the progression of AS in apolipoprotein E-deficient (ApoE-/-) mice. All in all, these results suggested that ZBTB20 positively regulated the oxidative stress level, mitochondrial fission, and inflammatory responses of macrophages induced by ox-LDL, and the knockdown of ZBTB20 could attenuate the development of AS in ApoE-/- mice.

BTK Promotes Atherosclerosis by Regulating Oxidative Stress, Mitochondrial Injury, and ER Stress of Macrophages

“…Inflammation and polarization of macrophages play an important role in AS. In our previous report, we also found that atherosclerotic lesions can be alleviated when inflammatory responses of ox-LDL-induced macrophages were released [ 28 ].…”

“…Finally, we had reported that the injection of ZBTB20 knockdown adenovirus in the tail vein of ApoE −/− mice fed with a Western diet could alleviate AS, indicating that the tail vein adenovirus injection is a potential method for treatment of cardiovascular diseases [ 28 ]. In our current study, ApoE −/− mice injected with BTK knockdown adenovirus presented a smaller AS lesion area in the aorta.…”

“…No. YB-002, Yiyuan Biotech) was used at a concentration of 50 μ g/ml as previously described [ 28 , 29 ].…”

“…The mouse AS model was established as described in our previous study [ 28 ]. Briefly, ApoE −/− mice (males, 8 weeks old, and 20 g body weight) with C57BL/6J background were purchased from GemPharmatech Co., Ltd. All mice were reared in a specific pathogen-free environment with a 12 h light/12 h dark cycle at the temperature of 22 ± 2°C and the relative humidity of 40-55%.…”

“…For inner face plaque analyses, the whole aortas without adipose tissue, fixed in 4% paraformaldehyde overnight, were stained with Oil Red O solution for 15 min and washed with 70% ethanol. For aortic root cross-section analysis, the mouse hearts with aortic roots were embedded and sliced into several sections, followed by staining with Oil Red O solution as described in our previous study [ 28 ]. Images were viewed by a biomicroscope (DM200, Leica) [ 47 , 48 ].…”