Palmitate induces VSMC apoptosis via toll like receptor (TLR)4/ROS/p53 pathway. (August 2017)

• Record Type: Journal Article
• Authors: Zhang, Yuanjun; Xia, Guanghao; Zhang, Yaqiong; Liu, Juxiang; Liu, Xiaowei; Li, Weihua; Lv, Yaya; Wei, Suhong; Liu, Jing; Quan, Jinxing
• Is Part Of: Atherosclerosis. Volume 263(2017)
• Page Start: 74
• Page End: 81
• Publication Date: 2017-08
• Subjects: Palmitate -- Vascular smooth muscle cells -- Apoptosis -- Oxidative stress -- Toll-like receptor 4; Arteriosclerosis -- Periodicals; Electronic journals; 616.136
• DOI: 10.1016/j.atherosclerosis.2017.06.002
• Languages: English
• ISSNs: 0021-9150

Abstract

Abstract: Background and aims: Toll-like receptor 4 ( TLR4 ) has been implicated in vascular inflammation, as well as in the pathogenesis of atherosclerosis and diabetes. Vascular smooth muscle cell (VSMC) apoptosis has been shown to induce plaque vulnerability in atherosclerosis. Previous studies reported that palmitate induced apoptosis in VSMCs; however, the role of TLR4 in palmitate-induced apoptosis in VSMCs has not yet been defined. In this study, we investigated whether or not palmitate-induced apoptosis depended on the activation of the TLR4 pathway. Methods: VSMCs were treated with or without palmitate, CRISPR/Cas9z-mediated genome editing methods were used to deplete TLR4 expression, while NADPH oxidase inhibitors were used to inhibit reactive oxygen species (ROS) generation. Cell apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, ROS was measured using the 2′, 7'-dichlorodihydrofluorescein diacetate (DCFH-DA) method, the mRNA and protein expression levels of caspase 3, caspase 9, BCL-2 and p53 were studied by real-time polymerase chain reaction (RT-PCR) and ELISA. Results: Palmitate significantly promotes VSMC apoptosis, ROS generation, and expression of caspase 3, caspase 9 and p53 ; while NADPH oxidase inhibitor pretreatment markedly attenuated these effects. Moreover, knockdown of TLR4 significantly blocked palmitate-induced ROS generation and VSMC apoptosis accompanied by inhibition of caspase 3, caspase 9, p53 expression and restoration of BCL-2 expression. Conclusions: Our results suggest that palmitate-induced apoptosis depends on the activation of the TLR4 /ROS/ p53 signaling pathway, and that TLR4 may be a potential therapeutic target for the prevention and treatment of atherosclerosis. Highlights: VSMCs apoptosis is associated with plaque vulnerability in atherosclerosis. Palmitate induces apoptosis through ROS generation and p53 activation in VSMCs. TLR4 knockdown blocked ROS generation, p53 activation and apoptosis induced by palmitate in VSMCs. TLR4 may be a potential therapeutic target for the prevention and treatment of atherosclerosis.