Protein disulfide-isomerase A3 knockdown attenuates oxidized low-density lipoprotein-induced oxidative stress, inflammation and endothelial dysfunction in human umbilical vein endothelial cells by downregulating activating transcription factor 2

Abstract

Atherosclerosis is a chronic inflammatory disease implicated in oxidative stress and endothelial dysfunction. Protein disulfide-isomerase A3 (PDIA3) has been reported to regulate oxidative stress and suppress inflammation. This study aimed to explore the function of PDIA3 in atherosclerosis and the underlying mechanisms. PDIA3 expression in oxidized low-density lipoprotein (ox-LDL)-induced human umbilical vein endothelial cells (HUVECs) was detected using RT-qPCR and Western blotting. Following PDIA3 knockdown through transfection with small interfering RNA targeting PDIA3, cell viability, oxidative stress and inflammation in ox-LDL-induced HUVECs was examined using a Cell Counting Kit-8, corresponding kits and ELISA, respectively. The levels of CD31, α-smooth muscle, iNOS, p-eNOS, eNOS and NO were assessed using RT-qPCR, Western blotting and an NO kit to reflect endothelial dysfunction in ox-LDL-induced HUVECs. The relationship between PDIA3 and the activating transcription factor 2 (ATF2) was confirmed using co-immunoprecipitation. In addition, ATF2 expression was examined following PDIA3 silencing. The results indicated that PDIA3 was highly expressed in ox-LDL-induced HUVECs. PDIA3 silencing increased cell viability, and reduced oxidative stress and inflammation, as evidenced by the decreased levels of reactive oxygen species, malondialdehyde, TNF-α, IL-1β and IL-6, and increased superoxide dismutase and glutathione peroxidase activity. In addition, PDIA3 deletion improved endothelial dysfunction. PDIA3 interacted with ATF2, and PDIA3 deletion downregulated ATF2 expression. Furthermore, ATF2 overexpression reversed the effects of PDIA3 knockdown on ox-LDL-induced damage of HUVECs. Collectively, PDIA3 knockdown was found to attenuate ox-LDL-induced oxidative stress, inflammation and endothelial dysfunction in HUVECs by downregulating ATF2 expression, showing promise for the future treatment of atherosclerosis.

Expression and Prognostic Significance of PDIA3 in Cervical Cancer

“…Under normal circumstances, the major part of PDIA3 is associated with the ER and forms complex with calreticulin and calnexin to participate in the correct folding and quality control of new synthetic glycoproteins secreted or localized to cell membranes, which is not only capable of protein modification and folding but also capable of exerting the effects of group thinning and oxidation reduction [6]. PDIA3 is expressed in various types of human cancers, including ovarian cancer, breast cancer, uterine cancer, lung cancer, gastric cancer, and hepatocellular carcinoma [7], and its expression is related to the prognosis and survival of various cancers [8,9]. For example, in patients with diffuse glioma, clear cell renal cell carcinoma, and hepatocellular carcinoma, high expression of PDIA3 is associated with poor survival outcomes [10].…”

Bioinformatics-based identification of lipid- and immune-related biomarkers in abdominal aortic aneurysms

“…To date, no studies have reported the role of PDIA3, TYROBP, HSPA1A and HCK and in onset and development of AAA, especially in lipid metabolism and the immune system. In atherosclerosis similar to AAA, knockdown of PDIA3 was downregulate ATF2 expression, thereby alleviating OX-LDL-induced oxidative stress, inflammation and endothelial dysfunction in HUVECs [ 44 ]. Pravastatin reduces the expression of TYROBP, which facilitates the reduction of plaque area, lipid deposits and inflammatory response in atherosclerosis [ 45 ].…”

Using integrated GC-MS analysis, in vitro experiments, network pharmacology: exploring migao fatty oil active components/mechanisms against coronary heart disease

“…Oxidized low-density lipoprotein is involved in the production of adhesion molecules and the recruitment of monocytes to the subendothelial region, affecting oxidative stress, inflammation, and endothelial dysfunction. Monocytes multiply and develop into macrophages, hastening plaque formation, local inflammation, and thrombosis (Jia et al, 2022;Xiong et al, 2022). Therefore, for prevention, lipid aggregation and oxidation must be avoided.…”