An Egr-1-specific DNAzyme regulates Egr-1 and proliferating cell nuclear antigen expression in rat vascular smooth muscle cells

Abstract

The aim of the present study was to transfect rat aortic smooth muscle cells with an early growth response factor-1 (Egr-1)-specific DNAzyme (ED5), to observe its effect on Egr-1 and proliferating cell nuclear antigen (PCNA) expression and to elucidate the mechanism of ED5-mediated inhibition of vascular smooth muscle cell (VSMC) proliferation. VSMCs in primary culture obtained by tissue block adhesion were identified by morphological observation and α smooth muscle actin (α-SM-actin) immunocytochemistry. The cells were then transfected with ED5 or scrambled ED5 (ED5SCR). The three groups of cells used in the present study were the control group, ED5 group and ED5SCR group. The expression levels of Egr-1 and PCNA protein were detected following transfection by analyzing and calculating the integral optical density value in each group. Primary culture of VSMCs and transfection of ED5 and ED5SCR were successfully accomplished. Following stimulation with 10% fetal calf serum, the Egr-1 protein was expressed most strongly at 1 h and demonstrated a declining trend over time; the expression of PCNA protein began at 4 h, peaked at 24 h and then demonstrated a slightly declining trend over time. Compared with the control group and the ED5SCR group, ED5 inhibited the expression of Egr-1 and PCNA (P<0.05). ED5 was able to inhibit the expression of Egr-1 and PCNA proteins in VSMCs to a certain extent and VSMC proliferation in vitro. DNAzyme gene therapy may be useful as a new method for treating vascular proliferative diseases, including atherosclerosis and restenosis.

Effects of Tilianin on Proliferation, Migration and TGF-β/Smad Signaling in Rat Vascular Smooth Muscle Cells Induced with Angiotensin II

“…A variety of etiological factors can contribute to blood vessels injury and promote VSMC proliferation and secretion of cytokines. These factors can regulate cell adhesion on the surface of blood vessels, control the synthesis and degradation of extracellular matrix and eventually lead to the development of fiber plaques that are characteristic of AS (Zhang et al , ).…”

Knockdown of connexin 43 attenuates balloon injury-induced vascular restenosis through the inhibition of the proliferation and migration of vascular smooth muscle cells

“…Cell culture. The culture of the VMSCs was performed as previously described (21). Briefly, the rats were anesthetized with an intraperitoneal injection of Hydral (10%, 3.5 ml/kg; Harbin Pharmaceutical Group Co., Ltd., Harbin, China) and the carotid artery was quickly removed under aseptic conditions.…”

DNA enzymes as potential therapeutics: towards clinical application of 10-23 DNAzymes

“…Most recent studies have demonstrated that an Egr-1-specific Dz (ED5) inhibits the expression of Egr-1 and proliferating cell nuclear antigen (PCNA) in vascular SMCs (VSMCs) and VSMC proliferation in vitro [125]. Moreover, ED5 Dz was able to prevent stenosis and occlusion of autogenous vein graft in rats due to the reduction of Egr-1 expression in autogenous vein grafts, effectively restraining VSMC proliferation and intimal hyperplasia [126].…”

“…A key mediator of oxidative stress by affecting apoptosis signal-regulating kinase 1 and MAPK kinase activities; its expression is stimulated in heart tissue after acute myocardial infarction [120] Early growth response-1 A zinc finger transcription factor upregulated by mechanical injury to vascular SMCs and endothelial cells [121][122][123]125,126] c-jun The protein is involved in regulating proliferation, transformation and apoptosis; it contains 'leucine zipper' domain and forms homo-and heterodimers resulting in AP-1 factor formation [42,121,124,127] c-myc oncogene C-myc, c-fos and c-myb oncogenes provide attractive targets for prevention of restenosis as they are involved in SMC proliferation and migration as well as deposition of extracellular matrix after post-vascular injury [128] Inflammatory diseases c-jun c-jun/AP-1 can be activated by many extracellular stimulators including viral infections and can initiate and enhance the transcription of antiviral agents, which can cause significant inflammation, and appears to have an important role in virus infections and replications. [129] iNOS iNOS is expressed only after induction by inflammatory mediators such as lipopolysaccharides, IL-1 and TNF-a [130,131] TGF-b1 TGF-b regulates cell proliferation, differentiation and immune reaction, one of the most important tasks being regulation of extracellular matrix accumulation; inhibition of mesangial TGF-b expression is one of therapeutic strategies to prevent renal fibrosis [132] GATA-3 Transcription factor GATA-3 plays an important role in Th2 cell activation, but also in the regulation of other cell types involved in bronchial asthma, including mast cells, eosinophils and epithelial cells [33,34,[133][134][135] CNS diseases XT-1 XT-1 is crucial for biosynthesis of glycosylated proteoglycans in the scar after spinal cord injury [136,137] HD gene Selective loss of neurons in HD is caused by abnormal expansion of the CAG triplet (> 36 repeats) of the HD gene techniques and chemotherapy.…”