Meng Zhaoyou, Gao Pan, Chen Lin, Peng Jing, Huang Jialu, Wu Min, Chen Kangning, Zhou Zhenhua
Department of Neurology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Department of Geratology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Mol Ther Nucleic Acids. 2017 Sep 15;8:123-131. doi: 10.1016/j.omtn.2017.06.010. Epub 2017 Jun 19.
The inhibition of inflammation and vascular smooth muscle cell (VSMC) proliferation is an ideal strategy to suppress intimal hyperplasia after percutaneous transluminal angioplasty (PTA). Evidence has indicated that overexpression of A20 suppresses neointima formation, but its low transfection efficiency limits its application. Hence, we upregulated A20 expression via transfection of rAd.ATF (recombinant adenovirus vector of artificial transcription factor) and rAd.A20 in rat carotid arteries after balloon dilatation (in vivo) and isolated VSMCs (in vitro). In vivo, we found that after rAd.ATF and rAd.A20 transfection, A20 expression was markedly increased, whereas proliferating cell nuclear antigen (PCNA) and nuclear factor κB p65 (NF-κBp65) protein levels were significantly decreased, and intimal hyperplasia and secretion of proinflammatory factors were significantly reduced when compared with empty vector and saline control groups. Most importantly, the rAd.ATF-treated group showed more significant inhibition on intimal hyperplasia and expression of PCNA than the rAd.A20-treated group. In vitro, compared with the control group, transfection of rAd.ATF and rAd.A20 significantly increased A20 expression, which upregulated the proliferator-activated receptor (PPAR) level for both mRNA and protein, and reduced migration and proliferation of VSMCs and lipopolysaccharide (LPS)-induced inflammation. Furthermore, the PPARα agonist GW6471 could partially restore the effect of A20 on VSMCs. Our findings indicate that the ATF of A20 inhibits neointimal hyperplasia and, therefore, constitutes a novel potential alternative to prevent restenosis.
抑制炎症和血管平滑肌细胞（VSMC）增殖是抑制经皮腔内血管成形术（PTA）后内膜增生的理想策略。有证据表明，A20的过表达可抑制新生内膜形成，但其低转染效率限制了其应用。因此，我们通过在大鼠颈动脉球囊扩张后（体内）和分离的VSMC（体外）中转染rAd.ATF（人工转录因子重组腺病毒载体）和rAd.A20来上调A20表达。在体内，我们发现转染rAd.ATF和rAd.A20后，A20表达明显增加，而增殖细胞核抗原（PCNA）和核因子κB p65（NF-κBp65）蛋白水平显著降低，与空载体和生理盐水对照组相比，内膜增生和促炎因子分泌显著减少。最重要的是，rAd.ATF治疗组对内膜增生和PCNA表达的抑制作用比rAd.A20治疗组更显著。在体外，与对照组相比，转染rAd.ATF和rAd.A20显著增加A20表达，上调增殖激活受体（PPAR）的mRNA和蛋白水平，并减少VSMC的迁移和增殖以及脂多糖（LPS）诱导的炎症。此外，PPARα激动剂GW6471可部分恢复A20对VSMC的作用。我们的研究结果表明，A20的ATF可抑制内膜增生，因此是预防再狭窄的一种新的潜在替代方法。
