Department of Cardiothoracic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
Department of Cardiothoracic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, 200233, China.
Int J Biol Sci. 2017 Oct 17;13(10):1309-1319. doi: 10.7150/ijbs.20254. eCollection 2017.
Vein graft failure due to neointimal hyperplasia remains an important and unresolved complication of cardiovascular surgery. microRNA-21 (miR-21) plays a major role in regulating vascular smooth muscle cell (VSMC) proliferation and phenotype transformation. Thus, the purpose of this study was to determine whether adenovirus-mediated miR-21 sponge gene therapy was able to inhibit neointimal hyperplasia in rat vein grafts. Adenovirus-mediated miR-21 sponge was used to inhibit VSMC proliferation and neointimal formation . To improve efficiency of delivery gene transfer to the vein grafts, 20% poloxamer F-127 gel was used to increase virus contact time and 0.25% trypsin to increase virus penetration. Morphometric analyses and cellular proliferation were assessed for neointimal hyperplasia and VSMC proliferation. miR-21 sponge can significantly decrease the expression of miR-21 and proliferation in cultured VSMCs. Cellular proliferation rates were significantly reduced in miR-21 sponge-treated grafts compared with controls at 28 days after bypass surgery (14.6±9.4 vs 34.9±10.8%, =0.0032). miR-21 sponge gene transfer therapy reduced the intimal/media area ratio in vein grafts compared with the controls (1.38±0.08 vs. 0.6±0.10, <0.0001). miR-21 sponge treatment also improved vein graft hemodynamics. We further identified that phosphatase and tensin homolog (PTEN) is a potential target gene that was involved in the miR-21-mediated effect on neointimal hyperplasia in vein grafts. Adenovirus-mediated miR-21 sponge gene therapy effectively reduced neointimal formation in vein grafts. These results suggest that there is potential for miR-21 sponge to be used to prevent vein graft failure.
静脉移植物失功导致的内膜增生仍然是心血管手术的一个重要且尚未解决的并发症。微小 RNA-21（miR-21）在调节血管平滑肌细胞（VSMC）增殖和表型转化方面发挥着重要作用。因此，本研究旨在确定腺病毒介导的 miR-21 海绵基因治疗是否能够抑制大鼠静脉移植物的内膜增生。使用腺病毒介导的 miR-21 海绵抑制 VSMC 增殖和内膜形成。为了提高基因转移到静脉移植物的效率，使用 20%泊洛沙姆 F-127 凝胶增加病毒接触时间，使用 0.25%胰蛋白酶增加病毒穿透。对新生内膜增生和 VSMC 增殖进行形态计量学分析和细胞增殖评估。miR-21 海绵可显著降低培养的 VSMC 中 miR-21 的表达和增殖。与对照组相比，旁路手术后 28 天 miR-21 海绵处理的移植物中细胞增殖率显著降低（14.6±9.4 对 34.9±10.8%，=0.0032）。与对照组相比，miR-21 海绵基因转染治疗降低了静脉移植物的内膜/中膜面积比（1.38±0.08 对 0.6±0.10，<0.0001）。miR-21 海绵治疗还改善了静脉移植物的血流动力学。我们进一步鉴定出磷酸酶和张力蛋白同源物（PTEN）是 miR-21 介导的静脉移植物内膜增生的潜在靶基因。腺病毒介导的 miR-21 海绵基因治疗可有效减少静脉移植物的新生内膜形成。这些结果表明，miR-21 海绵具有预防静脉移植物失功的潜力。
