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目的:阳离子磷酸胆碱聚合物(MPC30-DEA70)为非病毒类转基因载体,可与AMO-mi R-222络合,通过导管球囊系统将MPC30-DEA70/AMO-mi R-222基因复合物导入大鼠颈内动脉球囊损伤处,观察对血管平滑肌细胞增生及血管狭窄程度的影响。方法:90只雄性SD大鼠随机分为未损伤组、多聚赖氨酸(PLL组)、MPC30-DEA70/AMO-mi R-222组、PLL/AMO-mi R-222组、PLL/MPC30-DEA70组、AMO-mi R-222组、单纯损伤组、PLL载P/A=3:1组和P/A=5:1组,每组各10只。构建大鼠颈总动脉球囊损伤模型,予血管损伤段行PLL、MPC30-DEA70/AMO-mi R-222、PLL/AMO-mi R-222、PLL/MPC30-DEA70、AMO-mi R-222、PLL载P/A=3:1和P/A=5:1复合物的转运。4周后通过光学显微镜观察HE染色血管段组织形态学改变。Western blot法检测各组血管段p57Kip2、p27Kip1蛋白的表达情况。RT-PCR法检测各组血管段mi R222扩增情况。结果:光学显微镜下,多聚赖氨酸(PLL组)、裸MPC30-DEA70/AMO-mi R-222组、PLL/AMO-mi R-222组、PLL/MPC30-DEA70组、AMO-mi R-222组、MPC30-DEA70组可见内膜显著增生,新生内膜/中膜比值无组间差异,较PLL载P/A=3:1组和P/A=5:1组有组间差异,后两组比较无组间差异,未损伤组未见新生内膜增殖。Western blot法检测显示PLL载P/A=3:1组和P/A=5:1组P57kip2、P27kip1蛋白表达含量较未损伤组降低(P<0.05),较余六组增高(P<0.05),组间比较无显著差异(P>0.05)。RT-PCR法检测显示,mi R-222表达在未损伤组很低,PLL载P/A=3:1组和P/A=5:1组增高,余六组过表达(组间比较无显著差异)。结论:MPC30-DEA70可与AMO-mi R-222络合,有效抑制球囊损伤后血管mi R222表达,从而抑制新生内膜增生及血管狭窄。
Objective: The cationic phosphocholine polymer (MPC30-DEA70) is a non-viral genetically modified vector that can be complexed with AMO-mi R-222 and the MPC30-DEA70 / AMO-mi R-222 gene complex The rats were infused into the carotid artery of rabbits to observe the effect on vascular smooth muscle cell proliferation and vascular stenosis. Methods: Ninety male Sprague-Dawley rats were randomly divided into four groups: uninjured group, polylysine group (PLL group), MPC30-DEA70 / AMO-mi R-222 group, PLL / AMO-mi R- -DEA70 group, AMO-mi R-222 group, simple injury group, PLL contained P / A = 3: 1 group and P / A = 5: 1 group. The model of balloon injury in the common carotid artery of rats was established. The injured vessels were treated with PLL, MPC30-DEA70 / AMO-mi R-222, PLL / AMO-mi R- 222, PLL / MPC30-DEA70 and AMO-mi R-222 , PLL contained P / A = 3: 1 and P / A = 5: 1 transport of the complex. After 4 weeks, the morphological changes of HE staining vascular segments were observed by optical microscope. Western blot was used to detect the expression of p57Kip2 and p27Kip1 in each group. RT-PCR method was used to detect the amplification of mi R222 in each group. RESULTS: Under the light microscope, poly-L-lysine (PLL) group, naked MPC30-DEA70 / AMO-mi R-222 group, PLL / AMO-mi R-222 group, PLL / MPC30-DEA70 group, AMO-mi R -222 group, MPC30-DEA70 group showed significant intimal hyperplasia, neointimal / medial membrane ratio without group differences, compared with P / A A = 3: 1 group and P / A = 5: , No difference between the two groups after the comparison, no neointimal proliferation in the no injury group. Western blot showed that the expression of P57kip2 and P27kip1 in P / A = 3: 1 group and P / A = 5: 1 group were lower than those in non-injury group (P <0.05) ), No significant difference between the two groups (P> 0.05). The results of RT-PCR showed that the expression of mi R-222 was low in non-injured group, P / A = 3: 1 group and P / A = 5: 1 group in PLL group, and the other six groups were overexpressed Significant differences). CONCLUSION: MPC30-DEA70 can complex with AMO-mi R-222 and effectively inhibit the expression of mi R222 after balloon injury, thereby inhibiting neointimal hyperplasia and vascular stenosis.