摘要
目的阳离子磷酸胆碱聚合物(MPC30-DEA70)负载反义miR-146a寡核苷酸(AMO-miR-146a)转染大鼠颈总动脉球囊损伤处血管平滑肌细胞(VSMC),观察其对血管内膜增生的影响。方法 60只SD大鼠完全随机化分为未损伤组、单纯损伤组、多聚赖氨酸(PLL)组、AMO-miR-146a组、PLL载MPC30-DEA70/AMO-miR-146a复合物(P/M=3:1组和P/M=5:1组),每组10只。通过光学显微镜观察4周后大鼠颈总动脉组织形态学变化,q-PCR法检测miR-146a的表达以及应用Western blot法检测增殖细胞核抗原蛋白(PCNA)、NFκBp65的表达情况。结果苏木精-伊红染色,光镜下显示,除未损伤组外,单纯损伤组、PLL组、AMO-miR-146a组、P/M=3:1组、P/M=5:1组大鼠颈总动脉血管新生内膜有不同程度的增生,而中膜面积无明显改变;与单纯损伤组比较,PLL组、AMO-miR-146a组新生内膜面积差异无统计学意义(P>0.05),P/M=3:1组和P/M=5:1组新生内膜面积明显减少,差异有统计学意义(P<0.05),后两组组间比较差异无统计学意义(P>0.05);q-PCR及Western blot检测显示P/M=3:1组和P/M=5:1组miR-146a及目的蛋白PCNA、NFκBp56的表达较未损伤组高,差异有统计学意义(P<0.05),但明显低于单纯损伤组、PLL组、AMO-miR-146a组,差异有统计学意义(P<0.05),而P/M=3:1组和P/M=5:1组组间miR-146a、PCNA、NFκBp56的表达差异无统计学意义(P>0.05);单纯损伤组、PLL组、AMO-miR-146a组组间miR-146a、PCNA、NFκBp56的表达差异无统计学意义(P>0.05)。结论 MPC30-DEA70可与AMO-miR-146a络合,借助PLL进入VSMC,有效抑制球囊损伤后VSMC的miRNA-146a的表达,使PCNA、NFκBp56下调,抑制新生内膜增生,防止血管狭窄。
Objective To investigate the influence of the delivery of phosphorylcholine copolymer MPC30-DEA70 carrying antisense mediated oligonucleotide miR-146 a gene into vascular smooth muscle cells（VSMC） on the degree of neointimal hyperplasia after balloon injury in rats. Methods A total of 60 rats were randomly divided into six groups as follows： no-injury group, pure balloon injury group, polylysine（PLL） group, AMO-miR-146 a group, PLL-carrier P/M = 3：1 group and P/M= 5：1 group, with 10 rats in each group. The morphological changes of common carotid arteries in rats after 4 weeks were observed by optical microscope. q-PCR method was used to detect the expression of miR-146 a, and Western blot was adopted to detect the expression of proliferating cell nuclear antigen protein（PCNA） and NFκBp65. Results Except for the no-injury group, the other experimental groups showed varying degrees of neointimal hyperplasia and vascular stenosis with HE staining, but the medial area had no significant change. Compared with the pure balloon injury group, the polylysine group and AMO-miR-146 a group had no significant difference in new neointima area（P〈0.05）; P/M=3：1 group and P/M=5：1 group were reduced significantly（P〈0.05）, but there was no significant difference between P/M=3：1 group and P/M=5：1 group（P〉0.05）. The detection of q-PCR and Western blot methods showed the expressions of miR-146 a and the protein expression contents of PCNA and NFκBp56 in the PLL-carrier P/M=3：1 and P/M=5：1groups were significant higher than those in the no-injury groups（P〈0.05）, but significantly lower than those in the pure balloon injury group, polylysine（PLL） group, and AMO-miR-146 a group（P〈0.05）. There was no significant difference between P/M=3：1 group and P/M=5：1 group in the expression of miR-146 a, PCNA, NFκBp56（P〈0.05）. There was also no significant difference among pure balloon injury group, polylysine group and AMO-miR-146 a group on the expressions of miR-146 a,
出处
《海南医学》
CAS
2017年第11期1724-1727,共4页
Hainan Medical Journal
