| 程 龙,李 志,王晓黎,冯 辉,燕 军,张卓琦,杨 煜,王志荣,徐 晤.MPC30-DEA70载AMO-miR-222对大鼠颈总动脉球囊损伤后血管狭窄的影响[J].,2017,17(1):36-41 |
| MPC30-DEA70载AMO-miR-222对大鼠颈总动脉球囊损伤后血管狭窄的影响 |
| Effects of AMO-miR-222 Carried by MPC30-DEA70 on Rats' Hemadostenosis after Common Arteria Carotis Balloon Injury |
| 投稿时间：2016-04-30 修订日期：2016-05-26 |
| DOI：10.13241/j.cnki.pmb.2017.01.009 |
| 中文关键词: 磷酸胆碱聚合物 反义miR-222 内膜增生 血管狭窄 |
| 英文关键词: Phosphorylcholine Polymer Antisense Mir-222 Intimal Hyperplasia Hemadostenosis |
| 基金项目:国家自然科学基金项目（30800219，30670557）；江苏省高校自然科学基金项目（08KJD320013）；徐州医学院院长专项人才基金项目（07KJZ26） |
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| 中文摘要: |
| 摘要 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-miR-222络合，通过导管球囊系统将MPC30-DEA70/AMO-miR-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-miR-222组、PLL/AMO-miR-222组、PLL/MPC30-DEA70组、AMO-miR-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-miR-222、PLL/AMO-miR-222、PLL/MPC30-DEA70、AMO-miR-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miR222扩增情况。结果：光学显微镜下，多聚赖氨酸（PLL组）、裸MPC30-DEA70/AMO-miR-222组、PLL/AMO-miR-222组、PLL/MPC30-DEA70组、AMO-miR-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，miR-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-miR-222络合，有效抑制球囊损伤后血管miR222表达，从而抑制新生内膜增生及血管狭窄。 |
| 英文摘要: |
| ABSTRACT Objective: As a non-viral transgenic carrier, the cationic phosphorylcholine polymer (MPC30-DEA70) can form a complexation with AMO-miR-222. The MPC30-DEA70/AMO-miR-222 gene compound is imported through catheter-balloon system in- to places where rats' internal arteria carotid balloon injury occurs to observe their effects on vascular smooth muscle cell proliferation and hemadostenosis. Methods: 90 male SD rats are randomly divided into 9 groups with 10 rats per group, as the groups respectively are: the uninjured group, polylysine group (PLL group), nude MPC30-DEA70/AMO-miR-222 group, PLL/AMO-miR-222 group, PLL/MPC30- DEA70 group, AMO-miR-222 group, only injured group, PLL-loaded P/A =3:1 group and P/A=5:1 group. Rats' common arteria carotis balloon injury model is built to transfer PLL, MPC30-DEA70/AMO-miR-222, PLL/AMO-miR-222, PLL/MPC30-DEA70, AMO-miR- 222, MPC30-DEA70, PLL-loaded P/A=3:1 and P/A=5:1 compounds for injured vascular. Four weeks later, histologic changes of HE stained blood vessel will be observed by optical microscope. The proteins expressions of p57Kip2 and p27Kip1 in blood vessels of those groups are detected by Western blot method. Amplification of miR222 in blood vessels of those groups is detected by RT-PCR method. Results: Under optical microscope, remarkable intimal hyperplasia can be observed in the polylysine (PLL) group, nude MPC30- DEA70/AMO-miR-222 group, PLL/AMO-miR-222 group, PLL/MPC30-DEA70 group, AMO-miR-222 and MPC30-DEA70, while the ratio of neointima/medial membrance among those groups shows no inter-group difference; however, compared with that of PLL-loaded P/A=3:1 and P/A=5:1 two groups, which are identical between the two groups, the ratio shows no difference; no neointimal hyperplasia is observed in the uninjured group. According to the Western blot method, compared with the uninjured group (P<0.05), the expressed con- tents of protein P57kip2 and P27kip1 in PLL loaded P/A=3:1 group and P/A=5:1 group are lower but higher than that of the rest six groups (P<0.05), while there is no remarkable difference between these two groups. As shown by the detection results of RT-PCR method, the expressed contents of miR-222 in the injured group is low; it is higher in PLL-loaded P/A=3:1 group and P/A=5:1 group, while too much of it is expressed in the rest six groups (there is no remarkable intergroup difference). Conclusion: MPC30-DEA70 and AMO-miR-222 can form a complexation together and effectively inhibit the expression of miR222 in blood vessel after balloon injury, so that inhibit neointimal hyperplasia and hemadostenosis can be inhibited. |
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