摘要
目的 探讨核因子κB/生存素信号通路在大鼠颈动脉球囊损伤模型内膜增生中的作用.方法 构建核因子κB siRNA慢病毒载体（滴度1×108 TU/ml）,并在33只SD大鼠中建立颈动脉球囊损伤模型.建模后将SD大鼠分为阴性对照组（n=11）、核因子κB siRNA转染组（n=11）和核因子κB siRNA转染＋YM155（生存素抑制剂）组（n=11）,同时以损伤对侧的正常颈动脉作为正常对照组（n=11）.7d后各组分别处死5只SD大鼠并取材,以RT-PCR检测核因子κB及生存素mRNA水平.28 d后各组分别处死6只SD大鼠并取材,苏木精-伊红染色后比较内膜增生程度,以免疫组织化学法检测内膜增殖细胞核抗原（PCNA）及中膜α-SM-actin的表达.结果 （1）术后7d,阴性对照组核因子κB及生存素mRNA水平均高于正常对照组（P均＜0.05）;核因子κB siRNA转染组核因子κB mRNA水平低于阴性对照组（P＜0.05）,而与核因子κB siRNA转染＋YM155组比较差异无统计学意义（P＞0.05）;核因子κB siRNA转染组生存素mRNA水平低于阴性对照组（P＜0.05）,而高于核因子κB siRNA转染＋YM155组（P＜0.05）.（2）术后28 d,与正常对照组比较其余3组均有不同程度的内膜增生.阴性对照组、核因子κB siRNA转染组、核因子κB siRNA转染＋YM155组中膜面积差异无统计学意义（P＞0.05）.阴性对照组、核因子κB siRNA转染组、核因子κB siRNA转染＋YM155组的内膜面积分别为（0.13 ±0.01）、（0.11 ±0.01）和（0.09 ±0.01）mm2,各组间差异均有统计学意义（P均＜0.05）;内膜面积/中膜面积分别为1.55±0.07、0.92 ±0.08和0.76±0.06,各组间差异均有统计学意义（P均＜0.05）;剩余狭窄率分别为（58.71±0.02）、（32.13±0.05）和（26.42±0.03）％,各组间差异均有统计学意义（P均＜0.05）;内膜PCNA阳性表达率分别为（45.32±7.21）、（36.54±6.42）和（28.57±6.31）％,各组间差异均有统计学意义（P均＜0.05）;中膜α-SM-action平均光密度值分别为0.055±0.006、0.072±0.011和0.084±0.008,各组间差异均有统计学意义（P均＜0.05）.结论 抑制核因子κB基因表达可抑制损伤内膜的增生,其作用机制可能与抑制生存素而减轻血管平滑肌细胞增殖有关.
Objective To investigate the role of NF-κB/survivin signal pathway in the intima hyperplasia of rat carotid balloon injury restenosis model.Methods NF-κB siRNA lentivirus vector（titer was 1 × 108 TU/ml）was established.Carotid balloon injury restenosis model was made in 33 SD rats.The rats were divided into 4 groups according to different processing methods,including negative control （NC） group （n =11）,NF-κB siRNA group（n =11）,NF-κB siRNA ＋ YM155 （survivin inhibitor） （n =11）,the uninjured carotid artery served as the normal control group（n =1 1）.After 7 days,the carotid sample（n =5 each group）were harvested to detect the NF-κB and survivin mRNA expression by RT-PCR.The carotid sample were harvested on 28 days （n =6 each group） for HE staining and measuring intima hyperplasia.Immunohistochemical method was also used to detect the expression of intima proliferation cell nuclear antigen （PCNA） and media α-SM-actin.Results （1） After 7 days,NF-κB and survivin mRNA expression was significant higher in NC group than in normal control group （P ＜ 0.05）,the NF-κB mRNA expression was significantly lower in NF-κB siRNA group than in NC group （P ＜ 0.05） and similar between NF-κB siRNA group and NF-κB siRNA ＋ YM155 group.The survivin mRNA expression was significantly lower in NF-κB siRNA group compared to NC group （P ＜ 0.05） and significantly higher in NF-κB siRNA group than in NF-κB siRNA ＋ YM155 group （P ＜0.05）.（2） After 28 days,intima hyperplasia was observed in NC （0.13 ±0.01）,NF-κB siRNA （0.11 ±0.01） and NF-κB siRNA ＋YM155 group（0.09 ±0.01）mm2（P ＜ 0.05）.Media area was similar among NC group,NF-κB siRNA group and NF-κB siRNA ＋ YM155 group （P ＞ 0.05）.L/M ratio was gradually reduced among NC group（1.55 ± 0.07）,NF-κB siRNA group（0.92 ± 0.08）,NF-κB siRNA ＋ YM155 group（0.76 ±0.06,all P ＜0.05）.Similar results were found in the residual restenosis rate：NC group （58.71 ± 0.02） ％,NF-κB siRNA group （32.13 ± 0.05） ％,NF-κB siRNA ＋ YM155 group （26.42 ± 0.03） ％ （all P ＜ 0.05） and expression of vascular smooth muscle cell PCNA：NC group（45.32 ± 7.21） ％,NF-κB siRNA group（36.54 ± 6.42） ％,NF-κB siRNA ＋ YM155 group（28.57 ± 6.31）％ （all P ＜ 0.05）.On the contrary,the IOD of α-SM-actin in media increased gradually：NC group （0.055 ± 0.006）,NF-κB siRNA group （0.072 ± 0.011）,NF-κB siRNA ＋ YM155 group（0.084 ±0.008,all P ＜ 0.05）.Conclusion Inhibiting NF-κB expression can significant decrease intima hyperplasia in this model,and this effect may be mediated by inhibiting survivin and reducing the proliferation of vascular smooth muscle cells.
出处
《中华心血管病杂志》
CAS
CSCD
北大核心
2015年第3期248-253,共6页
Chinese Journal of Cardiology
基金
湖北省自然科学基金（2010CDB03601）
