转化生长因子-β1对斑块内新生血管生成及颈动脉斑块稳定性的影响及其机制

Effect of transforming growth factor-β1 on intraplaque angiogenesis and stability of carotid plaque and underlying mechanism

目的:观察靶向调控转化生长因子(TGF)-β1下游β-连环蛋白(β-catenin)/T细胞因子(TCF)与β-catenin/叉头框蛋白O1(FOXO1)信号的竞争抑制对颈动脉斑块内新生血管生成及斑块稳定性的影响。方法:收集CEA术后斑块组织( n=21),对斑块组织进行苏木精-伊红(HE)染色后据病理特征分出易损和稳定斑块,后对各组织中CD31、VE-Cadherin的表达部位及水平进行免疫组织化学(IHC)染色分析;ApoE -/-小鼠(T、TI、I、N 4组, n=12)制作颈动脉狭窄模型,设T组TGF-β1(每天50 μg/kg)+生理盐水(5 mg/kg);TI组TGF-β1(50 μg/kg)+ICG-001(5 mg/kg);I组生理盐水(50 μg/kg)+ICG-001(5 mg/kg);N组生理盐水(5.05 mg/kg)并每天相应处理连续2周,取颈动脉组织,对各组织行HE染色后据病理特征分出易损和稳定斑块,后用IHC染色及半定量评估各组织中CD31、VE-Cadherin的表达部位及水平。组间比较采用两独立样本 t检验或单因素方差分析。 结果:T组和TI组易损斑块率分别为60.00%、33.33%,均较N组的72.73%低;VE-Cadherin在T组与N组表达分别为0.156±0.007、0.191±0.005,差异有统计学意义( F=1.615, P<0.05),与TI组表达(0.133±0.010)比较差异有统计学意义( F=0.068, P<0.05);CD31在TI组和T组表达分别为0.119±0.004、0.136±0.010,差异有统计学意义( F=3.451, P<0.05)。IHC切片可见稳定斑块中CD31、VE-Cadherin标记含量低,且TI组含量低于其他各组。 结论:特异性阻断TGF-β下游的β-catenin/TCF可反向增强β-catenin/FoxO1对颈动脉粥样硬化斑块内新生血管的抑制作用,进而有利于稳定斑块。

Objective To investigate the effect of competitive inhibition ofβ-catenin/T-cell factor andβ-catenin/forkhead box protein O1(FoxO1)signal downstream of transforming growth factor(TGF)-β1 on angiogenesis and plaque stability in carotid plaque.Methods Plaque tissues of patients with carotid stenosis after CEA were collected(n=21).Vulnerable and stable plaques were identified by hematoxylin-eosin(HE)staining,and the expression sites and levels of CD31 and VE-Cadherin in each tissue were analyzed by immunohistochemistry(IHC).Model of carotid stenosis was established in ApoE-/-mice.Follwoing groups were set up(n=12/group).Group T was given TGF-β1(50μg/kg)+normal saline(5 mg/kg).Group TI was given TGF-β1(50μg/kg)+ICG-001(5 mg/kg).Group I was given normal saline(50μg/kg)+ICG-001(5 mg/kg).Group N was given normal saline(5.05 mg/kg).All groups were treated by their own treatment for 2 weeks,then the carotid tissues were taken.After HE staining,vulnerable and stable plaques were divided according to the pathological characteristics.The expression sites and levels of CD31 and VE-Cadherin in each tissue were detected by IHC.All data were analyzed by Imageplus 6.0 and SPSS 25.0 statistical software.Results The vulnerable rate in groups T and TI was 60.00%and 33.33%respectively,which was lower than in group N(72.73%).The expression of VE-Cadherin in groups T and N was 0.156±0.007 and 0.191±0.005 respectively,which was statistically significant(F=1.615,P<0.05).The expression of VE-Cadherin in group T was significantly higher than that in group TI(0.133±0.010,F=0.068,P<0.05).The expression of CD31 in groups TI and T was 0.119±0.004 and 0.136±0.010 respectively,which was statistically significant(F=3.451,P<0.05).The expression of CD31 and VE-Cadherin in stable plaque was low,and that in group TI was lower than in the rest groups.Conclusion Specific blockingβ-catenin/TCF downstream of TGF-βcan inversely enhance the inhibitory effect ofβ-catenin/FOXO1 on carotid atherosclerosis intraplaque angiogenesis,thereby having a positive impact on stabilizing plaque.