c-fos基因与静脉平滑肌表型转换的相关性研究

C-fos expression is correlated with phenotypic switching of vascular smooth muscle cells derived from venous varicosities

目的:探讨静脉曲张的组织学和细胞学改变以及c-fos上调与血管平滑肌细胞(VSMCs)表型转换的关系。方法:2019年1月至6月纳入复旦大学附属华东医院31例静脉曲张患者(曲张静脉组)和12例冠状动脉搭桥术的患者(正常静脉组)纳入研究,采集大隐静脉进行实验。实时定量聚合酶链反应、免疫组织化学方法检测c-fos的表达。原代培养VSMCs采用细胞计数试剂盒(CCK-8)法和划痕法检测VSMCs的增殖和迁移能力。采用SPSS 22.0软件进行 χ2检验或 t检验。 结果:曲张组静脉管腔扩张,管壁增厚,细胞排列紊乱。与对照组比较,曲张组c-fos和骨桥蛋白(OPN)的mRNA水平明显升高( t=4.872、6.221, P<0.05),且c-fos表达与OPN/α-平滑肌肌动蛋白(α-SMΑ)呈正相关( R2=0.509, P<0.01)。来自曲张组的VSMCs增殖(0.851±0.048比1.493±0.064, t=13.990, P<0.05)和迁移(0.403±0.032比0.708±0.033, t=9.335, P<0.05)能力显著增强。此外,曲张组VSMCs的c-fos蛋白表达显著上调( t=17.270, P<0.05),伴随着α-SMA的降低( t=3.329, P<0.05)和OPN的增加( t=5.990, P<0.05)。 结论:病变标本中c-fos的表达水平上调,同时表型标志物(OPN/α-SMA)也发生改变。曲张组原代培养的VSMCs具有增强的增殖和迁移能力。c-fos的上调可能在VSMCs表型转换中起作用,进而参与静脉曲张的发病机制。

Objective To investigate the histological and cytological changes in varicose veins(VVs).Methods Patients with VVs and patients undergoing coronary artery bypass were enrolled.Real-time quantitative polymerase chain reaction and immunohistochemistry were used to assess the mRNA and protein expression of c-fos,α-smooth muscle actin(α-SMA)and osteopontin(OPN).Primary VSMCs were isolated from both groups and cultured in vitro.SPSS 22.0 software was used to perform statistical analysis with theχ2 test or Student’s t-test.Results Significantly elevated mRNA levels of c-fos and osteopontin(OPN)were observed in the VV group(t=4.872,6.221,P<0.05),and a positive correlation was further demonstrated between the mRNA levels of c-fos and those of OPN/α-SMA(R2=0.509,P<0.01).The VSMCs derived from the VV group were more numerous and had a significantly higher migration speed(t=9.335,P<0.05).Moreover,the protein expression of c-fos was significantly upregulated in VSMCs derived from the VV group(t=17.270,P<0.05),and this change was accompanied by a decrease inα-SMA(t=3.329,P<0.05)and an increase in OPN(t=5.990,P<0.05).Conclusion C-fos upregulation might play a role in the phenotypic switching of VSMCs and subsequently participate in the pathogenesis of VVs.