浅表血栓性静脉炎管壁平滑肌细胞表型与功能变化

Phenotypic and functional transformation in smooth muscle cells derived from a superficial thrombophlebitis-affected vein wall

目的:探讨浅表性血栓性静脉炎(ST)、曲张大隐静脉(VGSVs)和正常大隐静脉(NGSVs)管壁平滑肌细胞(SMCs)之间是否存在差异性。方法:收集2019年1月至2020年5月中国人民解放军陆军第80集团医院ST和VGSVs及NGSVs标本,分离和培养SMCs。检测3组静脉B细胞淋巴瘤/白血病-2相关X蛋白(bax)、B细胞淋巴瘤/白血病-2(bcl-2)、半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3、基质金属蛋白酶(MMP)-2、MMP-9、组织基质金属蛋白酶抑制剂(TIMP)-1、TIMP-2表达。多组间比较采用单因素方差分析事后多重检验法。结果:血栓血管平滑肌细胞(TVSMCs)组bax、Caspase-3 mRNA(0.375±0.049、0.411±0.074)和蛋白表达(0.866±0.150、0.154±0.049)低于曲张静脉血管平滑肌细胞(VVSMCs)组(0.611±0.084、0.757±0.083、1.617±0.182、0.488±0.077)和正常血管平滑肌细胞(NVSMCs)组(2.441±0.137、0.925±0.081),差异有统计学意义(F=468.690、311.856、355.305、417.576,P<0.05);TVSMCs组bcl-2 mRNA[(2.228±0.108)倍]和蛋白表达(2.072±0.160)高于VVSMCs组(1.514±0.177、1.334±0.165)和NVSMCs组(0.385±0.085),差异有统计学意义(F=387.963、518.918,P<0.05);TVSMCs组MMP-2、MMP-9、TIMP-1、TIMP-2 mRNA[(2.634±0.172)、(1.994±0.161)、(2.493±0.178)、(2.362±0.143)倍]和蛋白表达(1.543±0.113、0.583±0.081、0.682±0.064、0.859±0.053)高于VVSMCs组[(1.386±0.127)、(1.373±0.145)、(1.920±0.123)、(1.869±0.140)倍、1.124±0.107、0.316±0.081、0.460±0.057、0.627±0.065]和NVSMCs组(0.468±0.074、0.125±0.047、0.154±0.046、0.254±0.045),差异有统计学意义(F=684.718、235.159、541.240、530.767、426.631、149.234、318.636、443.196,P<0.05)。结论:TVSMCs呈明显的去分化现象,提示TVSMCs在疾病进程中,其表型和功能存在的差异性,推测可能与血栓所致的静脉高压和炎症等因素有关。

Objective:This study tested the hypothesis whether relationships exist among smooth muscle cells(SMCs)derived from superficial thrombophlebitis(ST),varicose great saphenous veins(VGSVs),and normal great saphenous veins(NGSVs).Methods:ST,VGSVs,and NGSVs were collected from patients in the 80th Group Army Hospital of the People’s Liberation Army from January 2019 to May 2020.SMCs were isolated and cultured.B cell lymphoma/leukemia-2 associated X protein(bax),B cell lymphoma/leukemia-2(bcl-2),cysteinyl aspartate-specific protease(Caspase)-3,matrix metalloproteinases(MMP)-2,MMP-9,tissue inhibitor of metalloproteinases(TIMP)-1,and TIMP-2 mRNA and protein expressions were detected.Statistical analysis of each variable among the three or more different samples was carried out using one-way ANOVA,with the Student-Newman-Keuls method.Results:Bax and Caspase-3 mRNA and protein expression were decreased,while bcl-2 mRNA and protein expression were increased in the thrombophlebitic vascular smooth muscle cells(TVSMCs,0.375±0.049,0.411±0.074,0.866±0.150,0.154±0.049)vs.the varicose vascular smooth muscle cells(VVSMCs,0.611±0.084,0.757±0.083,1.617±0.182,0.488±0.077)and the normal vascular smooth muscle cells(NVSMCs,2.441±0.137;0.925±0.081,F=468.690、311.856、355.305、417.576、387.963、518.918,P<0.05).MMP-2,MMP-9,TIMP-1,and TIMP-2 mRNA and protein expression were significantly increased in the TVSMCs(2.634±0.172,1.994±0.161,2.493±0.178,2.362±0.143,1.543±0.113,0.583±0.081,0.682±0.064,0.859±0.053)vs.the VVGSVs(1.386±0.127,1.373±0.145,1.920±0.123,1.869±0.140,1.124±0.107,0.316±0.081,0.460±0.057,0.627±0.065)and the NVSMCs(0.468±0.074,0.125±0.047,0.154±0.046,0.254±0.045,F=684.718、235.159、541.240、530.767、426.631、149.234、318.636、443.196,P<0.05).Conclusion:SMCs derived from ST are more dedifferentiated.These results suggest that the phenotypic and functional differences could be related to the presence of high venous pressure and inflammation due to thrombosis during the disease course among SMCs derived from ST,VGSVs,and NGSVs.