高迁移率族蛋白B1修饰间充质干细胞对球囊损伤大鼠颈动脉的保护效应

Protective effects of mesenchymal stem cells modified by HMGB1 on balloon-induced carotid artery injury

目的探讨高迁移率族蛋白B1(high mobility group box1,HMGB1)修饰间充质干细胞(mesenchymal stem cells,MSC)对球囊损伤大鼠颈动脉的作用及分子机制。方法 HMGB1重组腺病毒(ad5GFP-HMGB1)转染MSC后,实时定量PCR、Western blot和免疫组化法检测MSC中HMGB1、血管内皮生长因子(VEGF)、增殖细胞核抗原(PCNA)的表达水平。构建球囊损伤大鼠颈动脉模型,Evans Blue染色评价损伤血管再内皮化;ELISA检测血清中肿瘤坏死因子-α(TNF-α)和C反应蛋白(CRP)水平。结果与对照组和空载ad5GFP组比较,HMGB1修饰增加MSC中VEGF和PCNA的mRNA和蛋白表达水平(P均<0.05)。Evans Blue染色显示,与对照组(11.35±2.25)和空载ad5GFP组(36.78±3.75)比较,ad5GFP-HMGB1组(47.66±9.11)损伤血管再内皮化面积明显增加,差异均具有统计学意义(P均<0.01)。ELISA结果显示,HMGB1修饰MSC能降低颈动脉损伤大鼠血清中TNF-α和CRP的表达水平(P均<0.05)。结论 HMGB1修饰MSC对大鼠颈动脉损伤具有保护效应,其机制可能与上调VEGF和PCNA表达及抑制炎症反应有关。HMGB1可能是MSC修复内皮过程中的一个新的治疗靶点。

Objective To explore the protective effects of mesenchymal stem cells （MSC）modified by highmobility group box 1 （HMGB1） on balloon-induced carotid artery injury.Methods MSC were infected by adenoviral serotype 5 encoding recombinant green fluorescent protein （GFP） gene and HMGB1 （ad5GFP-HMGB1）.The expression levels of HMGB 1,vascular endothelial growth factor （VEGF） and proliferating cell nuclear antigen （PCNA） were detected in HMGB1-modified MSC using real-time PCR,western blot and tissue immunohistochemical assays.A balloon-induced carotid artery injury model was established in rats.Reendothelialization was evaluated by Evans Blue staining,and the serum levels of tumor necrosis factor-α （TNF-α） and C-reactive protein （CRP） were assessed by ELISA assay.Results Compared with MSC group and ad5 GFP group,the expression levels of HMGB1,VEGF and PCNA were markedly increased in HMGBl-modified MSC （each P 〈 0.05）.The Evans Blue staining demonstrated that the reendothelialization areas （47.66 ± 9.11）of carotid artery injury were significantly increased,compared with MSC group （11.35 ± 2.25） and ad5 GFP group （36.78 ± 3.75） （each P 〈 0.01）.ELISA assay showed that the serum levels of TNF-α and CRP were decreased in group ad5GFP and ad5GFP-HMGB1 compared with MSC group（each P 〈 0.05）.Conclusions Transplantation of HMGB1-modified MSC exerts a protective effect on balloon-induced carotid artery injury,which may be associated with the up-regulation of VEGF and PCNA and inhibition of inflammatory response.HMGB1 may be a new therapeutic target in the process of MSC mediated endothelial repair.