骨髓间充质干细胞移植后颈动脉损伤球囊核转录因子κBp65和增殖细胞核抗原的表达

背景：炎症反应在经皮腔内冠状动脉支架置入后再狭窄的发生中起了重要作用。近年的研究发现,骨髓间充质干细胞具有较强的免疫调节特性。然而骨髓间充质干细胞对损伤血管炎症反应影响的报道较少。 目的：观察骨髓间充质干细胞移植对兔颈动脉球囊损伤后核转录因子κBp65和增殖细胞核抗原表达的影响,分析其与内膜增殖作用的相关性。 方法：制作颈动脉粥样硬化狭窄兔模型36只,随机分成2组,骨髓间充质干细胞移植组球囊损伤后接受4’,6-二脒基-2-苯基吲哚标记的骨髓间充质干细胞移植,对照组球囊损伤后接受等量的PBS液注射。造模前和细胞移植后7,14,28 d分别抽取外周血,ELIAS法检测血清肿瘤坏死因子α和白细胞介素6的质量浓度;细胞移植后14 d取血管组织,采用免疫组织化学分析法测定局部组织核转录因子κBp65和增殖细胞核抗原的表达,细胞移植后28 d对血管组织行苏木精-伊红染色检测血管形态学变化。 结果与结论：细胞移植后14 d 骨髓间充质干细胞移植组损伤血管局部核转录因子κBp65和增殖细胞核抗原的表达均较对照组明显减少(P < 0.05)。细胞移植后7,14,28 d骨髓间充质干细胞移植组血清肿瘤坏死因子α和白细胞介素6质量浓度均较对照组显著降低(P < 0.05)。细胞移植后28 d 骨髓间充质干细胞移植组新生内膜面积、新生内膜/中膜面积及血管狭窄率均较对照组显著降低(P < 0.05)。提示骨髓间充质干细胞可能通过抑制损伤血管核转录因子κBp65活性,减轻炎症反应进而下调增殖细胞核抗原的表达,对抑制损伤血管新生内膜的增殖可能发挥有益作用。

Expressions of nuclear factor κBp65 and proliferating cell nuclear antigen in injured carotid artery balloon after bone marrow mesenchymal stem cells transplantation

BACKGROUND:Inflammatory reaction plays an important role in the incidence of restenosis after percutaneous transluminal coronary intervention. Recent studies found that bone marrow mesenchymal stem cells (BMSCs) have strongly characteristics of immunological regulation. However, BMSCs on the inflammatory reaction in vascular injury is rarely reported. OBJECTIVE:To investigate effects of BMSCs transplantation on expression of nuclear factor κBp65 (NF-κBp65) and proliferating cell nuclear antigen (PCNA) after carotid artery balloon injury of rabbits, and to analyze its correlation with intimal proliferation. METHODS:Thirty-six rabbits were prepared for carotid artery atherosclerotic stenosis models and were randomly divided into the control group and BMSCs transplantation group. BMSCs pre-labled by DAPI was infused into rabbits by the ear vein in the BMSCs transplantation group, and the same amount of PBS solution was infused in the control group. The plasma tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels were detected by ELIAS in prior to model preparation and at 7, 14, 28 days after transplantation. The NF-κBp65 and PCNA expressions in vessels tissues were detected by immunohistochemical analysis at 14 days after transplantation, and vascular morphological changes were determined by hematoxylin-eosin staining at 28 days after transplantation. RESULTS AND CONCLUSION: Theexpression of NF-κBp65 and PCNA in the BMSCs transplantation group decreased significantly compared with control group at 14 days after transplantation (P < 0.05). At 7, 14, and 28 days after transplantation, the plasma TNF-α and IL-6 levels in the BMSCs transplantation group were significantly lower than those in the control group (P  < 0.05). The intimal area, the ratio of the intima/media area and the luminal stenosis ratio were significantly decreased in the BMSCs transplantation group than control group at 28 days after transplantation (P  < 0.05). BMSCs is capable to decrease the inflammatory reaction of injured vessels and down-regulate PCNA expression by inhibiting NF-κBp65 in injured vessels, and BMSCs may be good for inhibition of injured vessels intimal proliferation.