脐带间充质干细胞在肺移植大鼠原发性移植肺失功中的治疗作用及机制研究

Therapeutic effect and mechanism of umbilical cord mesenchymal stem cells in rats with primary graft dysfunction after lung transplantation

  • 摘要:
    目的  探讨脐带间充质干细胞(UC-MSC)在大鼠原发性移植肺失功中的治疗作用及机制。
    方法  24只Lewis雄性大鼠随机分为供体和受体,各12只。受体分为3组:空白对照组、阴性对照组和治疗组,每组4只。肺移植术后72 h取材观察移植肺的颜色、大小、质地,通过micro-CT评估各组大鼠移植肺的通气情况及实变进展。收集受体大鼠血浆、移植肺组织和肺泡灌洗液样本,测定肺组织湿/干比以了解肺水肿程度,苏木素-伊红(HE)染色评估肺组织损伤程度,脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记(TUNEL)染色评估细胞凋亡情况,检测肺组织髓过氧化物酶(MPO)活性,酶联免疫吸附试验(ELISA)检测血浆及肺泡灌洗液中的白细胞介素(IL)-6、IL-10和肿瘤坏死因子(TNF)-α水平。
    结果  阴性对照组移植肺外观颜色与自体肺差异较大,治疗组与空白对照组移植肺与自体肺外观颜色几乎一致。与阴性对照组相比,治疗组肺泡内渗出减少,气道上皮细胞结构更为完整。空白对照组未见肺泡内渗出,气道及肺泡结构保持正常。治疗组气道上皮细胞凋亡率、肺组织湿/干比和MPO活性均低于阴性对照组(均为P<0.05)。治疗组肺泡灌洗液IL-6和TNF-α水平低于阴性对照组,IL-10水平高于阴性对照组和空白对照组(均为P<0.05)。各组间血浆中细胞因子水平差异均无统计学意义(均为P>0.05)。
    结论  UC-MSC能够通过减少肺组织细胞凋亡率和抑制炎症反应,有效缓解大鼠原发性移植肺失功的严重程度。

     

    Abstract:
    Objective To explore the therapeutic effect and mechanism of umbilical cord mesenchymal stem cells (UC-MSC) in rats with primary graft dysfunction after lung transplantation.
    Methods Twenty-four male Lewis rats were randomly divided into donor and recipient groups, with 12 rats in each group. The recipients were further divided into 3 groups: blank control group, negative control group, and treatment group, with 4 rats in each group. The color, size and texture of the transplanted lungs were observed 72 h after lung transplantation. The ventilation status and progression of consolidation in the transplant lungs of rats in each group were evaluated by micro-CT. Plasma, transplant lung tissue and alveolar lavage fluid samples of recipient rats were collected. The wet/dry ratio of lung tissue was measured to evaluate the degree of pulmonary edema. Hematoxylin-eosin (HE) staining was used to evaluate the degree of lung tissue damage. Terminal deoxyribonucleic acid transferase mediated dUTP nick end labeling (TUNEL) staining was used to evaluate cell apoptosis. Myeloperoxidase (MPO) activity in lung tissue was detected, and enzyme-linked immunosorbent assay (ELISA) was used to detect interleukin (IL)-6, IL-10 and tumor necrosis factor (TNF)-α levels in plasma and alveolar lavage fluid.
    Results The appearance of the transplant lungs in the negative control group was significantly different from that of the autologous lungs, while the transplant lungs in the treatment group were almost identical in color to the autologous lungs compared to the blank control group. Compared with the negative control group, the treatment group showed reduced alveolar exudate and more intact airway epithelial cell structure. No alveolar exudate was observed in the blank control group, and the structure of the airways and alveoli remained normal. The treatment group had lower apoptosis rate of airway epithelial cells, lung tissue wet/dry ratio, and MPO activity compared to the negative control group (all P < 0.05). The levels of IL-6 and TNF-α in the bronchoalveolar lavage fluid of the treatment group were lower than those in the negative control group, while the level of IL-10 was higher than that in the negative control group and the blank control group (all P < 0.05). There were no statistically significant differences in the levels of cytokines in plasma among each group (all P > 0.05).
    Conclusions UC-MSC may effectively alleviate the severity of primary graft dysfunction in rats by reducing the apoptosis rate of cells in lung tissue and inhibiting inflammatory responses.

     

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