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目的研究骨髓间充质干细胞(BMSCs)移植对染矽尘小鼠肺部早期炎症的影响。方法取5只无特定病原体级健康雄性C57BL/6小鼠,采用全骨髓贴壁法分离培养BMSCs。取30只同类小鼠随机分为对照组、二氧化硅组和BMSCs移植组3组,以气管暴露法建立动物模型,对照组小鼠予气管内一次性注射0.90%氯化钠溶液20.0μL;二氧化硅组和BMSCs移植组小鼠先予气管内一次性注射20.0μL质量浓度为250 g/L的二氧化硅混悬液,6 h后再分别经尾静脉输注500.0μL的0.90%氯化钠溶液或细胞密度为1×109/L的BMSCs悬液。造模后第7天处死小鼠,观察肺组织病理学改变情况,采用酶联免疫吸附实验检测小鼠血清中白细胞介素(IL)-1β、IL-6和IL-10水平,采用实时定量聚合酶链反应仪检测小鼠肺组织中上述细胞因子mRNA的表达情况。结果 BMSCs表面分子分化抗原(CD)29、CD34、CD90、CD105、CD106的阳性率分别为67.70%、0.12%、39.00%、37.10%和20.10%。肺组织病理学观察显示,二氧化硅组小鼠肺泡壁增厚,肺泡间隔增宽,肺泡结构破坏;BMSCs移植组肺组织结构无明显破坏,肺泡腔大小均匀,肺泡结构完整。二氧化硅组小鼠血清中IL-1β、IL-6水平以及肺组织中IL-1β、IL-6 mRNA相对表达水平分别高于对照组和BMSCs移植组(P<0.05),BMSCs移植组小鼠血清中IL-1β水平和肺组织中IL-1βmRNA相对表达水平均高于对照组(P<0.05)。各组小鼠血清中IL-10水平和肺组织中IL-10 mRNA相对表达水平分别比较,差异均无统计学意义(P>0.05)。结论 BMSCs可通过下调促炎因子IL-1β和IL-6的表达减轻染矽尘小鼠肺部早期炎性损伤。
Objective To investigate the effects of bone marrow mesenchymal stem cells (BMSCs) transplantation on the early lung inflammation in mice exposed to silica dust. Methods Five healthy male C57BL / 6 mice without specific pathogen were selected and BMSCs were isolated and cultured by whole bone marrow adherent method. Thirty mice of the same type were randomly divided into three groups: control group, silica group and BMSCs transplantation group. The animal model was established by tracheal exposure. In the control group, 20.0 μL of 0.90% sodium chloride solution was injected into the trachea. Silica group and BMSCs transplantation group were given intraperitoneal instillation of 20.0 μL of 250 g / L silica suspension. After 6 h, 500.0 μL of 0.90% chlorine Sodium solution or BMSCs suspension with a cell density of 1 × 10 9 / L. The mice were sacrificed on the 7th day after the model was established. The pathological changes of the lung tissue were observed. The levels of IL-1β, IL-6 and IL-10 in the serum of the mice were detected by enzyme-linked immunosorbent assay (ELISA) Polymerase chain reaction (PCR) was used to detect the mRNA expression of the above cytokines in lung tissue of mice. Results The positive rates of CD34, CD34, CD90, CD105 and CD106 on BMSCs were 67.70%, 0.12%, 39.00%, 37.10% and 20.10% respectively. Lung histopathology showed that alveolar wall thickening, alveolar septum broadening and alveolar structure destruction were found in the silica group. The lung tissue structure of the BMSCs transplantation group had no obvious damage, the size of the alveolar cavity was uniform and the alveolar structure was intact. The levels of IL-1β and IL-6 in sera of silica group and the relative expression of IL-1β and IL-6 mRNA in lung tissue were higher than those in control group and BMSCs transplantation group (P <0.05) The level of IL-1β in murine serum and IL-1β mRNA in lung tissue were higher than those in control group (P <0.05). There was no significant difference in serum IL-10 level and IL-10 mRNA relative expression level in each group (P> 0.05). Conclusion BMSCs can reduce the early inflammatory injury in the lungs of mice exposed to silica dust by down-regulating the expression of proinflammatory cytokines IL-1β and IL-6.