【摘要】 背景:近年来,间充质干细胞在器官或组织修复中展现了良好的前景,因此,基于间充质干细胞可塑性的细胞移植为体外循环后肺损伤的细胞再生及功能恢复提供了可能。目的:探讨外源性移植人羊膜问充质干细胞对犬体外循环缺血再灌注肺损伤的影响,从炎性因子角度初步探讨其作用机制。方法:健康成年杂种犬18只,随机分为空白组(建立体外循环模型,主动脉不阻断+股静脉注射1 mL生理盐水)、对照组(体外循环+主动脉阻断1h+主动脉开放15 min后股静脉注射1 mL生理盐水)、实验组(体外循环+主动脉阻断1 h+主动脉开放15 min后股静脉注射含2×10~7人羊膜间充质干细胞的生理盐水1 mL),每组6只。分别在体外循环前(T1)、主动脉开放后15 min(T2)、开放后1h(T3)、开放后2h(T4)、开放后3h(T5)5个时间点,各取动脉血2 mL计算氧合指数和呼吸指数,取静脉血8 mL分别用ELISA法检测血清肿瘤坏死因子α、基质金属蛋白酶9、白细胞介素8、白细胞介素10水平,Western Blot印迹法检测肺组织NF-κB蛋白表达,同时光镜下观察各组肺组织病理学变化。结果与结论:①与对照组比较,实验组氧合指数在人羊膜问充质干细胞移植后2,3 h显著升高,实验组呼吸指数在人羊膜问充质干细胞移植后1,2,3 h显著下降。②与对照组同时间点相比,实验组肿瘤坏死因子α、白细胞介素8以及基质金属蛋白酶9呈明显下降趋势,白细胞介素10有所回升。③与空白组相比,对照组开放后3 h肺组织NF-κB蛋白明显增强,静脉注入人羊膜间充质干细胞后肺组织NF-κB表达较对照组显著降低。④实验组在主动脉开放后,病理改变明显轻于对照组,可见少量间隙内渗出红细胞与中性粒细胞,微血管扩张、充血。⑤结果表明,人羊膜问充质干细胞静脉移植可通过下调促炎因子肿瘤坏死因子α、白细胞介素8,降低基质金属蛋白酶9水平,上调抗炎因子白细胞介素10水平,抑制肺组织NF-κB活化,减轻体外循环缺血再灌注相关肺损伤。更多还原

【Abstract】 BACKGROUND: In recent years, mesenchymal stem cells exhibit a good prospect in organ or tissue repair and therefore, and therefore, cell transplantation based on mesenchymal stem cell plasticity can promote cell regeneration and functional recovery from lung injury after cardiopulmonary bypass. OBJECTIVE: To investigate the effects of human amnion-derived mesenchymal stem cells(h AMSCs) transplantation on ischemia-reperfusion-induced acute lung injury in dogs after cardiopulmonary bypassand its mechanism for regulating inflammatory cytokines. METHODS: Eighteen adult healthy mongrel dogs were randomly divided into three groups(n=6 per group): black group(cardiopulmonary bypass with 1 m L physiological saline injection via the femoral vein without blocking the aorta), control group(cardiopulmonary bypass with blocking the aorta for 1 hour and then opening the aorta for 15 minutes plus 1 m L physiological saline injection via the femoral vein), experiment group(cardiopulmonary bypass with blocking the aorta for 1 hour and then opening the aorta for 15 minutes plus femoral vein injection of 1 m L physiological saline containing 2×10~7 h AMSCs). Arterial blood samples of 2 m L were taken to calculate oxygenation index and respiratory index before cardiopulmonary bypass(T1), 15 minutes(T2), 1 hour(T3), 2 hours(T4), 3 hours(T5) after opening the aorta. 8 m L intravenous blood samples were taken to detect the serum tumor necrosis factor α, matrix metalloproteinase-9, interleukin-8 and interleukin-10 by ELISA. Meanwhile, western blot assay was used to detect the expression of nuclear factor-κB in lung tissues, and histopathological changes of lung tissues observed under optical microscope. RESULTS AND CONCLUSION: Compared with the control group, the oxygenation index was significantly increased in the experimental group at 2 and 3 hours after transplantation, and the respiratory index was remarkably decreased at 1, 2, 3 hours after transplantation. Compared with the control group, the levels of tumor necrosis factor α, matrix metalloproteinase-9 and interleukin-8 were significantly decreased in the experimental group, but the level of interleukin-10 was increased in the experimental group. Compared with the blank group, the expression of nuclear factor-κB was significantly strengthened in the control group at 3 hours after opening the aorta, but it was decreased significantly after intravenous injection of h AMSCs. After opening the aorta, the pathological changes in the experimental group were remarkably milder than those in the control group, and a small amount of red blood cells and neutrophils exuded, with telangiectasia, and congestion, were found in the experimental group. These findings indicate that h AMSCs transplantation can mitigate ischemia-reperfusioninduced acute lung injury after cardiopulmonary bypass by adjusting the level of nuclear factor-κB in lung tissues and release of tumor necrosis factor α, matrix metalloproteinase-9, interleukin-8 and interleukin-10.更多还原