Liu Hong, Ding Yingwei, Hou Yuehui, Zhao Guangju, Lu Yang, Chen Xiao, Cai Qiqi, Hong Guangliang, Qiu Qiaomeng, Lu Zhongqiu
Emergency Department, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2016 Jan;34(1):1-7. doi: 10.3760/cma.j.issn.1001-9391.2016.01.001.
To explore the possible mechanism and protective effect of BMSCs (bone mesenchymal stem cells) carrying superoxide dismutase (SOD) gene on mice with paraquat-induced acute lung injury.
To establish the cell line of BMSCs bringing SOD gene, lentiviral vector bringing SOD gene was built and co-cultured with BMSCs. A total of 100 BALB/c mice were randomly divided into five groups, namely Control group, poisoning group (PQ group) , BMSCs therapy group (BMSC group) , BMSCs-Cherry therapy group (BMSC-Cherry group) , BMSCs-SOD therapy group (BMSC-SOD group) . PQ poisoning model was produced by stomach lavaged once with 1 ml of 25 mg/kg PQ solution, and the equal volume of normal saline (NS) was given to Control group mice instead of PQ. The corresponding BMSCs therapy cell lines were delivered to mice through the tail vein of mice 4h after PQ treatment.Five mice of each group were sacrificed 3 d, 7 d, 14 d and 21 days after corresponding BMSCs therapy cell lines administration, and lung tissues of mice were taken to make sections for histological analysis. The serum levels of glutathione (GSH) , malondialdehyde (MDA) , SOD, and the levels of transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α) in lung tissue were determined. The level of SOD was assayed by Westen-blot.
Compared with Control group, the early (3 days) levels of SOD protein in lung tissue of PQ group obviously decreased, and the late (21 days) levels of SOD obviously increased, while in therapy groups, that was higher than that in PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) . Compared with Control group, the levels of plasma GSH and SOD of PQ group and each therapy group wae significantly lower than those in Control group, while in therapy groups, those were higher than those of PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) .Compared with Control group, the level of plasma MDA, TNF-α and TGF-β in PQ group and therapy groups were significantly higher, while in therapy groups, that was lower than that in PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) . Lung biopsy showed that, the degree of lung tissue damage in each therapy group obviously reduced.
SOD is the key factor of the removal of reactive oxygen species (ROS) in cells, that can obviously inhibit the oxidative stress damage and the apoptosis induced by PQ, thus significantly increasing alveolar epithelial cell ability to fight outside harmful environment.
探讨携带超氧化物歧化酶(SOD)基因的骨髓间充质干细胞(BMSCs)对百草枯诱导的急性肺损伤小鼠的可能机制及保护作用。
构建携带SOD基因的慢病毒载体并与BMSCs共培养,建立携带SOD基因的BMSCs细胞系。将100只BALB/c小鼠随机分为五组,即对照组、中毒组(PQ组)、BMSCs治疗组(BMSC组)、BMSCs-Cherry治疗组(BMSC-Cherry组)、BMSCs-SOD治疗组(BMSC-SOD组)。用1 ml 25 mg/kg的PQ溶液灌胃一次制备PQ中毒模型,对照组小鼠给予等体积的生理盐水而非PQ。PQ处理4小时后,通过小鼠尾静脉将相应的BMSCs治疗细胞系注入小鼠体内。在给予相应的BMSCs治疗细胞系后3天、7天、14天和21天,每组处死5只小鼠,取小鼠肺组织制作切片进行组织学分析。测定血清中谷胱甘肽(GSH)、丙二醛(MDA)、SOD水平以及肺组织中转化生长因子-β(TGF-β)和肿瘤坏死因子-α(TNF-α)水平。通过蛋白质免疫印迹法检测SOD水平。
与对照组相比,PQ组肺组织中SOD蛋白早期(3天)水平明显降低,晚期(21天)水平明显升高,而治疗组高于PQ组,其中BMSCs-SOD组最明显(均P<0.05)。与对照组相比,PQ组及各治疗组血浆GSH和SOD水平均显著低于对照组,而治疗组高于PQ组,其中BMSCs-SOD组最明显(均P<0.05)。与对照组相比,PQ组及治疗组血浆MDA、TNF-α和TGF-β水平均显著升高,而治疗组低于PQ组,其中BMSCs-SOD组最明显(均P<0.05)。肺活检显示,各治疗组肺组织损伤程度明显减轻。
SOD是细胞内清除活性氧(ROS)的关键因素,可明显抑制PQ诱导的氧化应激损伤和细胞凋亡,从而显著提高肺泡上皮细胞抵御外界有害环境的能力。