Autologous induced pluripotent stem cell transplantation for acute lung injury in rats

doi:10.3969/j.issn.2095-4344.1608

Abstract

Abstract:

BACKGROUND: Embryonic stem cells and mesenchymal stem cells are two main cell sources for stem cell transplantation in the treatment of acute lung injury. There are few reports on the study of autologous induced pluripotent stem cells in the treatment of acute lung injury.
OBJECTIVE: To investigate the possibility of induced pluripotent stem cells derived from autologous dermal fibroblasts injected through the caudal vein in the treatment of acute lung injury in rats.
METHODS: Twenty-four Sprague-Dawley rats (provided by Beijing Vital River Laboratory Animal Technology Co., Ltd.) were randomly divided into three groups. The control group was intraperitoneally injected with normal saline, and the model group and the experimental group were intraperitoneally injected with lipopolysaccharide to make acute lung injury models in rats. At 24 hours after modeling, phosphate buffer solution was injected into the tail vein of the rats in the control and model groups, while the rats in the experimental group were given induced pluripotent stem cell suspension by the tail vein. The changes of lung tissue morphology, lung wet/dry weight ratio, pathological injury score, serum interleukin 1beta, interleukin 6 and tumor necrosis factor alpha levels were observed at 7 days after treatment.
RESULTS AND CONCLUSION: (1) At 7 days after treatment, pulmonary interstitial edema, alveolar septum thickening, inflammatory cell infiltration, capillary congestion, irregular alveolar morphology, and exudate in the alveolar cavity were significantly improved in the experimental group. (2) At 7 days after treatment, the wet/dry weight ratio of lung tissue in the model and experimental groups was significantly higher than that in the control group, but the wet/dry weight ratio of lung tissue and pathological injury score in the experimental group were significantly lower than those in the model group (P < 0.01).(3) At 7 days after treatment, the levels of serum interleukin 1beta, interleukin 6 and tumor necrosis factor alpha were ranked as follows: model group > experimental group > control group, and there were significant differences between groups (P < 0.01). To conclude, the transplantation of induced pluripotent stem cells derived from autologous dermal fibroblasts can effectively alleviate acute lung injury and reduce serum inflammatory factor levels in rats.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Acute Lung Injury, Induced Pluripotent Stem Cells, Transplantation, Autologous, Tissue Engineering

CLC Number:

Liu Shenggang, Yang Hongzhong, He Baimei. Autologous induced pluripotent stem cell transplantation for acute lung injury in rats[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(9): 1403-1409.

Figures/Tables 2

2.2 大鼠诱导性多能干细胞的鉴定结果实时荧光定量PCR检测诱导性多能干细胞的多能性，结果显示诱导性多能干细胞表达Oct4、Nanog、SSEA-4和Sox2等多能性基因，见图2A。免疫细胞化学检测诱导性多能干细胞中多能性基因的蛋白表达，结果显示诱导性多能干细胞中Oct3/4、Nanog、SSEA-4和Sox2蛋白呈阳性表达，见图2B。 2.3 实验动物数量分析 24只SD大鼠，随机分为对照组、模型组和实验组，每组8只，整个实验过程顺利，未发生大鼠死亡和造模失败，24只大鼠的实验数据全部进入结果分析。为了观察尾静脉注射诱导性多能干细胞是否影响大鼠存活，该研究将实验大鼠存活能力的观察时间延长为6个月，期间24只大鼠摄食等活动正常，无死亡。 2.4 各组大鼠肺组织湿/干质量比治疗7 d后，模型组和实验组肺组织湿/干质量比明显高于对照组，差异有显著性意义(P < 0.01)，实验组肺组织湿/干质量比明显低于模型组，差异有显著性意义(P < 0.01)，见图3。 2.5 各组大鼠肺组织形态学变化与病理学评分 2.5.1 各组大鼠肺组织形态学变化治疗后肺组织大体形态显示，对照组大鼠肺呈粉红色、表面光滑、无出血点和渗出，见图4A；模型组大鼠肺呈红色、表面粗糙、边缘不规则、有出血和渗出，见图4B；实验组大鼠肺颜色变浅、无出血，水肿和渗出较模型组减轻，见图4C。治疗后肺组织光镜结构显示，对照组大鼠肺组织结构无明显变化，见图4D；模型组大鼠肺组织结构损伤程度加重，肺间质水肿明显，肺泡隔弥漫性增厚，炎细胞和红细胞更多，肺泡形态不规则，大量肺泡消失，肺泡腔内渗出物增多，见图4E；实验组大鼠肺组织损伤较治疗前和模型组有明显改善，肺水肿减轻，浸润的白细胞和红细胞明显减少，部分肺泡隔变薄，肺泡形态仍不规则，肺泡腔内渗出物减少，见图4F。 2.5.2 各组大鼠肺组织病理损伤程度评分治疗前，模型组和实验组病理损伤评分明显高于对照组，差异有显著性意义(P < 0.01)，模型组和实验组病理损伤评分比较，差异无显著性意义(P > 0.05)；治疗后，模型组和实验组病理损伤评分明显高于对照组，差异有显著性意义(P < 0.01)，实验组病理损伤评分明显低于模型组，差异有显著性意义(P < 0.01)；对照组治疗前后病理损伤评分比较，差异无显著性意义(P > 0.05)，模型组治疗后病理损伤评分高于治疗前，差异无显著性意义(P > 0.05)，实验组治疗后病理损伤评分明显低于治疗前，差异有显著性意义(P < 0.01)，见图5。 2.6 各组大鼠治疗前后血清白细胞介素6、白细胞介素1β 和肿瘤坏死因子α水平比较治疗前，模型组和实验组血清3种炎症因子水平明显高于对照组，差异有显著性意义(P < 0.01)，模型组和实验组血清3种炎症因子水平比较，差异无显著性意义(P > 0.05)；治疗后，模型组和实验组血清3种炎症因子水平明显高于对照组，差异有显著性意义(P < 0.01)，实验组血清3种炎症因子水平明显低于模型组，差异有显著性意义(P < 0.01)；对照组治疗前后血清3种炎症因子水平比较，差异无显著性意义(P > 0.05)，模型组治疗后血清3种炎症因子水平高于治疗前，但差异无显著性意义(P > 0.05)，实验组治疗后血清3种炎症因子水平明显低于治疗前，差异有显著性意义(P < 0.01)，见图6。"

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