Liu F-B, Lin Q, Liu Z-W
Department of Thoracic Surgery, Shanghai General Hospital, Shanghai Jiaotong University, Shanghai, China.
Eur Rev Med Pharmacol Sci. 2016 Mar;20(5):969-82.
We sought to determine whether normal human umbilical cord mesenchymal stem cells and apoptotic human umbilical cord mesenchymal stem cells play any role in the lung repair following bleomycin-induced lung injury in rat models.
Umbilical cord mesenchymal stem cells were obtained from the umbilical cord following caesarian section from healthy normal babies. Plasmin deprivation method was used for culture of human umbilical cord mesenchymal stem cells and flow cytometry was used to identify cell surface antigen and activity of stem cells and apoptosis. The animal model of acute lung injury was established by a one-off intratracheal instillation of bleomycin (BLM) (5 mg/kg) and then normal stem cells and apoptotic stem cells were separately injected. Alveolar lavage fluid and lung tissue were collected for further analysis prior to the injury and at days 3, 7, 14 after administration of BLM. The number of neutrophils in the broncho alveolar lavage fluid (BALF) was counted; Bicinchoninic Acid (BCA) method was used for estimation of total protein content in alveolar lavage fluid; biochemical assay was used for estimation of myeloperoxidase (MPO) activity; hematoxylin and eosin (HE) staining of lung tissue was used for histopathology analysis; reverse transcription-polymerase chain reaction (RT-PCR) assay was used for the determination of interferon-gamma (INF-γ) and mRNA changes of interleukin-4 (IL-4) in lung tissue. Enzyme-linked immunosorbent assay (ELISA) was used for the determination of cytokines TNF-α in the lung tissue.
Apoptotic human umbilical cord mesenchymal stem cells were more effective in reducing lung neutrophil infiltration and total protein leakage in rat models of acute lung injury (ALI). There was also an improvement in the degree of vascular permeability, reduction in the level of proinflammatory cytokines, INF-γ gene level and boost in anti-inflammatory cytokine IL-4 levels which also helps in more effectively reducing the degree of injury in ALI.
Human umbilical cord mesenchymal stem cell transplantation may have a bright future in the clinical setting for the treatment of ALI/acute respiratory distress syndrome (ARDS). Apoptotic human umbilical cord mesenchymal stem cells may have more effective than normal human umbilical cord mesenchymal stem cells in the treatment of acute lung injury.
我们试图确定正常人类脐带间充质干细胞和凋亡的人类脐带间充质干细胞在博来霉素诱导的大鼠肺损伤后的肺修复中是否发挥任何作用。
从健康正常婴儿剖宫产术后的脐带中获取脐带间充质干细胞。采用纤溶酶剥夺法培养人类脐带间充质干细胞，并用流式细胞术鉴定细胞表面抗原、干细胞活性及凋亡情况。通过一次性气管内滴注博来霉素（BLM）（5mg/kg）建立急性肺损伤动物模型，然后分别注射正常干细胞和凋亡干细胞。在损伤前以及给予BLM后第3、7、14天收集肺泡灌洗液和肺组织进行进一步分析。计数支气管肺泡灌洗液（BALF）中的中性粒细胞数量；采用双辛可宁酸（BCA）法估算肺泡灌洗液中的总蛋白含量；采用生化分析法估算髓过氧化物酶（MPO）活性；对肺组织进行苏木精-伊红（HE）染色用于组织病理学分析；采用逆转录-聚合酶链反应（RT-PCR）分析法测定肺组织中干扰素-γ（INF-γ）和白细胞介素-4（IL-4）的mRNA变化。采用酶联免疫吸附测定（ELISA）法测定肺组织中的细胞因子肿瘤坏死因子-α（TNF-α）。
在急性肺损伤（ALI）大鼠模型中，凋亡的人类脐带间充质干细胞在减少肺中性粒细胞浸润和总蛋白渗漏方面更有效。血管通透性程度也有所改善，促炎细胞因子水平降低，INF-γ基因水平降低，抗炎细胞因子IL-4水平升高，这也有助于更有效地降低ALI的损伤程度。
人类脐带间充质干细胞移植在急性肺损伤/急性呼吸窘迫综合征（ARDS）的临床治疗中可能具有光明的前景。凋亡的人类脐带间充质干细胞在治疗急性肺损伤方面可能比正常人类脐带间充质干细胞更有效。
