Zhang Xiaomiao, Gao Fengying, Yan Yunqi, Ruan Zheng, Liu Zhenwei
Department of Thoracic Surgery, First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, China.
Cell Biochem Funct. 2015 Apr;33(3):113-20. doi: 10.1002/cbf.3092. Epub 2015 Mar 10.
Acute lung ischemia-reperfusion injury (ALIRI) is a serious disease that seriously affects human's life. In this study, we aimed to explore a more effective treatment method by combining human umbilical cord mesenchymal stem cells (HUMSCs) and angiotensin-converting enzyme 2 (ACE2) for ALIRI. Fifty rats were firstly divided into five groups, namely sham surgery group (sham) and four model groups (model, ACE2, HUMSCs and HUMSCs + ACE2) that were reperfused with 0.1 ml physiological saline (PS), 0.1 ml PS containing 1 × 10(6) lentiviral-ACE2/HUMSCs/ACE2 + UMSCs, respectively. Quantitative reverse transcription-PCR (qRT-PCR) and western blot assays were then conducted to detect the messenger RNA (mRNA) and protein levels of inflammatory cytokines [intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), tumour necrosis factor α (TNF-α), nuclear factor κB (NF-κB), platelet-derived growth factor (PDGF) and angiotensin II (Ang II)], antioxidant proteins [NAD(P)H quinone oxidoreductase 1 (NQO1), heme oxygenase 1 (HO-1)], DNA damage and apoptotic indicators [BCL2-associated X (Bax), cleaved caspase-3 (C-Csp 3), cleaved-poly(ADP-ribose) polymerase (C-PARP), Y-H2AX], anti-apoptotic indicator (Bcl-2) and smooth muscle cell proliferation indicator [connexin 43 (Cx43)]. According to the qRT-PCR and western results, the mRNA and protein expression levels of ICAM-1, VCAM-1, TNF-α, NF-κB, PDGF, Bax, C-Csp 3, C-PARP and Y-H2AX were significantly higher in model group than those in sham group and they were significantly reduced by HUMSCs or ACE2 treatment (P < 0.05). On the contrary, Bcl-2 showed an opposite expression trend with the previous proteins. The mRNA and protein levels of NQO1 and HO-1 were sequentially increased in sham, model, ACE2, HUMSCs and HUMSCs + ACE2 groups. Besides, HUMSCs combined with ACE2 exhibited a better inhibition effect on ALIRI than HUMSCs or ACE2 alone (P < 0.05). In summary, HUMSCs combined with ACE2 was demonstrated to have the best therapeutic effect on ALIRI through anti-inflammation, oxidative stress and anti-apoptotic processes.
急性肺缺血再灌注损伤（ALIRI）是一种严重影响人类生命的疾病。在本研究中，我们旨在探索一种更有效的治疗方法，即将人脐带间充质干细胞（HUMSCs）与血管紧张素转换酶2（ACE2）联合用于治疗ALIRI。首先将50只大鼠分为五组，即假手术组（sham）和四个模型组（模型组、ACE2组、HUMSCs组和HUMSCs+ACE2组），分别用0.1 ml生理盐水（PS）、0.1 ml含1×10(6)慢病毒-ACE2/HUMSCs/ACE2+UMSCs的PS进行再灌注。然后进行定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹分析，以检测炎症细胞因子[细胞间黏附分子1（ICAM-1）、血管细胞黏附分子1（VCAM-1）、肿瘤坏死因子α（TNF-α）、核因子κB（NF-κB）、血小板衍生生长因子（PDGF）和血管紧张素II（Ang II）]、抗氧化蛋白[NAD(P)H醌氧化还原酶1（NQO1）、血红素加氧酶1（HO-1）]、DNA损伤和凋亡指标[BCL2相关X蛋白（Bax）、裂解的半胱天冬酶-3（C-Csp 3）、裂解的聚（ADP-核糖）聚合酶（C-PARP）、Y-H2AX]、抗凋亡指标（Bcl-2）和平滑肌细胞增殖指标[连接蛋白43（Cx43）]的信使核糖核酸（mRNA）和蛋白质水平。根据qRT-PCR和蛋白质印迹结果，模型组中ICAM-1、VCAM-1、TNF-α、NF-κB、PDGF、Bax、C-Csp 3、C-PARP和Y-H2AX的mRNA和蛋白质表达水平显著高于假手术组，而HUMSCs或ACE2治疗可使其显著降低（P<0.05）。相反，Bcl-2与上述蛋白质呈现相反的表达趋势。NQO1和HO-1的mRNA和蛋白质水平在假手术组、模型组、ACE2组、HUMSCs组和HUMSCs+ACE2组中依次升高。此外，与单独使用HUMSCs或ACE2相比，HUMSCs与ACE2联合使用对ALIRI的抑制作用更好（P<0.05）。综上所述，通过抗炎、氧化应激和抗凋亡过程，证明HUMSCs与ACE2联合使用对ALIRI具有最佳治疗效果。
