Mesenchymal stem cells homing to rat lung emphysema followed by differentiation into pulmonary vascular endothelial cells

doi:10.3969/j.issn.2095-4344.2017.29.02

Abstract

Abstract:

BACKGROUND: Mesenchymal stem cells have unique homing, immunoregulation and anti-inflammatory properties. After intravenous injection, mesenchymal stem cells can home to the damaged target organs and tissues, and function to repair damaged tissues.
OBJECTIVE: To observe the pathological changes of lung tissues in rats with emphysema after mesenchymal stem cells transplantation.
METHODS: Twenty-four female Wistar rats were randomly divided into experimental group, control group and healthy control group. In the first two groups, the model of pulmonary emphysema was established by the method of dropping porcine pancreatic elastase. BrdU-labeled mesenchymal stem cells were injected into the tail vein of the rats in the experimental group, and PBS was injected in the control group. After 14 days, the pathological changes of lung tissues were observed. Tumor necrosis factor-α level, alveolar wall apoptotic index, anti-CD34 and anti-BrdU immunohistochemical changes in the bronchoalveolar lavage fluid were detected.
RESULTS AND CONCLUSION: Compared with the healthy control group, the tumor necrosis factor-α level and apoptotic index of alveolar wall cells increased (P < 0.01), and the relative areas of anti-Brdu and anti-CD34 decreased (P < 0.01) in the control group. Compared with the control group, the level of tumor necrosis factor-α and apoptotic index of alveolar wall cells decreased (P < 0.01), and the relative area of anti-Brdu and anti-CD34 increased (P < 0.01) in the experimental group. The histopathological findings showed that both the control group and the experimental group showed emphysema-like changes, but these changes were milder in the experimental group than the control group. To conclude, mesenchymal stem cells can inhibit inflammatory response and apoptosis in experimental emphysema, improve the pathological changes of the lung, and moreover, bone marrow mesenchymal stem cells can differentiate into lung vascular endothelial cells.

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: Cell transplantation, Stem Cells, Mesenchymal stem cells, Pulmonary Emphysema, Bone marrow, Tissue Engineering

CLC Number:

R394.2

Zhao Xiao-jian, Lu Cai-ping, Chu Wei-wei, Zhang Ya-xiao, Zhang Bing, Zhen Qiang, Wang Ren-feng, Liu Jia-bao. Mesenchymal stem cells homing to rat lung emphysema followed by differentiation into pulmonary vascular endothelial cells[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(29): 4728-4733.

Figures/Tables 1

2.1 培养的骨髓间充质干细胞形态及鉴定细胞悬液中的间充质干细胞悬浮状态，呈圆形，24 h开始贴壁，72 h贴壁的细胞呈梭形、三角形、多角形，胞体饱满，可见2-4个突起，长短不一，胞浆半透明，胞核圆，核仁明显。培养7 d胞体增大，逐渐以梭形为主，细胞间连接更为紧密。传代细胞生长速度加快，更密集，汇流处细胞形态均一，多长梭形，呈旋涡状或平行排列生长，光性强。培养10-14 d细胞80%融合，见图1A，流式细胞仪检测细胞表面CD44表达率为85.32%，CD90表达率为91.45%，CD71表达率为69.34%，见图1B，C，CD45表达率为2.05%[10]。Brdu标记的骨髓间充质干细胞，见图1D。 2.2 肺气肿模型造模情况实验组与对照组大鼠均造模成功。 2.3 各组大鼠肺泡灌洗液细胞总数和细胞分类计数与健康对照组比较，对照组大鼠肺泡灌洗液中细胞总数、巨噬细胞、中性粒细胞、淋巴细胞百分比明显升高(P < 0.01)；与对照组相比较，实验组大鼠肺泡灌洗液中细胞总数明显降低(P < 0.01)，细胞分类计数无明显差异，见表1。 2.4 各组大鼠肺泡灌洗中肿瘤坏死因子α水平检测与健康对照组比较，对照组大鼠肺泡灌洗中肿瘤坏死因子升高(P < 0.01)，表明肺气肿大鼠肺内炎症反应明显；与对照组比较，实验组肿瘤坏死因子水平降低(P < 0.01)，表明间充质干细胞明显降低肺内炎症反应，见表2。 2.5 各组大鼠肺组织形态学观察及测定结果各组肺组织病理变化显示，对照组及实验组肺组织均呈现肺气肿样改变，但实验组较对照组为轻，见图2。对照组和实验组肺组织平均内衬间隔及平均肺泡面积明显高于健康对照组(P < 0.01)，平均肺泡数明显低于健康对照组(P < 0.01)；对照组肺组织平均内衬间隔及平均肺泡面积明显高于实验组(P < 0.01)，平均肺泡数明显低于实验组(P < 0.01)，见表3。 2.6 各组大鼠肺组织肺泡壁凋亡细胞检测结果 TUNEL结果显示凋亡细胞的核染色质浓集、呈深棕褐色。对照组、实验组肺泡壁细胞凋亡数量增多，凋亡指数值显著高于健康对照组(P < 0.01)，实验组肺泡壁细胞凋亡数量及凋亡指数值显著低于对照组(P < 0.01)，见表2。 2.7 各组大鼠肺组织免疫组织化学检测结果见图3。健康对照组、对照组、实验组抗Brdu及抗CD34相对面积分别为(5.63±2.01)%、(0.86±0.49)%、(2.64±1.43)%，3组间两两比较差异均有显著性意义(P < 0.01)。"

References

[1] Lea SR,Metcalfe HJ,Plumb J,et al.Neutral sphingomyelinase-2, acid sphingomyelinase, and ceramide levels in COPD patients compared to controls.Int J Chron Obstruct Pulmon Dis.2016;11:2139-2147.

[2] Khakban A,Sin DD,FitzGerald JM,et al.The Projected Epidemic of COPD Hospitalizations Over the Next 15 Years: A Population Based Perspective.Am J Respir Crit Care Med. 2017;195(3):287-291.

[3] Zhao Y,Cui A,Wang F,et al.Characteristics of pulmonary inflammation in combined pulmonary fibrosis and emphysema.Chin Med J(Engl).2012;125(17):3015-3021.

[4] Tanabe N,Muro S,Tanaka S,et al.Emphysema distribution and annual changes in pulmonary function in male patients with chronic obstructive pulmonary disease.Respir Res.2012; 13:31.

[5] Li BP,Zhao XJ,Song YM,et al.Restoration of pathological changes of emphysema by angiogenesis factors: experiment of rats.Zhonghua Yi Xue Za Zhi.2007;87(33):2365-2368.

[6] Cyranoski D.Stem cells in Texas: Cowboy culture.Nature. 2013;494(7436):166-168.

[7] Vincent LG,Engler AJ.Stem cell differentiation: Post-degradation forces kick in. Nat Mater.2013;12(5): 384-386.

[8] Román F,Urra C,Porras O,et al.Real-Time H2O2 Measurements in Bone Marrow Mesenchymal Stem Cells (MSCs) Show Increased Antioxidant Capacity in Cells from Osteoporotic Women.J Cell Biochem.2017;118(3):585-593.

[9] 李宝平,赵晓建,宋永明,等.血管生成因子修复肺气肿病变的实验研究[J].中华医学杂志,2007,87(33):2365-2368.

[10] Jacobs SA,Roobrouck VD,Verfaillie CM,et al.Immunological characteristics of human mesenchymal stem cells and multipotent adult progenitor cells.Immunol Cell Biol. 2013; 91(1):32-39.

[11] Kennedy GJ.Behavioral Interventions for Patients with Major Depression and Severe COPD.Am J Geriatr Psychiatry. 2016;24(11):975-976.

[12] Bøgelund M,Hagelund L,Asmussen MB.COPD-treating Nurses' Preferences for Inhaler Attributes - a Discrete Choice Experiment.Curr Med Res Opin.2017;33(1):71-75.

[13] Kobayashi S,Hanagama M,Yamanda S,et al.Inflammatory biomarkers in asthma-COPD overlap syndrome.Int J Chron Obstruct Pulmon Dis.2016;11:2117-2123.

[14] Nishio M,Nakane K,Tanaka Y.Application of the homology method for quantification of low-attenuation lung region inpatients with and without COPD.Int J Chron Obstruct Pulmon Dis.2016;11:2125-2137.

[15] Morris NR,Walsh J,Adams L,et al.Exercise training in COPD: What is it about intensity. Respirology.2016;21(7):1185-1192.

[16] Erdal M,Johannessen A,Eagan TMet al.Incidence of utilization- and symptom-defined COPD exacerbations in hospital- and population-recruited patients.Int J Chron Obstruct Pulmon Dis.2016;11:2099-2108.

[17] Miravitlles M,Chapman KR,Chuecos F,et al.The efficacy of aclidinium/formoterol on lung function and symptoms in patients with COPD categorized by symptom status: a pooled analysis.Int J Chron Obstruct Pulmon Dis. 2016;11: 2041-2053.

[18] Choudhury G,MacNee W.Role of Inflammation and Oxidative Stress in the Pathology of Ageing in COPD: Potential Therapeutic Interventions.COPD.2017;14(1):122-135.

[19] ]Malih S,Saidijam M,Mansouri K,et al.Promigratory and proangiogenic effects of AdipoRon on bone marrow-derived mesenchymal stem cells: an in vitro study.Biotechnol Lett. 2017;39(1):39-44.

[20] Guo Q,Chen Y,Guo L,et al.miR-23a/b regulates the balance between osteoblast and adipocyte differentiation in bone marrow mesenchymal stem cells.Bone Res.2016;4:16022.

[21] Ren M,Wang T,Huang L,et al.Role of VR1 in the differentiation of bone marrow-derived mesenchymal stem cells into cardiomyocytes associated with Wnt/β-catenin signaling. Cardiovasc Ther.2016;34(6):482-488.

[22] Chehelcheraghi F,Eimani H,Homayoonsadraie S,et al.Effects of Acellular Amniotic Membrane Matrix and Bone Marrow-Derived Mesenchymal Stem Cells in Improving Random Skin Flap Survival in Rats.Iran Red Crescent Med J.2016;18(6):e25588.

[23] Hao Y,Geng L.Administration of bone marrow mesenchymal stem cells attenuates inflammation of rats with sepsis.Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi.2016; 32(9):1158-1163.

[24] Pérez-Serrano RM,González-Dávalos ML,Lozano-Flores C,et al.PPAR Agonists Promote the Differentiation of Porcine Bone Marrow Mesenchymal Stem Cells into the Adipogenic and Myogenic Lineages.Cells Tissues Organs.2016.[Epub ahead of print]

[25] Zhang J,Shao Y,He D,et al.Evidence that bone marrow-derived mesenchymal stem cells reduce epithelial permeability following phosgene-induced acute lung injury via activation of wnt3a protein-induced canonical wnt/β-catenin signaling.Inhal Toxicol.2016;28(12):572-579.

[26] Chen J,Ma G,Liu W,et al.The influence of the sensory neurotransmitter calcitonin gene-related peptide on bone marrow mesenchymal stem cells from ovariectomized rats.J Bone Miner Metab.2016.[Epub ahead of print]

[27] Cao J,Hou S,Ding H,et al.In Vivo Tracking of Systemically Administered Allogeneic Bone Marrow Mesenchymal Stem Cells in Normal Rats through Bioluminescence Imaging.Stem Cells Int.2016;2016:3970942.

[28] Gong M,Antony S,Sakurai R,et al.Bone marrow mesenchymal stem cells of the intrauterine growth restricted rat offspring exhibit enhanced adipogenic phenotype. Int J Obes (Lond).2016;40(11):1768-1775.

[29] Roger Y,Schäck LM,Koroleva A,et al.Grid-like surface structures in thermoplastic polyurethane induce anti-inflammatory and anti-fibrotic processes in bone marrow-derived mesenchymal stem cells.Colloids Surf B Biointerfaces.2016;148: 104-115.

[30] Fu WM,Yang L,Wang BJ,et al.Porous tantalum seeded with bone marrow mesenchymal stem cells attenuates steroid-associated osteonecrosis.Eur Rev Med Pharmacol Sci.2016; 20(16):3490-3499.

[31] Dos SGG,Batool S,Hastreiter A,et al.The influence of protein malnutrition on biological and immunomodulatory aspects of bone marrow mesenchymal stem cells.Clin Nutr. 2016.pii: S0261-5614(16)30202-3.doi:10.1016/j.clnu.2016.08.005.[Epub ahead of print]