Galectin-1 secreted by bone marrow mesenchymal stem cells may suppress airway inflammation in an asthmatic mouse

doi:10.3969/j.issn.2095-4344.1767

Abstract

Abstract:

BACKGROUND: Our previous studies have demonstrated that bone marrow mesenchymal stem cells can ameliorate airway inflammation and regulate levels of inflammatory factors. Moreover, galectin-1 secreted by bone marrow mesenchymal stem cells can inhibit the immune function of dendritic cells in vitro. However, the effect of galectin-1 secreted by bone marrow mesenchymal stem cells on asthma is unknown.
OBJECTIVE: To investigate the effect of galectin-1 secreted by bone marrow mesenchymal stem cells on airway inflammation of asthmatic mice.
METHODS: A recombinant lentiviral vector, pLVX-gal-1-shRNA, was constructed for RNA interference of galectin-1 gene and then was transferred into bone marrow mesenchymal stem cells. Forty female BALB/c mice were equally randomized into normal control group, asthmatic group, bone marrow mesenchymal stem cells treatment group (cell treatment group), galectin-1 treatment group and galectin-1 interference group. The number of total inflammatory cells and differential cells in the mouse bronchoalveolar lavage fluid was determined. Furthermore, hematoxylin-eosin staining was used to compare airway inflammation among five groups.
RESULTS AND CONCLUSION: Four short-hairpin RNA sequences targeting mouse galectin-1 mRNA were designed. A real time-quantitative polymerase chain reaction demonstrated that bone marrow mesenchymal stem cells were inhibited most by interference site 249 RNA sequences which were later used in vivo study. Accumulation of inflammation cells, particularly eosinophils, around the airway and in the bronchoalveolar lavage fluid was evidence in asthmatic mice compared to the normal control group. However, bone marrow mesenchymal stem cells engraftment or protein of galectin-1 infusion significantly reduced inflammatory infiltration both in the airway and in the bronchoalveolar lavage fluid. Moreover, eosinophils in the bronchoalveolar lavage fluid and in the airway attenuated dramatically in the cell treatment group and galectin-1 treatment group. However, there was no effect on inflammation accumulation in the bronchoalveolar lavage fluid and airway by infusion of galectin-1-transfected bone marrow mesenchymal stem cells to asthmatic mice. These results indicate that galectin-1 secreted by bone marrow mesenchymal stem cells could alleviate airway inflammation in the asthmatic mouse.

Key words: asthma, bone marrow mesenchymal stem cell, galectin-1, lentiviral interference vector, airway inflammation, inflammatory cells, eosinophils, National Natural Science Foundation of China

CLC Number:

Ge Xiahui, Zhang Guorui, Guan Wenbin, Bai Chong. Galectin-1 secreted by bone marrow mesenchymal stem cells may suppress airway inflammation in an asthmatic mouse[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(25): 3937-3943.

Figures/Tables 5

2.5 肺泡灌洗液细胞计数如图5所示，正常对照组肺泡灌洗液内可见散在的支气管上皮细胞，少许单核细胞及淋巴细胞；哮喘小鼠肺泡灌洗液中可见大量的炎性细胞，以嗜酸性粒细胞为主，单核细胞及中性粒细胞亦较正常对照组增多，细胞总数较正常对照组明显增多；BMSCs组或重组半乳凝素1蛋白组肺泡灌洗液中细胞总数较哮喘模型组明显减少，其中嗜酸性粒细胞及单核细胞较哮喘模型组显著减少，而中性粒细胞较哮喘模型组略有增加，但差异无显著性意义；哮喘小鼠予以gal-1-RNAi-BMSCs移植未见肺泡灌洗液中细胞总数减少，嗜酸性粒细胞及单核细胞也未见减少，因此体内注射干扰半乳凝素1基因的骨髓间充质干细胞抑制了BMSCs对哮喘小鼠的治疗作用。进一步对各组哮喘肺泡灌洗液细胞进行分类及计数，如图6所示，与正常对照组比较，哮喘模型组肺泡灌洗液中嗜酸性粒细胞比例明显增高(P < 0.01)；与哮喘模型组比较，输注重组半乳凝素1蛋白或移植BMSCs使其嗜酸性粒细胞比例明显降低(P < 0.01)，gal-1-RNAi-BMSCs移植并不能减少肺泡灌洗液中嗜酸性粒细胞的比例(P=0.964)。与正常对照组比较，哮喘模型组肺泡灌洗液中单核细胞比例明显增高(P < 0.01)，与哮喘模型组比较，BMSCs组或重组半乳凝素1蛋白组肺泡灌洗液中单核细胞比例明显降低，而gal-1-RNAi-BMSCs移植使哮喘小鼠肺泡灌洗液中单核细胞数略增高(P=0.024)。BMSCs或重组半乳凝素1蛋白移植对中性粒细胞无显著影响。与正常对照组比较，哮喘模型组肺泡灌洗液中细胞总数明显增高(P < 0.01)；与哮喘模型组比较，BMSCs组或重组半乳凝素1蛋白组肺泡灌洗液中细胞总数明显下降(P < 0.01)，gal-1-RNAi-BMSCs移植并不能减少哮喘小鼠肺泡灌洗液中细胞总数(P=0.955)。"

References

[1]Ge X, Bai C, Yang J, et al. Effect of mesenchymal stem cells on inhibiting airway remodeling and airway inflammation in chronic asthma. J Cell Biochem. 2013;114(7):1595-1605.

[2]Ge X, Bai C, Yang J, et al. Intratracheal transplantation of bone marrow-derived mesenchymal stem cells reduced airway inflammation and up-regulated CD4?CD25? regulatory T cells in asthmatic mouse. Cell Biol Int. 2013;37(7):675-686.

[3]戈霞晖,白冲,陈若华,等.骨髓间充质干细胞移植对慢性哮喘小鼠气道炎症及气道重塑的影响[J].中国呼吸与危重监护杂志,2011, 10(1):6-10.

[4]戈霞晖,白冲,陈若华,等.骨髓间充质干细胞对慢性哮喘小鼠血清IL-12和IL-4水平的影响[J].中国组织工程研究与临床康复杂志, 2011,15(6):1030-1034.

[5]Zhang Y, Ge XH, Guo XJ, et al. Bone Marrow Mesenchymal Stem Cells Inhibit the Function of Dendritic Cells by Secreting Galectin-1. Biomed Res Int. 2017;2017:3248605.

[6]沈璐,赖克方,姜华,等. 不同激发方式对小鼠过敏性支气管哮喘模型的影响[J].中华哮喘杂志(电子版),2009,3(6):404-408.

[7]Rabinovich GA, Toscano MA. Turning 'sweet' on immunity: galectin-glycan interactions in immune tolerance and inflammation. Nat Rev Immunol. 2009;9(5):338-352.

[8]Barrionuevo P, Beigier-Bompadre M, Ilarregui JM, et al. A novel function for galectin-1 at the crossroad of innate and adaptive immunity: galectin-1 regulates monocyte/macrophage physiology through a nonapoptotic ERK-dependent pathway. J Immunol. 2007;178(1):436-445.

[9]Moore WC, Meyers DA, Wenzel SE, et al. Identification of asthma phenotypes using cluster analysis in the Severe Asthma Research Program. Am J Respir Crit Care Med. 2010;181(4):315-323.

[10]Gibson PG. Inflammatory phenotypes in adult asthma: clinical applications. Clin Respir J. 2009;3(4):198-206.

[11]Wenzel SE. Asthma: defining of the persistent adult phenotypes. Lancet. 2006;368(9537):804-813.

[12]impson JL, Scott R, Boyle MJ, et al. Inflammatory subtypes in asthma: assessment and identification using induced sputum. Respirology. 2006;11(1):54-61.

[13]Halwani R, Al-Muhsen S, Al-Jahdali H, et al. Role of transforming growth factor-β in airway remodeling in asthma. Am J Respir Cell Mol Biol. 2011;44(2):127-133.

[14]Castro M, Wenzel SE, Bleecker ER, et al. Benralizumab, an anti-interleukin 5 receptor α monoclonal antibody, versus placebo for uncontrolled eosinophilic asthma: a phase 2b randomised dose-ranging study. Lancet Respir Med. 2014; 2(11):879-890.

[15]Lin YD, Fan XL, Zhang H, et al. The genes involved in asthma with the treatment of human embryonic stem cell-derived mesenchymal stem cells. Mol Immunol. 2018;95:47-55.

[16]Lepelletier Y, Lecourt S, Renand A, et al. Galectin-1 and semaphorin-3A are two soluble factors conferring T-cell immunosuppression to bone marrow mesenchymal stem cell. Stem Cells Dev. 2010;19(7):1075-1079.

[17]Sioud M, Mobergslien A, Boudabous A, et al. Mesenchymal stem cell-mediated T cell suppression occurs through secreted galectins. Int J Oncol. 2011;38(2):385-390.

[18]Ge XN, Ha SG, Greenberg YG, et al. Regulation of eosinophilia and allergic airway inflammation by the glycan-binding protein galectin-1. Proc Natl Acad Sci U S A. 2016;113(33):E4837-4846.