Tanreqing injection combined with human amniotic mesenchymal stem cell transplantation exerts protective effect against blood transfusion related acute lung injury

doi:10.3969/j.issn.2095-4344.2017.21.013

Abstract

Abstract:

BACKGROUND: With the improvement of cellular technology, stem cell transplantation has been involved in various diseases, such as transfusion-related acute lung injury (TRALI).

OBJECTIVE: To investigate the protective effect of Tanreqing injection combined with human amniotic mesenchymal stem cells (hAMSCs) transplantation in a mouse model of TRALI.
METHODS: A total of 80 BALB/C male mice were divided into control group, model group, Tanreqing injection group, hAMSCs transplantation group and combined group (Tanreqing injection+hAMSCs transplantation group). Animal models of TRALI were made in all groups except the control group. Mouse lung tissue pathology and wet/dry ratio were observed and calculated 24 hours after treatment. ELISA was used to measure interleukin-10, interleukin-1β, and tumor necrosis factor-α levels in bronchoalveolar lavage fluid. Expression of keratinocyte growth factor-1 mRNA and protein was detected by RT-PCR and western blot, respectively. Apoptosis in the lung tissue was detected by TUNEL method.
RESULTS AND CONCLUSION: (1) Thickened alveolar septum, alveolar fibrin infiltration, alveolar hemorrhage, thickened bronchial wall, and increased lung wet/dry ratio were observed in the model group, and these pathological changes were slightly milder in the Tanreqing injection group and hAMSCs transplantation group. Compared with the model group, the lung wet/dry ratio was significantly lower in the Tanreqing injection group and hAMSCs transplantation group (P < 0.05), and very significantly lower in the combined group (P < 0.01). (2) Compared with the model group, the levels of interleukin-10, interleukin-1β, and tumor necrosis factor-α in the bronchoalveolar lavage fluid were significantly lower in the Tanreqing injection group and hAMSCs transplantation group (P < 0.05), and very significantly lower in the combined group (P < 0.01), but these levels were still higher in the combined group than the control group (P < 0.05). (3) The expression levels of keratinocyte growth factor-1 mRNA and protein were ranked as follows: combined group > Tanreqing injection group and hAMSCs transplantation group > model group, and there were significant differences between them (P < 0.05). (4) The apoptotic index was ranked as follows: combined group < Tanreqing injection group and hAMSCs transplantation group < model group. To conclude, Tanreqing injection combined with hAMSCs transplantation can improve expression of keratinocyte growth factor-1 in the lung tissue of TRALI mice, reduce degree of lung inflammatory reaction and pathologic injury, and reduce the apoptosis in the lung tissue, which plays a protective role against lung injury in mice.

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, Amnion, Mesenchymal Stem Cell Transplantation, Tissue Engineering

CLC Number:

R394.2

Gu Pei-hong. Tanreqing injection combined with human amniotic mesenchymal stem cell transplantation exerts protective effect against blood transfusion related acute lung injury[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(21): 3358-3363.

Figures/Tables 2

2.1 人羊膜间充质干细胞的形态取冻存的人羊膜间充质干细胞进行复苏，48 h后几乎全部细胞贴壁生长且晃动培养瓶细胞不易脱离，与原代培养细胞无明显差异，即大部分细胞具有较为一致的形态，多呈梭形，呈漩涡状或放射状生长(图1)。 2.2 各组小鼠肺组织病理及湿干比变化对照组肺组织结构清晰，无充血、水肿及炎症细胞。其余4组肺组织中可见不同程度的炎症表现，即支气管和肺组织结构紊乱、充血，支气管管壁增厚等(图2)，人羊膜间充质干细胞移植组和痰热清注射液组以上病理变化较模型组略轻，联合治疗组肺组织结构接近于对照组。模型组肺组织湿干质量比高于对照组(P < 0.05)，人羊膜间充质干细胞移植组和痰热清注射液组肺组织湿干质量比低于模型组(P < 0.05)，联合治疗组肺组织湿干质量比明显低于模型组(P < 0.01)，见表1。 2.3 PKH-26标记的人羊膜间充质干细胞分布以PKH-26标记阳性的细胞呈现红色荧光，在人羊膜间充质干细胞移植组和联合治疗组小鼠肺组织中可见人羊膜间充质干细胞均有红色荧光，表明该方法标记敏感性好，标记率高，而其余各组未见阳性细胞出现，见图3。 2.4 各组小鼠支气管肺泡灌洗液中白细胞介素10、白细胞介素1β、肿瘤坏死因子α水平人羊膜间充质干细胞移植组和痰热清注射液组小鼠肺泡灌洗液中白细胞介素10、白细胞介素1β、肿瘤坏死因子α水平均低于模型组(P < 0.05)；联合治疗组白细胞介素10、白细胞介素1β、肿瘤坏死因子α水平明显低于模型组(P < 0.01)，仍高于对照组(P < 0.05)，见表2。 2.5 各组小鼠肺组织KGF-1 mRNA表达与人羊膜间充质干细胞移植组和痰热清注射液组相比，联合治疗组KGF-1 mRNA的表达升高，差异有显著性意义，而人羊膜间充质干细胞移植组和痰热清注射液组KGF-1mRNA的表达高于模型组，差异有显著性意义，见图4。 2.6 各组小鼠肺组织KGF-1蛋白表达联合治疗组KGF-1蛋白的表达高于人羊膜间充质干细胞移植组和痰热清注射液组，而人羊膜间充质干细胞移植组和痰热清注射液组KGF-1蛋白的表达高于模型组，差异均有显著性意义(P < 0.05)，见图5。 2.7 各组小鼠肺组织细胞凋亡情况与对照组比较，模型组凋亡细胞明显增多，人羊膜间充质干细胞移植组和痰热清注射液组凋亡细胞数低于模型组，联合治疗组凋亡细胞较少，见图6。"

References

[1] Land WG. Transfusion-Related Acute Lung Injury: The Work of DAMPs. Transfus Med Hemother. 2013;40(1):3-13.

[2] Popovsky MA. Transfusion-Related Acute Lung Injury: Incidence, Pathogenesis and the Role of Multicomponent Apheresis in Its Prevention. Transfus Med Hemother. 2008; 35(2):76-79.

[3] Toy P, Gajic O, Bacchetti P, et al. Transfusion-related acute lung injury: incidence and risk factors. Blood. 2012;119(7): 1757-1767.

[4] Yost CS, Matthay MA, Gropper MA. Etiology of acute pulmonary edema during liver transplantation : a series of cases with analysis of the edema fluid. Chest. 2001;119(1):219-223.

[5] Gilliss BM, Looney MR. Experimental models of transfusion- related acute lung injury. Transfus Med Rev. 2011;25(1):1-11.

[6] Sayah DM, Looney MR, Toy P. Transfusion reactions: newer concepts on the pathophysiology, incidence, treatment, and prevention of transfusion-related acute lung injury. Crit Care Clin. 2012;28(3):363-372.

[7] Lindenmair A, Hatlapatka T, Kollwig G, et al. Mesenchymal stem or stromal cells from amnion and umbilical cord tissue and their potential for clinical applications. Cells. 2012;1(4): 1061-1088.

[8] 胡泽斌,王立生,崔春萍,等.干细胞临床应用安全性评估报告[J],中国医药生物技术,2013,8(5):349-361.

[9] Kharaziha P, Hellström PM, Noorinayer B, et al. Improvement of liver function in liver cirrhosis patients after autologous mesenchymal stem cell injection: a phase I-II clinical trial. Eur J Gastroenterol Hepatol. 2009;21(10):1199-1205.

[10] 杨华强,张荣环,杜玲,等.脐带间充质干细胞移植治疗脊髓损伤的临床研究[J].现代生物医学进展,2012,12(3):518-521.

[11] Amable PR, Teixeira MV, Carias RB, et al. Protein synthesis and secretion in human mesenchymal cells derived from bone marrow, adipose tissue and Wharton's jelly. Stem Cell Res Ther. 2014;5(2):53.

[12] de Girolamo L, Lucarelli E, Alessandri G, et al. Mesenchymal stem/stromal cells: a new ''cells as drugs'' paradigm. Efficacy and critical aspects in cell therapy. Curr Pharm Des. 2013; 19(13):2459-2473.

[13] 牟乐明,孙占胜,王伯珉,等.骨髓间充质干细胞移植对脊髓损伤大鼠Toll样受体4表达的影响[J].山东大学学报:医学版,2014, 52(1): 37-41.

[14] Sun ZM, Liu HL, Geng LQ, et al. HLA-matched sibling transplantation with G-CSF mobilized PBSCs and BM decreases GVHD in adult patients with severe aplastic anemia. J Hematol Oncol. 2010;3:51.

[15] Saito S, Lin YC, Murayama Y, et al. Human amnion-derived cells as a reliable source of stem cells. Curr Mol Med. 2012; 12(10):1340-1349.

[16] 蒋旭宏,黄小民,何煜舟.痰热清注射液对急性肺损伤大鼠肺组织抗氧化作用的影响[J].中华中医药学刊,2012,30(5): 1043-1045.

[17] 闫龙,黄小民.痰热清注射液对急性肺损伤大鼠保护机制的实验研究[J].中国中医急症,2011,20(4):587-589.

[18] 李丽玮,花紫菱,张惠箴,等.输血相关急性肺损伤大鼠模型建立及肺组织病理研究[J].中国输血杂志,2012,25(11):1121-1124.

[19] 刘晓宇,闫浩,朱清仙,等.标准化粉尘螨疫苗免疫治疗哮喘小鼠肺组织病理变化动态观察[J].热带医学杂志,2011,11(11): 1234-1247.

[20] 赵敏,师霞,邱桐,等. 黄芪对急性肺损伤大鼠模型肺组织中角质细胞生长因子表达的影响[J].中医临床研究,2013,5(5):11-13.

[21] 万巧凤,顾立刚,殷胜骏,等.黄芩苷对 FM1 肺炎小鼠肺组织细胞凋亡FAS / FAS-L 系统的影响[J].中国药理学通报,2011,27(11): 1555-1559.

[22] 王方,张小岗,张静,等.自体骨髓干细胞治疗失代偿期肝硬化患者50例疗效观察[J].实用肝脏病杂志,2010,13(4):272-274.

[23] Ma S, Xie N, Li W, et al. Immunobiology of mesenchymal stem cells. Cell Death Differ. 2014;21(2):216-225.

[24] Cuiffo BG, Karnoub AE. Mesenchymal stem cells in tumor development: emerging roles and concepts. Cell Adh Migr. 2012;6(3):220-230.

[25] Parolini O, Caruso M. Review: Preclinical studies on placenta-derived cells and amniotic membrane: an update. Placenta. 2011;32 Suppl 2:S186-195.

[26] Manochantr S, U-pratya Y, Kheolamai P, et al. Immunosuppressive properties of mesenchymal stromal cells derived from amnion, placenta, Wharton's jelly and umbilical cord. Intern Med J. 2013;43(4):430-439.

[27] 朴正福,张海燕,丁淑芹,等.人羊膜间充质干细胞的生物学特性鉴定[J].国际生物医学工程杂志,2010,33(3):157-162.

[28] 王利梅.痰热清注射液的作用机制及不良反应[J].中医药临床杂志,2015,27(1):110-111.

[29] 祝晨,黄小民.痰热清注射液对内毒素型急性肺损伤大鼠的保护作用[J].中华中医药学刊,2012,30(8):1743-1745.

[30] 梁迪思,张智琳,江桂英,等.不同剂量痰热清注射液对急性肺损伤大鼠白细胞介素-6的影响[J].广东医学,2013,34(8):1170-1172.

[31] 闫龙,来毅.痰热清注射液对急性肺损伤大鼠的肺组织核因子-κB表达的影响[J].中国中医急症,2015,24(1):38-41.

[32] 蒋旭宏,黄小民,何煜舟.痰热清注射液对急性肺损伤大鼠肺组织抗氧化作用的影响[J].中华中医药学刊,2012,30(5):1043-1045.