Bone marrow mesenchymal stem cells regulate nuclear factor kappaB expression in alveolar macrophages of acute lung injury rats with sepsis

doi:10.3969/j.issn.2095-4344.2015.10.015

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells have a therapeutic effect on acute lung injury, but the mechanism is unclear. If the mechanism is understood, the majority of patients with acute lung injury can obtain a benefit.

OBJECTIVE: To explore the possible mechanism underlying bone marrow mesenchymal stem cells in the treatment of acute lung injury with sepsis in rats.

METHODS: (1) Thirty-six adult Wistar rats were randomly divided into three groups, sham operation group (sham group), sepsis group and bone marrow mesenchymal stem cells group (cell treatment group). In the sepsis and cell treatment groups, animal models of sepsis with acute lung injury were established by cecal ligation and puncture, while in the sham group, the cecum was not ligated and punctured. Then, 1 mL normal saline was injected via the femoral vein in the sepsis and sham groups, and 1 mL bone marrow mesenchymal stem cell suspension (1×10⁹/L) was injected into the cell treatment group. After 6 hours, interleukin 10 and macrophage inflammatory protein-2 levels in serum were measured in the three groups. Lung tissues were taken for pathological observation using hematoxylin-eosin staining. (2) Rat alveolar macrophages were obtained by bronchoalveolar lavage, seeded into 24-well culture plates, and divided into three groups: control group (group A), sepsis model group (group B) and intervention group of bone marrow mesenchymal stem cells (group C). Normal saline, septic plasma, and co-intervention of septic plasma and mesenchymal stem cells were used in the groups A, B, C, respectively. Then, cells in the three groups were cultured in a 5% CO₂ incubator at 37 ℃ for 1 hour. After that, alveolar macrophages were taken to detect whether nuclear factor-κB (P65) protein entered into the nucleus using laser scanning confocal microscopy.

RESULTS AND CONCLUSION: (1) The results of animal experiments showed that compared with the sham group, the macrophage inflammatory protein-2 levels in the sepsis group and cell treatment group were significantly increased (P < 0.05), but the macrophage inflammatory protein-2 level in the cell treatment group was significantly lower than that in the sepsis group (P < 0.05); there were no significant differences in serum interleukin 10 levels among the three groups (P > 0.05); inflammatory cell infiltration, interstitial pulmonary edema and pulmonary hemorrhage existed in the sepsis and cell treatment groups, but these symptoms were significantly reduced in the cell treatment group compared with the sepsis group. (2) Results from cell experiments showed that compared with the group A, in group B and group C, the number of nuclear factor-κB (P65) proteins into the nucleus was significantly higher (P < 0.05), but it was lower in the group C than the group B (P < 0.05). These findings indicate that bone marrow mesenchymal stem cells in acute lung injury with sepsis can regulate nuclear factor-κB (P65) protein of alveolar macrophages into the nucleus, reduce expression of macrophage inflammatory protein-2, and thereby play a protective role in the lung via reducing neutrophil infiltration. Temporarily, this study cannot explain whether bone marrow mesenchymal stem cells have an effect on interleukin 10.

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

全文链接：

Key words: Bone Marrow, Mesenchymal Stem Cell Transplantation, Sepsis, Lung Injury, NF-kappa B, Macrophage Inflammatory Proteins

CLC Number:

R394.2

Zhang Ji-feng, Zhang Zi-qi, Luo Xiao-tian, Hou Lin-yi, Jiang Qin, Lv Jie-ping, Zhang Wen-kai. Bone marrow mesenchymal stem cells regulate nuclear factor kappaB expression in alveolar macrophages of acute lung injury rats with sepsis[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(10): 1556-1561.

Figures/Tables 6

References

[1]Bersten AD, Edibam C, Hunt T, et al. Incidence and mortality of acute lung injury and the acute respiratory distress syndrome in three Australian States. Am J Respir Crit Care Med. 2002;165(4):443-448.
[2]Li G, Malinchoc M, Cartin-Ceba R, et al. Eight-year trend of acute respiratory distress syndrome: a population-based study in Olmsted County, Minnesota. Am J Respir Crit Care Med. 2011;183(1):59-66.
[3]张文凯,吕洁萍,强准,等.肺泡巨噬细胞在脓毒症急性肺损伤中的作用[J].中国药物与临床, 2013,13(7):876-878.
[4]Downey GP, Dong Q, Kruger J, et al. Regulation of neutrophil activation in acute lung injury. Chest. 1999;116(1 Suppl): 46S-54S.
[5]Müller I, Lymperi S, Dazzi F. Mesenchymal stem cell therapy for degenerative inflammatory disorders. Curr Opin Organ Transplant. 2008;13(6):639-644.
[6]李霖,卿国忠,杨靖等.急性肺损伤大鼠肺组织神经导向因子Slit2及Robo4的表达[J].中华急诊医学杂志, 2014,23(3):261-266.
[7]Davin MS.Confocal scanning optical microscopy and its applications for biological specimens. J Cell Science. 1989; 94(2):175-182.
[8]沈华浩,张根生. 从炎症本质看急性肺损伤/急性呼吸窘迫综合征[J].中华急诊医学杂志, 2006,15(11):965-966.
[9]强准,张继峰,侯林义,等. 氯化钆对脓毒症大鼠急性肺损伤的肺保护作用[J].中国医药指南, 2013,11(16):403-404.
[10]Simpson R, Alon R, Kobzik L, et al. Neutrophil and nonneutrophil-mediated injury in intestinal ischemia- reperfusion. Ann Surg. 1993;218(4):444-253.
[11]Xiao F, Eppihimer MJ, Young JA, et al. Lung neutrophil retention and injury after intestinal ischemia/reperfusion. Microcirculation. 1997;4(3):359-367.
[12]孙中吉,卢青,李银平.急性呼吸窘迫综合征发病中的细胞因子和炎性介质[J].中华危重病急救医学, 2003,15(3):186-189.
[13]Kooguchi K, Hashimoto S, Kobayashi A, et al. Role of alveolar macrophages in initiation and regulation of inflammation in Pseudomonas aeruginosa pneumonia. Infect Immun. 1998; 66(7):3164-3169.
[14]Maus UA, Koay MA, Delbeck T, et al. Role of resident alveolar macrophages in leukocyte traffic into the alveolar air space of intact mice. Am J Physiol Lung Cell Mol Physiol. 2002;282(6): L1245-1252.
[15]郭振辉,洪新,毛宝龄,等.核因子-κΒ活化在脓毒症急性肺损伤发病中的作用[J].中国危重病急救医学,2000,12(6):334-337.
[16]Hu X, Qian S, Xu F, et al. Incidence, management and mortality of acute hypoxemic respiratory failure and acute respiratory distress syndrome from a prospective study of Chinese paediatric intensive care network. Acta Paediatr. 2010;99(5):715-721.
[17]董亮,邱海波.急性肺损伤的治疗进展[J].中华急诊医学杂志, 2012,21(3):235-238.
[18]Gupta N, Su X, Popov B, et al. Intrapulmonary delivery of bone marrow-derived mesenchymal stem cells improves survival and attenuates endotoxin-induced acute lung injury in mice. J Immunol. 2007;179(3):1855-1863.
[19]黄杨,梁继河,贾斌,等.白介素10和一氧化氮对内毒素诱导肺泡巨噬细胞核因子κB活化及肿瘤坏死因子释放的调节[J].中国危重病急救医学,2001,13(5):284-286.
[20]Németh K, Leelahavanichkul A, Yuen PS, et al. Bone marrow stromal cells attenuate sepsis via prostaglandin E(2)-dependent reprogramming of host macrophages to increase their interleukin-10 production. Nat Med. 2009;15(1):42-49.
[21]Tai WL, Dong ZX, Zhang DD, et al. Therapeutic effect of intravenous bone marrow-derived mesenchymal stem cell transplantation on early-stage LPS-induced acute lung injury in mice. Nan Fang Yi Ke Da Xue Xue Bao. 2012;32(3):283-290.
[22]Rojas M, Xu J, Woods CR, et al. Bone marrow-derived mesenchymal stem cells in repair of the injured lung. Am J Respir Cell Mol Biol. 2005;33(2):145-152.