自体来源诱导性多能干细胞移植治疗大鼠急性肺损伤
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摘要
背景:干细胞治疗急性肺损伤的研究主要集中在胚胎干细胞和间充质干细胞,有关自体来源诱导性多能干细胞的研究鲜有报道。目的:探讨自体皮肤成纤维细胞来源诱导性多能干细胞尾静脉移植治疗大鼠急性肺损伤的可能性。方法:将24只提取皮肤组织制备诱导性多能干细胞的SD大鼠(北京维通利华实验动物有限公司提供)随机分为3组,对照组腹腔注射生理盐水,模型组和实验组腹腔注射脂多糖制备急性肺损伤模型,造模24 h后,对照组和模型组尾静脉注射PBS,实验组尾静脉注射诱导性多能干细胞悬液。治疗7 d后,观察肺组织形态学、肺组织湿/干质量比、病理损伤程度评分、血清白细胞介素1β、白细胞介素6和肿瘤坏死因子α水平变化。结果与结论:(1)治疗7d后,实验组肺组织损伤明显改善、肺间质水肿减轻、肺泡间隔变薄、毛细血管充血减轻、炎性细胞浸润减少、部分肺泡内的渗出物减少;(2)治疗7 d后,模型组和实验组肺组织湿/干质量比、病理损伤评分明显高于对照组(P <0.01),实验组肺组织湿/干质量比、病理损伤评分明显低于模型组,差异有显著性意义(P <0.01);(3)治疗7 d后,模型组和实验组血清白细胞介素1β、白细胞介素6和肿瘤坏死因子α水平明显高于对照组(P<0.01),实验组血清白细胞介素1β、白细胞介素6和肿瘤坏死因子α水平明显低于模型组,差异均有显著性意义(P <0.01);(4)实验结果表明,自体皮肤成纤维细胞来源的诱导性多能干细胞移植可有效减轻脂多糖诱导的大鼠急性肺损伤,降低血清炎性因子水平。
BACKGROUND: Embryonic stem cells and mesenchymal stem cells are two main cell sources for stem cell transplantation in the treatment of acute lung injury. There are few reports on the study of autologous induced pluripotent stem cells in the treatment of acute lung injury. OBJECTIVE: To investigate the possibility of induced pluripotent stem cells derived from autologous dermal fibroblasts injected through the caudal vein in the treatment of acute lung injury in rats. METHODS: Twenty-four Sprague-Dawley rats(provided by Beijing Vital River Laboratory Animal Technology Co., Ltd.) were randomly divided into three groups. The control group was intraperitoneally injected with normal saline, and the model group and the experimental group were intraperitoneally injected with lipopolysaccharide to make acute lung injury models in rats. At 24 hours after modeling, phosphate buffer solution was injected into the tail vein of the rats in the control and model groups, while the rats in the experimental group were given induced pluripotent stem cell suspension by the tail vein. The changes of lung tissue morphology, lung wet/dry weight ratio, pathological injury score, serum interleukin 1beta, interleukin 6 and tumor necrosis factor alpha levels were observed at 7 days after treatment. RESULTS AND CONCLUSION:(1) At 7 days after treatment, pulmonary interstitial edema, alveolar septum thickening, inflammatory cell infiltration, capillary congestion, irregular alveolar morphology, and exudate in the alveolar cavity were significantly improved in the experimental group.(2) At 7 days after treatment, the wet/dry weight ratio of lung tissue in the model and experimental groups was significantly higher than that in the control group, but the wet/dry weight ratio of lung tissue and pathological injury score in the experimental group were significantly lower than those in the model group(P < 0.01).(3) At 7 days after treatment, the levels of serum interleukin 1beta, interleukin 6 and tumor necrosis factor alpha were ranked as follows: model group > experimental group > control group, and there were significant differences between groups(P < 0.01). To conclude, the transplantation of induced pluripotent stem cells derived from autologous dermal fibroblasts can effectively alleviate acute lung injury and reduce serum inflammatory factor levels in rats.
BACKGROUND: Embryonic stem cells and mesenchymal stem cells are two main cell sources for stem cell transplantation in the treatment of acute lung injury. There are few reports on the study of autologous induced pluripotent stem cells in the treatment of acute lung injury. OBJECTIVE: To investigate the possibility of induced pluripotent stem cells derived from autologous dermal fibroblasts injected through the caudal vein in the treatment of acute lung injury in rats. METHODS: Twenty-four Sprague-Dawley rats(provided by Beijing Vital River Laboratory Animal Technology Co., Ltd.) were randomly divided into three groups. The control group was intraperitoneally injected with normal saline, and the model group and the experimental group were intraperitoneally injected with lipopolysaccharide to make acute lung injury models in rats. At 24 hours after modeling, phosphate buffer solution was injected into the tail vein of the rats in the control and model groups, while the rats in the experimental group were given induced pluripotent stem cell suspension by the tail vein. The changes of lung tissue morphology, lung wet/dry weight ratio, pathological injury score, serum interleukin 1beta, interleukin 6 and tumor necrosis factor alpha levels were observed at 7 days after treatment. RESULTS AND CONCLUSION:(1) At 7 days after treatment, pulmonary interstitial edema, alveolar septum thickening, inflammatory cell infiltration, capillary congestion, irregular alveolar morphology, and exudate in the alveolar cavity were significantly improved in the experimental group.(2) At 7 days after treatment, the wet/dry weight ratio of lung tissue in the model and experimental groups was significantly higher than that in the control group, but the wet/dry weight ratio of lung tissue and pathological injury score in the experimental group were significantly lower than those in the model group(P < 0.01).(3) At 7 days after treatment, the levels of serum interleukin 1beta, interleukin 6 and tumor necrosis factor alpha were ranked as follows: model group > experimental group > control group, and there were significant differences between groups(P < 0.01). To conclude, the transplantation of induced pluripotent stem cells derived from autologous dermal fibroblasts can effectively alleviate acute lung injury and reduce serum inflammatory factor levels in rats.
引文
[1]刘玉静,曹永梅,平凤,等.LPS诱导大鼠急性肺损伤后早期肺纤维化进展过程中肺组织AT1R/Mas受体表达的动态变化[J].上海交通大学学报(医学版),2016,36(5):636-641.
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[6]朱峰,胡贞贞,郭光华.肺外成体干细胞移植治疗急性肺损伤研究进展[J].第二军医大学学报,2009,30(6):713-716.
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[11]Xiang B,Chen L,Wang X,et al.Transplantation of Menstrual Blood-Derived Mesenchymal Stem Cells Promotes the Repair of LPS-Induced Acute Lung Injury.Int J Mol Sci.2017;18(4):E689.
[12]Su VY,Chiou SH,Lin CS,et al.Induced pluripotent stem cells reduce neutrophil chemotaxis via activating GRK2 in endotoxin-induced acute lung injury.Respirology.2017;22(6):1156-1164.
[13]Yang KY,Shih HC,How CK,et al.IV delivery of induced pluripotent stem cells attenuates endotoxin-induced acute lung injury in mice.Chest.2011;140(5):1243-1253.
[14]刘通,张铮.诱导性多能干细胞与心血管疾病[J].心血管病学进展,2011,32(6):873-876.
[15]王建军,赵平,靳雪源,等.诱导性多能干细胞治疗小鼠肝损伤的初步研究[J].中国肝脏病杂志,2016,8(2):39-43.
[16]朱世琪,李峰.诱导性多能干细胞在阿尔茨海默病治疗研究中的应用进展[J].中国组织工程研究,2017,21(21):3426-3431.
[17]任佩佩,樊晋宇,丰慧根,等.诱导性多能干细胞技术在帕金森病研究中的进展[J].医学研究生学报,2016,29(7):770-774.
[18]李萌萌,周建业,宋江平.诱导性多能干细胞在心肌病中的应用[J].中国循环杂志,2017,32(9):934-936.
[19]王欢,方煌,李潇,等.人诱导性多能干细胞的无饲养层培养及鉴定[J].骨科,2016,7(1):49-53.
[20]Yao L,Chen R,Wang P,et al.Generation of induced pluripotent stem cells with high efficiency from human embryonic renal cortical cells.Am J Transl Res.2016;8(11):4982-4993.
[21]曲泽澎,管圆,崔璐,等.大鼠诱导性多能干细胞向神经前体细胞诱导分化研究[J].同济大学学报(医学版),2013,34(3):1-6,11.
[22]陈旭东,范文娟,袁科理,等.不同培养条件对小鼠诱导性多能干细胞分化为神经元样细胞的影响[J].细胞与分子免疫学杂志,2015,31(9):1216-1223.
[23]高鹏飞,张锋,程彬彬,等.核因子-κB介导脂多糖诱导急性肺损伤大鼠高迁移率族蛋白B1的表达[J].临床麻醉学杂志,2015,31(3):278-281.
[24]Wang D,Morales JE,Calame DG,et al.Transplantation of human embryonic stem cell-derived alveolar epithelial type IIcells abrogates acute lung injury in mice.Mol Ther.2010;18(3):625-634.
[25]夏彦民,王文辰,张鸿,等.人羊膜上皮细胞气管内滴注对脂多糖诱导小鼠急性肺损伤的保护作用[J].细胞与分子免疫学杂志,2017,33(1):7-11.
[26]崔蕾,雷鑫,牛玉虎,等.人脐带间充质干细胞移植治疗大鼠急性肺损伤[J].中国组织工程研究,2013,17(14):2563-2569.
[27]Mikawa K,Nishina K,Takao Y,et al.ONO-1714,a nitric oxide synthase inhibitor,attenuates endotoxin-induced acute lung injury in rabbits.Anesth Analg.2003;97(6):1751-1755.
[28]Xin X,Yan L,Guangfa Z,et al.Mesenchymal Stem Cells Promoted Lung Wound Repair through Hox A9 during Endotoxemia-Induced Acute Lung Injury.Stem Cells Int.2017;2017:3648020.
[29]张自丽,刘慧莹,柏长青.间充质干细胞治疗急性肺损伤的研究进展[J].临床肺科杂志,2015,20(11):2096-2097.
[30]王婷,梁华平,柴鉴深,等.三种大鼠急性肺损伤模型的比较[J].成都医学院学报,2016,11(1):5-9.
[31]温保强,刘江华.LPS诱导鼠急性肺损伤模型的评价分析[J].中国医学创新,2017,14(26):137-140.
[32]佟昌慈,柳云恩,张玉彪,等.脂肪干细胞治疗急性肺损伤研究进展[J].创伤与急危重病医学,2017,5(5):292-297.
[33]Pedrazza L,Cunha AA,Luft C,et al.Mesenchymal stem cells improves survival in LPS-induced acute lung injury acting through inhibition of NETs formation.J Cell Physiol.2017;232(12):3552-3564.
[34]Li D,Pan X,Zhao J,et al.Bone Marrow Mesenchymal Stem Cells Suppress Acute Lung Injury Induced by Lipopolysaccharide Through Inhibiting the TLR2,4/NF-κBPathway in Rats with Multiple Trauma.Shock.2016;45(6):641-646.
[35]Hao Q,Zhu YG,Monsel A,et al.Study of Bone Marrow and Embryonic Stem Cell-Derived Human Mesenchymal Stem Cells for Treatment of Escherichia coli Endotoxin-Induced Acute Lung Injury in Mice.Stem Cells Transl Med.2015;4(7):832-840.
[36]Zhang S,Danchuk SD,Bonvillain RW,et al.Interleukin 6mediates the therapeutic effects of adipose-derived stromal/stem cells in lipopolysaccharide-induced acute lung injury.Stem Cells.2014;32(6):1616-1628.
[37]Li J,Huang S,Wu Y,et al.Paracrine factors from mesenchymal stem cells:a proposed therapeutic tool for acute lung injury and acute respiratory distress syndrome.Int Wound J.2014;11(2):114-121.
[38]Xu F,Hu Y,Zhou J,et al.Mesenchymal stem cells in acute lung injury:are they ready for translational medicine.J Cell Mol Med.2013;17(8):927-935.
[39]Martínez-González I,Roca O,Masclans JR,et al.Human mesenchymal stem cells overexpressing the IL-33 antagonist soluble IL-1 receptor-like-1 attenuate endotoxin-induced acute lung injury.Am J Respir Cell Mol Biol.2013;49(4):552-562.
[40]Li JW,Wu X.Mesenchymal stem cells ameliorate LPS-induced acute lung injury through KGF promoting alveolar fluid clearance of alveolar type II cells.Eur Rev Med Pharmacol Sci.2015;19(13):2368-2378.
[2]鲁刚,马奎,付小兵.脐带间充质干细胞对创伤后早期急性肺损伤作用的研究[J].感染、炎症、修复,2013,14(1):3-6.
[3]Matthay MA,Thompson BT,Read EJ,et al.Therapeutic potential of mesenchymal stem cells for severe acute lung injury.Chest.2010;138(4):965-972.
[4]Cribbs SK,Martin GS.Stem cells in sepsis and acute lung injury.Am J Med Sci.2011;341(4):325-332.
[5]Lee JW,Gupta N,Serikov V,et al.Potential application of mesenchymal stem cells in acute lung injury.Expert Opin Biol Ther.2009;9(10):1259-1270.
[6]朱峰,胡贞贞,郭光华.肺外成体干细胞移植治疗急性肺损伤研究进展[J].第二军医大学学报,2009,30(6):713-716.
[7]Soh BS,Zheng D,Li Yeo JS,et al.CD166(pos)subpopulation from differentiated human ES and iPS cells support repair of acute lung injury.Mol Ther.2012;20(12):2335-2346.
[8]Zhang Z,Li W,Heng Z,et al.Combination therapy of human umbilical cord mesenchymal stem cells and FTY720attenuates acute lung injury induced by lipopolysaccharide in a murine model.Oncotarget.2017;8(44):77407-77414.
[9]Zhu H,Xiong Y,Xia Y,et al.Therapeutic Effects of Human Umbilical Cord-Derived Mesenchymal Stem Cells in Acute Lung Injury Mice.Sci Rep.2017;7:39889.
[10]Taki T,Masumoto H,Funamoto M,et al.Fetal mesenchymal stem cells ameliorate acute lung injury in a rat cardiopulmonary bypass model.J Thorac Cardiovasc Surg.2017;153(3):726-734.
[11]Xiang B,Chen L,Wang X,et al.Transplantation of Menstrual Blood-Derived Mesenchymal Stem Cells Promotes the Repair of LPS-Induced Acute Lung Injury.Int J Mol Sci.2017;18(4):E689.
[12]Su VY,Chiou SH,Lin CS,et al.Induced pluripotent stem cells reduce neutrophil chemotaxis via activating GRK2 in endotoxin-induced acute lung injury.Respirology.2017;22(6):1156-1164.
[13]Yang KY,Shih HC,How CK,et al.IV delivery of induced pluripotent stem cells attenuates endotoxin-induced acute lung injury in mice.Chest.2011;140(5):1243-1253.
[14]刘通,张铮.诱导性多能干细胞与心血管疾病[J].心血管病学进展,2011,32(6):873-876.
[15]王建军,赵平,靳雪源,等.诱导性多能干细胞治疗小鼠肝损伤的初步研究[J].中国肝脏病杂志,2016,8(2):39-43.
[16]朱世琪,李峰.诱导性多能干细胞在阿尔茨海默病治疗研究中的应用进展[J].中国组织工程研究,2017,21(21):3426-3431.
[17]任佩佩,樊晋宇,丰慧根,等.诱导性多能干细胞技术在帕金森病研究中的进展[J].医学研究生学报,2016,29(7):770-774.
[18]李萌萌,周建业,宋江平.诱导性多能干细胞在心肌病中的应用[J].中国循环杂志,2017,32(9):934-936.
[19]王欢,方煌,李潇,等.人诱导性多能干细胞的无饲养层培养及鉴定[J].骨科,2016,7(1):49-53.
[20]Yao L,Chen R,Wang P,et al.Generation of induced pluripotent stem cells with high efficiency from human embryonic renal cortical cells.Am J Transl Res.2016;8(11):4982-4993.
[21]曲泽澎,管圆,崔璐,等.大鼠诱导性多能干细胞向神经前体细胞诱导分化研究[J].同济大学学报(医学版),2013,34(3):1-6,11.
[22]陈旭东,范文娟,袁科理,等.不同培养条件对小鼠诱导性多能干细胞分化为神经元样细胞的影响[J].细胞与分子免疫学杂志,2015,31(9):1216-1223.
[23]高鹏飞,张锋,程彬彬,等.核因子-κB介导脂多糖诱导急性肺损伤大鼠高迁移率族蛋白B1的表达[J].临床麻醉学杂志,2015,31(3):278-281.
[24]Wang D,Morales JE,Calame DG,et al.Transplantation of human embryonic stem cell-derived alveolar epithelial type IIcells abrogates acute lung injury in mice.Mol Ther.2010;18(3):625-634.
[25]夏彦民,王文辰,张鸿,等.人羊膜上皮细胞气管内滴注对脂多糖诱导小鼠急性肺损伤的保护作用[J].细胞与分子免疫学杂志,2017,33(1):7-11.
[26]崔蕾,雷鑫,牛玉虎,等.人脐带间充质干细胞移植治疗大鼠急性肺损伤[J].中国组织工程研究,2013,17(14):2563-2569.
[27]Mikawa K,Nishina K,Takao Y,et al.ONO-1714,a nitric oxide synthase inhibitor,attenuates endotoxin-induced acute lung injury in rabbits.Anesth Analg.2003;97(6):1751-1755.
[28]Xin X,Yan L,Guangfa Z,et al.Mesenchymal Stem Cells Promoted Lung Wound Repair through Hox A9 during Endotoxemia-Induced Acute Lung Injury.Stem Cells Int.2017;2017:3648020.
[29]张自丽,刘慧莹,柏长青.间充质干细胞治疗急性肺损伤的研究进展[J].临床肺科杂志,2015,20(11):2096-2097.
[30]王婷,梁华平,柴鉴深,等.三种大鼠急性肺损伤模型的比较[J].成都医学院学报,2016,11(1):5-9.
[31]温保强,刘江华.LPS诱导鼠急性肺损伤模型的评价分析[J].中国医学创新,2017,14(26):137-140.
[32]佟昌慈,柳云恩,张玉彪,等.脂肪干细胞治疗急性肺损伤研究进展[J].创伤与急危重病医学,2017,5(5):292-297.
[33]Pedrazza L,Cunha AA,Luft C,et al.Mesenchymal stem cells improves survival in LPS-induced acute lung injury acting through inhibition of NETs formation.J Cell Physiol.2017;232(12):3552-3564.
[34]Li D,Pan X,Zhao J,et al.Bone Marrow Mesenchymal Stem Cells Suppress Acute Lung Injury Induced by Lipopolysaccharide Through Inhibiting the TLR2,4/NF-κBPathway in Rats with Multiple Trauma.Shock.2016;45(6):641-646.
[35]Hao Q,Zhu YG,Monsel A,et al.Study of Bone Marrow and Embryonic Stem Cell-Derived Human Mesenchymal Stem Cells for Treatment of Escherichia coli Endotoxin-Induced Acute Lung Injury in Mice.Stem Cells Transl Med.2015;4(7):832-840.
[36]Zhang S,Danchuk SD,Bonvillain RW,et al.Interleukin 6mediates the therapeutic effects of adipose-derived stromal/stem cells in lipopolysaccharide-induced acute lung injury.Stem Cells.2014;32(6):1616-1628.
[37]Li J,Huang S,Wu Y,et al.Paracrine factors from mesenchymal stem cells:a proposed therapeutic tool for acute lung injury and acute respiratory distress syndrome.Int Wound J.2014;11(2):114-121.
[38]Xu F,Hu Y,Zhou J,et al.Mesenchymal stem cells in acute lung injury:are they ready for translational medicine.J Cell Mol Med.2013;17(8):927-935.
[39]Martínez-González I,Roca O,Masclans JR,et al.Human mesenchymal stem cells overexpressing the IL-33 antagonist soluble IL-1 receptor-like-1 attenuate endotoxin-induced acute lung injury.Am J Respir Cell Mol Biol.2013;49(4):552-562.
[40]Li JW,Wu X.Mesenchymal stem cells ameliorate LPS-induced acute lung injury through KGF promoting alveolar fluid clearance of alveolar type II cells.Eur Rev Med Pharmacol Sci.2015;19(13):2368-2378.