Email Alerts
RSS
Turn off MathJax
Article Contents
Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  >  > Accepted Manuscript
Bai Xiaozhi,Tao Ke,Liu Yang,et al.Effects and underlying mechanism of exosomes of adipose-derived mesenchymal stem cells on acute lung injury of septic mice[J].Chin J Burns Wounds,2024,40(12):1-10.DOI: 10.3760/cma.j.cn501225-20240927-00355.
Citation: Bai Xiaozhi,Tao Ke,Liu Yang,et al.Effects and underlying mechanism of exosomes of adipose-derived mesenchymal stem cells on acute lung injury of septic mice[J].Chin J Burns Wounds,2024,40(12):1-10.DOI: 10.3760/cma.j.cn501225-20240927-00355.
shu

Effects and underlying mechanism of exosomes of adipose-derived mesenchymal stem cells on acute lung injury of septic mice

doi: 10.3760/cma.j.cn501225-20240927-00355
  • 1.

    Department of Burns and Cutaneous Surgery, Burn Center of PLA, the First Affiliated Hospital of Air Force Medical University, Xi'an710032, China

  • 2.

    Department of Wound Repair, Center for Wound Repair and Regenerative Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou325015, China

Funds:

General Program of National Natural Science Foundation of China 82272269

  • Received Date: 2024-09-27
    Available Online: 2024-12-02
    Abstract
  •   Objective  To explore effects and underlying mechanism of exosomes of adipose-derived mesenchymal stem cells (ADSC) on acute lung injury of septic mice.  Methods  The study was an experimental study. ADSC of passages 4-5 were selected, and exosomes in their supernatant were isolated and extracted by differential ultracentrifugation. Exosomes were then used after identification. Twenty-four adult male BALB/c mice were selected and divided into control group, simple cecal ligation and puncture (CLP) group, and CLP+ADSC-exosome group according to the random number table method (the grouping method was the same as below), with 8 mice in each group, which were treated accordingly. At 24 hours after injury, hematoxylin-eosin staining was used to observe the morphology of lung tissues, the in-situ end-labeling method was used to detect the apoptosis of lung tissue cells, the enzyme-linked immunosorbent assay was used to detect the levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the serum of mice, relevant kits were used to detect the contents of malondialdehyde and superoxide dismutase (SOD) in lung tissues, and immunofluorescence method was used to detect the expressions of CD86 and CD206 in lung tissue cells of mice. Mouse macrophage RAW246.7 was taken and divided into control group, simple LPS group, and LPS+ADSC-exosome group, which were treated accordingly. Twelve hours later, the ATP content, the percentage of mitochondrial ROS positive cells, as well as mitochondrial membrane potential in cells were detected by related detection kits. The real-time fluorescence quantitative reverse-ranscription PCR method was used to detect the mRNA expression levels of M1 polarization marker factor inducible nitric oxide synthase (iNOS), M2 polarization marker factor arginase-1 (Arg1), and inflammatory factors TNF-α and IL-1β in cells. Three samples were used for mRNA expression detection, and four samples were used for the detection of other indicators.  Results  At 24 hours after injury, the lung tissue structure of mice in control group was clear and intact without inflammatory cell infiltration. Compared with that in control group, the lung tissue edema as well as the inflammatory cell infiltration of mice was much more obvious in simple CLP group. However, compared with that in simple CLP group, the lung tissue edema in CLP+ADSC-exosome group mice was significantly alleviated, the inflammatory cell infiltration was significantly reduced, and the cell apoptosis and necrosis were significantly improved. Twenty-four hours after injury, compared with that in control group, the contents of TNF-α and IL-1β in the serum of mice in simple CLP group were significantly increased (with t values of 50.82 and 30.81, respectively, P<0.05); compared with that in simple CLP group, the contents of TNF-α and IL-1β in the serum of mice in CLP+ADSC-exosome group were significantly decreased (with t values of 16.36 and 19.25, respectively, P<0.05). Compared with that in control group, the content of malondialdehyde in the lung tissue of mice in simple CLP group was significantly increased (t=9.89, P<0.05), and the content of SOD was significantly decreased (t=5.01, P<0.05); compared with that in simple CLP group, the content of malondialdehyde in the lung tissue of mice in CLP+ADSC-exosome group was significantly decreased (t=4.38, P<0.05), and the content of SOD was significantly increased (t=2.97, P<0.05). Twenty-four hours after injury, compared with that in control group, the number of CD86 positive cells in the lung tissue of mice in simple CLP group was significantly increased, and the number of CD206 positive cells was significantly decreased; compared with that in simple CLP group, the number of CD86 positive cells in the lung tissue of mice in CLP+ADSC-exosome group was significantly decreased, and the number of CD206 positive cells was significantly increased. After 12 hours of culture, compared with that in control group, the ATP content of RAW246.7 cells in simple LPS group was significantly decreased (t=6.28, P<0.05); compared with that in simple LPS group, the ATP content of RAW246.7 cells in LPS+ADSC-exosome group was significantly increased (t=4.01, P<0.05). After 12 hours of culture, compared with 22%±4% in blank control group, 40%±6% of positive cells of mitochondrial reactive oxygen species in RAW246.7 cells in simple LPS group was significantly increased (t=5.04, P<0.05); Compared with that in LPS group, 30%±5% of positive cells of mitochondrial reactive oxygen species in RAW246.7 cells in LPS+ADSC-exosome group was significantly decreased (t=2.65, P<0.05). After 12 hours of culture, compared with that in control group, the mRNA expressions of TNF-α, IL-1β, and iNOS in RAW246.7 cells in simple LPS group were significantly increased (with t values of16.51, 31.04, and 7.70, respectively, P<0.05), and the decrease in the mRNA expression of Arg1 was not statistically significant (P>0.05); compared with that in simple LPS group, the mRNA expressions of TNF-α, IL-1β, and iNOS in RAW246.7 cells in LPS+ADSC-exosome group were significantly decreased (with t values of 11.38, 22.58, and 5.28, respectively, P<0.05), and the mRNA expression of Arg1 was significantly increased (t=7.66, P<0.05).  Conclusions  Human ADSC-exosomes may play a role in improving lung injury in septic mice by improving LPS-induced mitochondrial dysfunction in RAW246.7 cells, inhibiting macrophage polarization to M1, and reducing the inflammatory response.

     

    • FullText(HTML)
  • loading
    • References(39)
  • [1]
    LongME, MallampalliRK, HorowitzJC. Pathogenesis of pneumonia and acute lung injury[J]. Clin Sci (Lond), 2022,136(10):747-769. DOI: 10.1042/CS20210879.
    [2]
    LocatiM, CurtaleG, DiversityMantovani A., mechanisms, and significance of macrophage plasticity[J]. Annu Rev Pathol, 2020,15:123-147. DOI: 10.1146/annurev-pathmechdis-012418-012718.
    [3]
    OishiY, ManabeI. Macrophages in inflammation, repair and regeneration[J]. Int Immunol, 2018,30(11):511-528. DOI: 10.1093/intimm/dxy054.
    [4]
    SchumackerPT, GillespieMN, NakahiraK, et al. Mitochondria in lung biology and pathology: more than just a powerhouse[J]. Am J Physiol Lung Cell Mol Physiol, 2014,306(11):L962-974. DOI: 10.1152/ajplung.00073.2014.
    [5]
    KellnerM, NoonepalleS, LuQ, et al. ROS signaling in the pathogenesis of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS)[J]. Adv Exp Med Biol, 2017,967:105-137. DOI: 10.1007/978-3-319-63245-2_8.
    [6]
    邱煜程, 周显玉, 刘菲, 等. 间充质干细胞及其外泌体在移植中的应用进展[J].组织工程与重建外科杂志,2023,19(2):184-188. DOI: 10.3969/j.issn.1673-0364.2023.02.016.
    [7]
    YuT, LiuH, GaoM, et al. Dexmedetomidine regulates exosomal miR-29b-3p from macrophages and alleviates septic myocardial injury by promoting autophagy in cardiomyocytes via targeting glycogen synthase kinase3β[J/OL]. Burns Trauma, 2024,12:tkae042[2024-09-27].https://pubmed.ncbi.nlm.nih.gov/39502342/. DOI: 10.1093/burnst/tkae042.
    [8]
    蒲倩, 修光辉, 孙洁, 等. 间充质干细胞外泌体在脓毒症多器官功能障碍中作用的研究进展[J].中华危重病急救医学,2021,33(6):757-760. DOI: 10.3760/cma.j.cn121430-20200908-00620.
    [9]
    HuQ, LyonCJ, FletcherJK, et al. Extracellular vesicle activities regulating macrophage- and tissue-mediated injury and repair responses[J]. Acta Pharm Sin B, 2021,11(6):1493-1512. DOI: 10.1016/j.apsb.2020.12.014.
    [10]
    JingW, WangH, ZhanL, et al. Extracellular vesicles, new players in sepsis and acute respiratory distress syndrome[J]. Front Cell Infect Microbiol, 2022,12:853840. DOI: 10.3389/fcimb.2022.853840.
    [11]
    HommaK, BazhanovN, HashimotoK, et al. Mesenchymal stem cell-derived exosomes for treatment of sepsis[J]. Front Immunol, 2023,14:1136964. DOI: 10.3389/fimmu.2023.1136964.
    [12]
    GongT, LiuYT, FanJ. Exosomal mediators in sepsis and inflammatory organ injury: unraveling the role of exosomes in intercellular crosstalk and organ dysfunction[J]. Mil Med Res, 2024,11(1):24. DOI: 10.1186/s40779-024-00527-6.
    [13]
    BaiX, LiJ, LiL, et al. Extracellular vesicles from adipose tissue-derived stem cells affect Notch-miR148a-3p axis to regulate polarization of macrophages and alleviate sepsis in mice[J]. Front Immunol, 2020,11:1391. DOI: 10.3389/fimmu.2020.01391.
    [14]
    DejagerL, PinheiroI, DejonckheereE, et al. Cecal ligation and puncture: the gold standard model for polymicrobial sepsis?[J]. Trends Microbiol, 2011,19(4):198-208. DOI: 10.1016/j.tim.2011.01.001.
    [15]
    JiaoY, ZhangT, ZhangC, et al. Exosomal miR-30d-5p of neutrophils induces M1 macrophage polarization and primes macrophage pyroptosis in sepsis-related acute lung injury[J]. Crit Care, 2021,25(1):356. DOI: 10.1186/s13054-021-03775-3.
    [16]
    BaiX, HeT, LiuY, et al. Acetylation-dependent regulation of notch signaling in macrophages by SIRT1 affects sepsis development[J]. Front Immunol, 2018,9:762. DOI: 10.3389/fimmu.2018.00762.
    [17]
    蔡维霞, 沈括, 曹涛, 等. 人脂肪间充质干细胞来源外泌体对脓毒症小鼠肺血管内皮细胞损伤的影响及其机制[J].中华烧伤与创面修复杂志,2022,38(3):266-275. DOI: 10.3760/cma.j.cn501120-20211020-00362.
    [18]
    ShenK, WangX, WangY, et al. miR-125b-5p in adipose derived stem cells exosome alleviates pulmonary microvascular endothelial cells ferroptosis via Keap1/Nrf2/GPX4 in sepsis lung injury[J]. Redox Biol, 2023,62:102655. DOI: 10.1016/j.redox.2023.102655.
    [19]
    WuH, WangY, ZhangY, et al. Breaking the vicious loop between inflammation, oxidative stress and coagulation, a novel anti-thrombus insight of nattokinase by inhibiting LPS-induced inflammation and oxidative stress[J]. Redox Biol, 2020,32:101500. DOI: 10.1016/j.redox.2020.101500.
    [20]
    XuH, QiQ, YanX. Myricetin ameliorates sepsis-associated acute lung injury in a murine sepsis model[J]. Naunyn Schmiedebergs Arch Pharmacol, 2021,394(1):165-175. DOI: 10.1007/s00210-020-01880-8.
    [21]
    JinC, ChenJ, GuJ, et al. Gut-lymph-lung pathway mediates sepsis-induced acute lung injury[J]. Chin Med J (Engl), 2020,133(18):2212-2218. DOI: 10.1097/CM9.0000000000000928.
    [22]
    YehyaN, SmithL, ThomasNJ, et al. Definition, incidence, and epidemiology of pediatric acute respiratory distress syndrome: from the Second Pediatric Acute Lung Injury Consensus Conference[J]. Pediatr Crit Care Med, 2023,24(12 Suppl 2):S87-98. DOI: 10.1097/PCC.0000000000003161.
    [23]
    赵松韵, 万志杰, 曹曦元, 等. 靶向DNA损伤应答在小细胞肺癌中的作用研究进展[J].解放军医学杂志,2022,47(8):838-844. DOI: 10.11855/j.issn.0577-7402.2022.08.0838.
    [24]
    李林, 邢福席, 付全有, 等. 脓毒症急性肺损伤治疗的研究进展[J].中华医院感染学杂志,2024,34(1):149-155. DOI: 10.11816/cn.ni.2024-236123.
    [25]
    ZhangW, ChenH, XuZ, et al. Liensinine pretreatment reduces inflammation, oxidative stress, apoptosis, and autophagy to alleviate sepsis acute kidney injury[J]. Int Immunopharmacol, 2023,122:110563. DOI: 10.1016/j.intimp.2023.110563.
    [26]
    Bar-OrD, CarrickMM, MainsCW, et al. Sepsis, oxidative stress, and hypoxia: are there clues to better treatment?[J] Redox Rep, 2015,20(5):193-197. DOI: 10.1179/1351000215Y.0000000005.
    [27]
    JoffreJ, HellmanJ. Oxidative stress and endothelial dysfunction in sepsis and acute inflammation[J]. Antioxid Redox Signal, 2021,35(15):1291-1307. DOI: 10.1089/ars.2021.0027.
    [28]
    WangX, ChenS, LuR, et al. Adipose-derived stem cell-secreted exosomes enhance angiogenesis by promoting macrophage M2 polarization in type 2 diabetic mice with limb ischemia via the JAK/STAT6 pathway[J]. Heliyon, 2022,8(11):e11495. DOI: 10.1016/j.heliyon.2022.e11495.
    [29]
    WestAP, BrodskyIE, RahnerC, et al. TLR signalling augments macrophage bactericidal activity through mitochondrial ROS[J]. Nature, 2011,472(7344):476-480. DOI: 10.1038/nature09973.
    [30]
    WestAP, Khoury-HanoldW, StaronM, et al. Mitochondrial DNA stress primes the antiviral innate immune response[J]. Nature, 2015,520(7548):553-557. DOI: 10.1038/nature14156.
    [31]
    WangZ, WhiteA, WangX, et al. Mitochondrial fission mediated cigarette smoke-induced pulmonary endothelial injury[J]. Am J Respir Cell Mol Biol, 2020,63(5):637-651. DOI: 10.1165/rcmb.2020-0008OC.
    [32]
    VidelaLA, MarimánA, RamosB, et al. Standpoints in mitochondrial dysfunction: underlying mechanisms in search of therapeutic strategies[J]. Mitochondrion, 2022,63:9-22. DOI: 10.1016/j.mito.2021.12.006.
    [33]
    HoffmannRF, ZarrintanS, BrandenburgSM, et al. Prolonged cigarette smoke exposure alters mitochondrial structure and function in airway epithelial cells[J]. Respir Res, 2013,14(1):97. DOI: 10.1186/1465-9921-14-97.
    [34]
    GalleyHF. Oxidative stress and mitochondrial dysfunction in sepsis[J]. Br J Anaesth, 2011, 107(1):57-64. DOI: 10.1093/bja/aer093.
    [35]
    XianH, LiuY, Rundberg NilssonA, et al. Metformin inhibition of mitochondrial ATP and DNA synthesis abrogates NLRP3 inflammasome activation and pulmonary inflammation[J]. Immunity, 2021,54(7):1463-1477.e11. DOI: 10.1016/j.immuni.2021.05.004.
    [36]
    ZhongZ, LiangS, Sanchez-LopezE, et al. New mitochondrial DNA synthesis enables NLRP3 inflammasome activation[J]. Nature, 2018,560(7717):198-203. DOI: 10.1038/s41586-018-0372-z.
    [37]
    XuZ, ShenJ, LinL, et al. Exposure to irregular microplastic shed from baby bottles activates the ROS/NLRP3/Caspase-1 signaling pathway, causing intestinal inflammation[J]. Environ Int, 2023,181:108296. DOI: 10.1016/j.envint.2023.108296.
    [38]
    Shang-GuanK, WangM, HtweN, et al. Lipopolysaccharides trigger two successive bursts of reactive oxygen species at distinct cellular locations[J]. Plant Physiol, 2018,176(3):2543-2556. DOI: 10.1104/pp.17.01637.
    [39]
    CaiS, ZhaoM, ZhouB, et al. Mitochondrial dysfunction in macrophages promotes inflammation and suppresses repair after myocardial infarction[J]. J Clin Invest, 2023,133(4)DOI: 10.1172/JCI159498.
    • Relative Articles
    • Supplements(0)
    • Cited By
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Figures(6)  / Tables(2)

    Get Citation
    PDF
    XML

    Article Metrics

    Article views (27) PDF downloads(2) Cited by()
    Proportional views
    Related

    Copyright © Chinese Journal of Burns京ICP备07035254号-14

    E-mail：shaoshangzazhi@163.com

    Website：https://zhsszz.xml-journal.net/

    Supported by: Beijing Renhe Information Technology Co. Ltd    

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return