Effect of polarized bone marrow-derived macrophage transplantation on the progression of CCl<sub>4</sub>-induced liver fibrosis in rats

Xun JIAN; Danyang WANG; Yannan XU; Jiamei CHEN; Wei LIU; Gaofeng CHEN; Hua ZHANG; Ping LIU; Yongping MU

doi:10.3969/j.issn.1001-5256.2021.12.020

Volume 37 Issue 12

Dec. 2021

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Article Navigation > Journal of Clinical Hepatology > 2021 > 37(12): 2830-2837

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Effect of polarized bone marrow-derived macrophage transplantation on the progression of CCl₄-induced liver fibrosis in rats

DOI: 10.3969/j.issn.1001-5256.2021.12.020

Key Laboratory of Liver and Kidney Diseases (Ministry of Education), Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shanghai 201203, China

Research funding:

The National Natural Science Foundation of China (81573948);

The National Natural Science Foundation of China (81874390)

Received Date: 2021-04-22
Accepted Date: 2021-07-28
Published Date: 2021-12-20

Abstract

Abstract

Objective To investigate the effect of polarized bone marrow-derived macrophage (BMDM) transplantation on the progression of CCl₄-induced liver fibrosis in rats. Methods Rat BMDMs were isolated and induced to differentiate into M1 phenotype (M1-BMDM) by lipopolysaccharide (5 ng/mL) or M2 phenotype (M2-BMDM) by the supernatant of L929 cells. A rat model of liver fibrosis was established by subcutaneous injection of 30% CCl₄ for 6 weeks, and at week 7, the model rats were randomly divided into model control group (M group), M1-BMDM group, and M2-BMDM group and were given a single injection of normal saline, M1-BMDM, and M2-BMDM, respectively, via the caudal vein, and subcutaneous injection of 30% CCl₄ was given until the end of week 9. Related indices were observed, including liver function, liver histopathology, hydroxyproline (Hyp) content in liver tissue, hepatic stellate cell activation, liver fibrosis, and expression of inflammatory cytokines. The continuous data were expressed as mean±standard deviation; an analysis of variance was used for comparison between multiple groups, and the SNK-q test was used for further comparison between two groups. Results Compared with the M group, both M1-BMDM and M2-BMDM significantly inhibited liver inflammation and liver fibrosis progression and significantly reduced serum alanine aminotransferase and aspartate aminotransferase activities (P < 0.01) and Hyp content in liver tissue (P < 0.05). M1-BMDM and M2-BMDM significantly inhibited the activation of hepatic stellate cells and significantly reduced the mRNA expression levels of TGF-β, Col1A1, and Col4 (all P < 0.05). Both M1-BMDM and M2-BMDM significantly increased the expression level of CD163 protein in liver tissue (P < 0.01), and the M2-BMDM group had a significantly higher level than the M1-BMDM group (P < 0.05); both M1-BMDM and M2-BMDM significantly reduced the mRNA expression levels of MMP-2 and TIMP-1 in liver tissue (P < 0.05) and significantly increased the mRNA expression level of MMP-13 (P < 0.01); in addition, M2-BMDM significantly reduced the expression level of CD68 protein in liver tissue (P < 0.01). Both M1-BMDM and M2-BMDM significantly increased the mRNA expression levels of IL-6 and IL-10 and the protein expression level of albumin in liver tissue (all P < 0.05), and the above indices in the M2-BMDM group were significantly higher than those in the M1-BMDM group (all P < 0.05). Conclusion Both M1-BMDM and M2-BMDM can effectively inhibit the progression of CCl₄-induced liver fibrosis in rats, possibly by inhibiting the activation of hepatic stellate cells and promoting the activation of anti-inflammatory macrophages. Moreover, M2-BMDM can also inhibit the activation of pro-inflammatory macrophages and thus has a better comprehensive intervention effect than M1-BMDM.
- Liver Cirrhosis,
- Hepatic Stellate Cells,
- Macrophages

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References

[1]	RAMACHANDRAN P, DOBIE R, WILSON-KANAMORI JR, et al. Resolving the fibrotic niche of human liver cirrhosis at single-cell level[J]. Nature, 2019, 575(7783): 512-518. DOI: 10.1038/s41586-019-1631-3.
[2]	KARLMARK KR, WEISKIRCHEN R, ZIMMERMANN HW, et al. Hepatic recruitment of the inflammatory Gr1+ monocyte subset upon liver injury promotes hepatic fibrosis[J]. Hepatology, 2009, 50(1): 261-274. DOI: 10.1002/hep.22950.
[3]	VANNELLA KM, WYNN TA. Mechanisms of organ injury and repair by macrophages[J]. Annu Rev Physiol, 2017, 79: 593-617. DOI: 10.1146/annurev-physiol-022516-034356.
[4]	ORECCHIONI M, GHOSHEH Y, PRAMOD AB, et al. Macrophage polarization: Different gene signatures in M1(LPS+) vs. classically and M2(LPS-) vs. alternatively activated macrophages[J]. Front Immunol, 2019, 10: 1084. DOI: 10.3389/fimmu.2019.01084.
[5]	MURRAY PJ, ALLEN JE, BISWAS SK, et al. Macrophage activation and polarization: Nomenclature and experimental guidelines[J]. Immunity, 2014, 41(1): 14-20. DOI: 10.1016/j.immuni.2014.06.008.
[6]	DAVIS BK. Derivation of macrophages from mouse bone marrow[J]. Methods Mol Biol, 2019, 1960: 41-55. DOI: 10.1007/978-1-4939-9167-9_3.
[7]	WATANABE Y, TSUCHIYA A, SEINO S, et al. Mesenchymal stem cells and induced bone marrow-derived macrophages synergistically improve liver fibrosis in mice[J]. Stem Cells Transl Med, 2019, 8(3): 271-284. DOI: 10.1002/sctm.18-0105.
[8]	PINEDA-TORRA I, GAGE M, de JUAN A, et al. Isolation, culture, and polarization of murine bone marrow-derived and peritoneal macrophages[J]. Methods Mol Biol, 2015, 1339: 101-109. DOI: 10.1007/978-1-4939-2929-0_6.
[9]	MILY A, KALSUM S, LORETI MG, et al. Polarization of M1 and M2 human monocyte-derived cells and analysis with flow cytometry upon mycobacterium tuberculosis infection[J]. J Vis Exp, 2020, (163). DOI: 10.3791/61807.
[10]	JAMALL IS, FINELLI VN, QUE HEE SS. A simple method to determine nanogram levels of 4-hydroxyproline in biological tissues[J]. Anal Biochem, 1981, 112(1): 70-75. DOI: 10.1016/0003-2697(81)90261-x.
[11]	MU YP, OGAWA T, KAWADA N. Reversibility of fibrosis, inflammation, and endoplasmic reticulum stress in the liver of rats fed a methionine-choline-deficient diet[J]. Lab Invest, 2010, 90(2): 245-256. DOI: 10.1038/labinvest.2009.123.
[12]	PRADERE JP, KLUWE J, DE MINICIS S, et al. Hepatic macrophages but not dendritic cells contribute to liver fibrosis by promoting the survival of activated hepatic stellate cells in mice[J]. Hepatology, 2013, 58(4): 1461-1473. DOI: 10.1002/hep.26429.
[13]	HUME DA, IRVINE KM, PRIDANS C. The mononuclear phagocyte system: The relationship between monocytes and macrophages[J]. Trends Immunol, 2019, 40(2): 98-112. DOI: 10.1016/j.it.2018.11.007.
[14]	MA PF, GAO CC, YI J, et al. Cytotherapy with M1-polarized macrophages ameliorates liver fibrosis by modulating immune microenvironment in mice[J]. J Hepatol, 2017, 67(4): 770-779. DOI: 10.1016/j.jhep.2017.05.022.
[15]	FRIEDMAN SL. Hepatic stellate cells: Protean, multifunctional, and enigmatic cells of the liver[J]. Physiol Rev, 2008, 88(1): 125-172. DOI: 10.1152/physrev.00013.2007.
[16]	NOVO E, MARRA F, ZAMARA E, et al. Dose dependent and divergent effects of superoxide anion on cell death, proliferation, and migration of activated human hepatic stellate cells[J]. Gut, 2006, 55(1): 90-97. DOI: 10.1136/gut.2005.069633.
[17]	RAMACHANDRAN P, IREDALE JP. Macrophages: Central regulators of hepatic fibrogenesis and fibrosis resolution[J]. J Hepatol, 2012, 56(6): 1417-1419. DOI: 10.1016/j.jhep.2011.10.026.
[18]	FENG M, DING J, WANG M, et al. Kupffer-derived matrix metalloproteinase-9 contributes to liver fibrosis resolution[J]. Int J Biol Sci, 2018, 14(9): 1033-1040. DOI: 10.7150/ijbs.25589.
[19]	GUO J, LUO Y, YIN F, et al. Overexpression of tumor necrosis factor-like ligand 1 A in myeloid cells aggravates liver fibrosis in mice[J]. J Immunol Res, 2019, 2019: 7657294. DOI: 10.1155/2019/7657294.
[20]	GORDON S, MARTINEZ FO. Alternative activation of macrophages: Mechanism and functions[J]. Immunity, 2010, 32(5): 593-604. DOI: 10.1016/j.immuni.2010.05.007.
[21]	TAO Y, WANG M, CHEN E, et al. Liver Regeneration: Analysis of the main relevant signaling molecules[J]. Mediators Inflamm, 2017, 2017: 4256352. DOI: 10.1155/2017/4256352.
[22]	CRESSMAN DE, GREENBAUM LE, DEANGELIS RA, et al. Liver failure and defective hepatocyte regeneration in interleukin-6-deficient mice[J]. Science, 1996, 274(5291): 1379-1383. DOI: 10.1126/science.274.5291.1379.
[23]	TAUB R. Liver regeneration: From myth to mechanism[J]. Nat Rev Mol Cell Biol, 2004, 5(10): 836-847. DOI: 10.1038/nrm1489.
[24]	KANG LI, MARS WM, MICHALOPOULOS GK. Signals and cells involved in regulating liver regeneration[J]. Cells, 2012, 1(4): 1261-1292. DOI: 10.3390/cells1041261.
[25]	ELCHANINOV AV, FATKHUDINOV TK, VISHNYAKOVA PA, et al. Phenotypical and functional polymorphism of liver resident macrophages[J]. Cells, 2019, 8(9): 1032-1053. DOI: 10.3390/cells8091032.
[26]	CAMPANA L, ESSER H, HUCH M, et al. Liver regeneration and inflammation: From fundamental science to clinical applications[J]. Nat Rev Mol Cell Biol, 2021, 22(9): 608-624. DOI: 10.1038/s41580-021-00373-7.
[27]	BARBAY V, HOUSSARI M, MEKKI M, et al. Role of M2-like macrophage recruitment during angiogenic growth factor therapy[J]. Angiogenesis, 2015, 18(2): 191-200. DOI: 10.1007/s10456-014-9456-z.
[28]	DONG X, LIU J, XU Y, et al. Role of macrophages in experimental liver injury and repair in mice[J]. Exp Ther Med, 2019, 17(5): 3835-3847. DOI: 10.3892/etm.2019.7450.

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Effect of polarized bone marrow-derived macrophage transplantation on the progression of CCl4-induced liver fibrosis in rats

DOI: 10.3969/j.issn.1001-5256.2021.12.020

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Effect of polarized bone marrow-derived macrophage transplantation on the progression of CCl₄-induced liver fibrosis in rats