Aging results in accumulation of M1 and M2 hepatic macrophages and a differential response to gadolinium chloride

  • Steven A. BloomerEmail author
  • Eric D. Moyer
  • Kyle E. Brown
  • Kevin C. Kregel
Original Paper


Macrophages have vital roles in innate immunity by modulating the inflammatory response via their ability to alter their phenotype from pro-inflammatory (M1) to anti-inflammatory (M2). Aging increases activation of the innate immune system, and macrophage numbers increase in the aged liver. Since macrophages also produce free radical molecules, they are a potential source of age-related oxidative injury in the liver. This study evaluated macrophage phenotype in the aged liver and whether the increase in the number of macrophages with aging is associated with enhanced hepatic oxidative stress. Hepatic macrophage phenotype and oxidative stress were evaluated 2 days after a single intraperitoneal injection of saline or gadolinium chloride (GdCl3, 10 mg/kg) in young (6 months) and aged (24 months) Fischer 344 rats. GdCl3 has been shown to decrease the expression of macrophage-specific markers and impair macrophage phagocytosis in the liver. Saline-treated aged rats demonstrated greater numbers of both M1 (HO-1+/iNOS+) and M2 (HO-1+/CD163+) macrophages, without evidence of a phenotypic shift. GdCl3 did not alter levels of dihydroethidium fluorescence or malondialdehyde, suggesting that macrophages are not a major contributor to steady-state levels of oxidative stress. However, GdCl3 decreased M1 and M2 macrophage markers in both age groups, an effect that was attenuated in aged rats. In old animals, GdCl3 decreased iNOS expression to a greater extent than HO-1 or CD163. These results suggest a novel effect of aging on macrophage biology and that GdCl3 shifts hepatic macrophage polarization to the M2 phenotype in aged animals.


Fluorescence microscopy Immunolabelling iNOS Kupffer cells Oxidative stress Inflammation 



We thank Tom Moninger at the University of Iowa Central Microscopy Research Facility for assistance with confocal microscopy.


SAB was supported by a Faculty Development Grant from Penn State Abington. EDM was supported by research funds from the Schreyer Honors College of the Pennsylvania State University, and an Erickson Summer Discovery Grant from the Office of Undergraduate Research at Penn State. This work was also supported by NIH grant AG12350 to KCK. No funding source had a role in study design, data collection, analysis or interpretation of the data, in the writing of the final report, or the decision to submit the article for publication.

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no competing interests.

Ethical approval

All applicable national and institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the University of Iowa (University of Iowa Institutional Animal Care and Use Committee, ACURF #0606117).

Supplementary material

418_2019_1827_MOESM1_ESM.tif (2.6 mb)
Supplementary material 1 (TIFF 2631 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Science and EngineeringPenn State University, Abington CollegeAbingtonUSA
  2. 2.Iowa City Veterans Administration Medical CenterIowa CityUSA
  3. 3.Division of Gastroenterology-HepatologyUniversity of Iowa Roy J. and Lucille A. Carver College of MedicineIowa CityUSA
  4. 4.Program in Free Radical and Radiation BiologyUniversity of Iowa Roy J. and Lucille A. Carver College of MedicineIowa CityUSA
  5. 5.Department of Health and Human PhysiologyThe University of IowaIowa CityUSA

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