Immunophenotypical Characterization of M1/M2 Macrophages and Lymphocytes in Cisplatin-Induced Rat Progressive Renal Fibrosis

Abstract

Renal fibrosis is regarded as the common final pathway leading to chronic kidney diseases; macrophages and myofibroblasts play important roles in the development of fibrosis. F344 rats were injected once with cisplatin (CDDP; 6 mg/kg BW) for renal lesions. Here, immunophenotypical characteristics of macrophages and lymphocytes in CDDP-induced rat renal lesions were investigated histopathologically; the CDDP-induced renal lesions consisted of tissue damage at the early-stage, worsen the damage and commencement of interstitial fibrosis at the mid-stage, and progressive fibrosis at the late stage; the KIM-1 expression and α-SMA+ myofibroblast area reflected renal tubular damage/abnormal regeneration and renal interstitial fibrosis, respectively. CD68+ M1 macrophages began to increase at the mid-stage, with increased mRNA expressions of M1-related cytokines (INF-γ, TNF-α and IL-6), and then slightly decreased at the late-stage. CD163+ M2 macrophages showed a gradually increased number at the mid- and late-stages, accompanied by increased TGF-β1 mRNA expression (a fibrogenic factor). Double immunofluorescence using fibrotic samples at the late-stage revealed that 62.0–78.0% of CD68+ M1 macrophages co-expressed CD163, indicating that M1/M2 macrophages may contribute to progressive renal fibrosis in cooperation; further, MHC class II-expressing macrophages had a tendency towards M1 polarization, whereas CD204-expressing macrophages towards M2 polarization. In addition, CD4+ and CD8+ T cells were increased at the late-stage. Collectively, progressive renal interstitial fibrosis may be developed by complicated mechanisms that arose via interaction of M1/M2 macrophages (inflammatory for M1 and anti-inflammatory for M2) and T cells reacting to CD4 (for helper) and CD8 (for cytotoxicity). This study would provide some information on the pathogenesis of renal fibrosis based on inflammatory cells.

Role of Macrophages and Related Cytokines in Kidney Disease

“…Early studies described that CD163 expression is a hallmark of the wound healing macrophage, related to the resolution phase of inflammation after cardiopulmonary bypass surgery ( 122 ) and in human inflammatory skin disease induced by Cantharidin ( 123 ). Some studies in animal models of renal damage pointed out the key role of CD163 in the beginning and progression of renal disease ( 124 – 126 ). In AKI induced by experimental rhabdomyolysis increased levels of M1 macrophages were observed in the early pathological stages.…”

“…Moreover, peritoneal macrophages stimulated by myoglobin can induce fibrosis through the regulation of profibrotic mediators, such as TGF-β and CTGF/CCN2 ( 124 ). In cisplatin-induced nephrotoxic damage in rats increased levels of CD163 M2 macrophages associated to fibrosis were found ( 126 ). Other study in Lupus nephritis (LN) in mice described an increased CD163 + /CD68 + cell ratio.…”

Local Inflammatory Response after Intramuscularly Implantation of Anti-Adhesive Plasma-Fluorocarbon-Polymer Coated Ti6AI4V Discs in Rats

“…It should also be taken into consideration that in the present study CD68-positive stained cells were designated as pro-inflammatory M1-like cells and CD163-positive cells were designated as anti-inflammatory M2 macrophages. Until the present day, it is not possible to clearly differentiate histologically between M1 and M2 macrophages in rat tissue using only two markers, since there is no clear marker that is exclusively expressed on only one of the macrophage phenotypes [ 33 , 34 , 35 ].…”

Immunophenotypic analysis of the distribution of hepatic macrophages, lymphocytes and hepatic stellate cells in the adult rat liver

“…The hepatic stellate cells have participated in hepatic lesions induced by D‐galactosamine (Rahman et al., 2018) and thioacetamide (Tennakoon et al.,2015). Immunohistochemical staining has been widely used to identify cell phenotypes, cell receptors, cytokines and chemokines (Hall & Rojko, 1996; Nakagawa et al., 2021). Therefore, to establish baseline data and clarify the cell‐specific distribution of hepatic macrophages, lymphocytes, hepatic stellate cells and cholangiocytes in liver parenchyma and Glisson's sheath of rats will be investigated immunohistochemical analysis.…”