Iguratimod Attenuates Macrophage Polarization and Antibody-Mediated Rejection After Renal Transplant by Regulating KLF4

Zhou Hang; Jintao Wei; Ming Zheng; Zeping Gui; Hao Chen; Li Sun; Shuang Fei; Zhijian Han; Jun Tao; Zijie Wang; Ruoyun Tan; Min Gu

doi:10.3389/fphar.2022.865363

Iguratimod Attenuates Macrophage Polarization and Antibody-Mediated Rejection After Renal Transplant by Regulating KLF4

Front Pharmacol. 2022 May 9:13:865363. doi: 10.3389/fphar.2022.865363. eCollection 2022.

Authors

Zhou Hang¹, Jintao Wei², Ming Zheng³, Zeping Gui¹, Hao Chen³, Li Sun³, Shuang Fei³, Zhijian Han³, Jun Tao³, Zijie Wang³, Ruoyun Tan³, Min Gu^{1

3}

Affiliations

¹ Department of Urology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China.
² Department of Emergency Medicine, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
³ Department of Urology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

Abstract

Background: This study aimed to explore the effect and mechanism of iguratimod (IGT) on M1 macrophage polarization and antibody-mediated rejection (ABMR) after renal transplant. Methods: Bioinformatics analysis was performed using three public databases derived from the GEO database. Sprague-Dawley (SD) rats were pre-sensitized with donors of Wistar rats in skin transplantation and a rat renal transplant ABMR model was established from the donors to skin pre-sensitized recipients. Subsequently, IGT was treated on the ABMR model. Routine staining and immunofluorescence (IF) staining were performed to observe the pathological changes in each group and flow cytometry was performed to detect the changes of DSA titers in peripheral blood. In addition, bone-marrow-derived macrophage (BMDM) was extracted and interfered with IGT to explore the effect of IGT in vivo. PCR, IF staining, and Western blot were used to detect the expression of related genes and proteins. Results: Bioinformatics analysis revealed that several immune cells were significantly infiltrated in the ABMR allograft, while M1 macrophage was noticed with the most significance. Results of IF staining and PCR proved the findings of the bioinformatics analysis. Based on this, IGT was observed to significantly attenuate the degree of peritubular capillary vasculitis and arteriolitis in the rat renal transplant ABMR model, whereas it decreases the expression of C4d and reduces the titer of DSA. Results in vitro suggested that M1 macrophage-related transcripts and proteins were significantly reduced by the treatment of IGT in a dose- and time-dependent manner. Furthermore, IGT intervention could remarkably decrease the expression of KLF4. Conclusion: Polarization of M1 macrophages may aggravate ABMR after renal transplant by promoting DSA-mediated endothelial cell injury, and IGT may attenuate the pathogenesis of ABMR by targeting KLF4.

Keywords: KLF4; antibody-mediated rejection; iguratimod; kidney transplant; macrophage polarization.