Glutathione S-transferase Mu 2-transduced mesenchymal stem cells ameliorated anti-glomerular basement membrane antibody-induced glomerulonephritis by inhibiting oxidation and inflammation

Abstract

IntroductionOxidative stress is implicated in tissue inflammation, and plays an important role in the pathogenesis of immune-mediated nephritis. Using the anti-glomerular basement membrane antibody-induced glomerulonephritis (anti-GBM-GN) mouse model, we found that increased expression of glutathione S-transferase Mu 2 (GSTM2) was related to reduced renal damage caused by anti-GBM antibodies. Furthermore, mesenchymal stem cell (MSC)-based therapy has shed light on the treatment of immune-mediated kidney diseases. The aim of this study was to investigate if MSCs could be utilized as vehicles to deliver the GSTM2 gene product into the kidney and to evaluate its potential therapeutic effect on anti-GBM-GN.MethodsThe human GSTM2 gene (hGSTM2) was transduced into mouse bone marrow-derived MSCs via a lentivirus vector to create a stable cell line (hGSTM2-MSC). The cultured hGSTM2-MSCs were treated with 0.5mM H2O2, and apoptotic cells were measured by terminal dUTP nick-end labeling (TUNEL) assay. The 129/svj mice, which were challenged with anti-GBM antibodies, were injected with 106 hGSTM2-MSCs via the tail vein. Expression of hGSTM2 and inflammatory cytokines in the kidney was assayed by quantitative PCR and western blotting. Renal function of mice was evaluated by monitoring proteinuria and levels of blood urea nitrogen (BUN), and renal pathological changes were analyzed by histochemistry. Immunohistochemical analysis was performed to measure inflammatory cell infiltration and renal cell apoptosis.ResultsMSCs transduced with hGSTM2 exhibited similar growth and differentiation properties to MSCs. hGSTM2-MSCs persistently expressed hGSTM2 and resisted H2O2-induced apoptosis. Upon injection into 129/svj mice, hGSTM2-MSCs migrated to the kidney and expressed hGSTM2. The anti-GBM-GN mice treated with hGSTM2-MSCs exhibited reduced proteinuria and BUN (58% and 59% reduction, respectively) and ameliorated renal pathological damage, compared with control mice. Mice injected with hGSTM2-MSCs showed alleviated renal inflammatory cell infiltration and reduced expression of chemokine (C-C motif) ligand 2 (CCL2), interleukin (IL)-1β and IL-6 (53%, 46% and 52% reduction, respectively), compared with controls. Moreover, hGSTM2-MSCs increased expression of renal superoxide dismutase and catalase, which may associate with detoxifying reactive oxygen species to prevent oxidative renal damage.ConclusionsOur data suggest that the enhanced protective effect of GSTM2-transduced MSCs against anti-GBM-GN might be associated with inhibition of oxidative stress-induced renal cell apoptosis and inflammation, through over-expression of hGSTM2 in mouse kidneys.

Molecular Alterations in the Cerebellum of Sporadic Creutzfeldt–Jakob Disease Subtypes with DJ-1 as a Key Regulator of Oxidative Stress

“…DJ-1 also regulates the glutathione activity by increasing the expression of the enzyme GCL which is the first enzyme of the pathway of cellular GSH biosynthesis [97]. Glutathione S-transferase Mu 2 is a class of enzymes which protects the cells by detoxifying electrophilic compounds like carcinogens, drug residues, environmental toxins and toxic products of oxidative stress [98]. GST Mu 2 expression was found significantly increased both at transcriptional and translational level in MM1 and VV2 subtypes of sCJD in present study.…”

Assessment of Curative Effect of Using Stem Cells on the Control of Pesticide Damaging Effect on Liver

“…We detected PKH26-labeled BM-MSCs in the injured liver tissue of recipient mice. (Quintanilha et al, 2013;Li et al, 2014).…”

Anti-inflammatory effects of alpinone 3-acetate from Alpinia japonica seeds

“…USP2a was shown to negatively regulate IL-1b-and virusinduced NF-jB activation by deubiquitinating TRAF6 [25]. In contrast, glutathione S-transferase was differentially upregulated by hydrocortisone, and reportedly ameliorated anti-glomerular basement membrane antibodyinduced glomerulonephritis by inhibiting oxidation and inflammation in glutathione S-transferase Mu 2-transduced mesenchymal stem cells [26]. In addition, the CXC chemokine IL8 is a major mediator of the inflammatory responses [27].…”