Combination of 5‐aminolevulinic acid and iron prevents skin fibrosis in murine sclerodermatous graft‐versus‐host disease

Abstract

Scleroderma or systemic sclerosis (SSc) is a clinically heterogeneous rheumatological autoimmune disease affecting the skin, internal organs and blood vessels. There is at present no effective treatment for this condition. Our study investigated the effects of 5-aminolevulinic acid (5-ALA), which is a precursor of haem synthesis, on graft-vs-host disease (GvHD)-induced SSc murine model. Lymphocytes were intravenously injected from donor mice (B10.D2) into recipient BALB/c mice (recombination-activating gene 2 (Rag-2)-null mice) deficient in mature T and B cells to induce sclerodermatous GvHD (scl-GvHD). To investigate the effect of 5-ALA on scl-GvHD, combination of 5-ALA and sodium ferrous citrate (SFC) was orally administered to the recipient mice for 9 weeks. 5-ALA/SFC treatment significantly reduced progressive inflammation and fibrosis in the skin and ears. Furthermore, 5-ALA/SFC suppressed mRNA expression of transforming growth factor-β, type I collagen and inflammatory cytokines. These results indicate that the 5-ALA/SFC combination treatment has a protective effect against tissue fibrosis and inflammation in a murine scl-GvHD-induced skin and ear inflammation and fibrosis. Furthermore, the efficacy of 5-ALA/SFC suggests important implications of HO-1 protective activity in autoimmune diseases, and therefore, 5-ALA/SFC may have promising clinical applications. These findings suggested that the 5-ALA/SFC treatment may be the potential strategies for SSc.

5‐ALA/SFC Attenuated Binge Alcohol–Induced Gut Leakiness and Inflammatory Liver Disease in HIV Transgenic Rats

“…Binge alcohol raised the expression of TNF‐α, CCL2, leptin, and leptin receptor in rat liver in accordance with a previous report (Banerjee et al., ), whereas the administration of 5‐ALA/SFC suppresses the increased expression of these pro‐inflammatory molecules in rat liver (Fig. A ); this is consistent with our previous reports using other inflammatory animal models and 5‐ALA/SFC treatment (Hou et al., ; Li et al., ; Liu et al., ).…”

“…The quantification of neutrophils was based on the segmented morphology of the nucleus followed by staining with the Naphthol AS‐D Chloroacetate Esterase Staining Kit (Muto Pure Chemicals, Co. Ltd., Tokyo, Japan). Quantifications were performed using WinRoof V.6.1 software (Mitani Corporation, Tokyo, Japan) as described previously (Liu et al., ).…”

“…Total RNA isolation and quantitative real‐time RT‐PCR were performed as described previously (Liu et al., ). Primer sequences and probes used in this study are described in Table .…”

“…Western blot analysis was performed as described previously (Liu et al., ). In brief, proteins from liver were homogenized with RIPA lysis buffer (Wako, Osaka, Japan) containing phosphatase inhibitor cocktail and protease inhibitor cocktail.…”

5-Aminolevulinic acid combined with sodium ferrous citrate (5-ALA/SFC) ameliorated liver injury in a murine acute graft-versus-host disease model by reducing inflammation responses through PGC1-α activation

“…Growing evidence supports the notion that 5-ALA combined with sodium ferrous citrate (5-ALA/SFC) exerts antioxidant, anti-inflammatory and anti-fibrotic properties (8)(9)(10)(11). In addition, 5-ALA/ SFC has been used to treat a variety of animal models, including inflammatory disease, transplantation, autoimmune disease and sclerodermatous issues induced by chronic GvHD, which may explain its antiinflammatory, immunoregulation and cytoprotective properties exerted via the upregulation of heme oxygenase (HO)-1 expression and release of heme metabolites (12,13).…”