The Flavonoid Naringenin Alleviates Collagen-Induced Arthritis through Curbing the Migration and Polarization of CD4+ T Lymphocyte Driven by Regulating Mitochondrial Fission

Abstract

Rheumatoid arthritis (RA) is a progressive autoimmune disease. Due to local infiltration and damage to the joints, activated CD4+ T cells play a crucial role in the progression of RA. However, the exact regulatory mechanisms are perplexing, which makes the effective management of RA frustrating. This study aimed to investigate the effect of mitochondria fission on the polarization and migration of CD4+ T cells as well as the regulatory mechanism of NAR, so as to provide enlightenment on therapeutic targets and novel strategies for the treatment of RA. In this study, a collagen-induced arthritis (CIA) model was established, and rats were randomly given saline or naringenin (NAR, 10 mg/kg, 20 mg/kg, 50 mg/kg, i.p.) once a day, before being euthanized on the 42nd day of primary immunization. The pain-like behavior, articular index scores, account of synovial-infiltrated CD4+ T cells, and inflammatory factors were investigated in each group. In vitro, spleen CD4+ T lymphocytes were derived from each group. In addition, mitochondrial division inhibitor 1 (Mdivi-1) or NAR was added to the cell medium containing C-X-C motif chemokine ligand 12 (CXCL12) in order to induce CD4+ T lymphocytes, respectively. The polarization capacity of CD4+ T cells was evaluated through the immunofluorescence intensity of the F-actin and myosin light chain phosphorylated at Ser19 (pMLC S19), and the mitochondrial distribution was determined by co-localization analysis of the translocase of outer mitochondrial membrane 20 (TOM20, the mitochondrial marker) and intercellular adhesion molecule 1 (ICAM1, the uropod marker). The mitochondrial fission was investigated by detecting dynamin-related protein 1 (Drp1) and mitochondrial fission protein 1 (Fis1) using Western blot and immunofluorescence. This study revealed that high-dose NAR (50 mg/kg, i.p.) alleviated pain-like behavior and articular index scores, reduced the serum level of interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α), and accounted for CD4+ T lymphocytes that infiltrated into the synovial membrane of the CIA group. Meanwhile, NAR (50 mg/kg, i.p.) suppressed the polarization of spleen CD4+ T lymphocytes, reduced the redistribution of mitochondria in the uropod, and inhibited the expression of Drp1 and Fis1 in the CIA model. Furthermore, the in vitro experiments confirmed that NAR reduced mitochondrial fission, which in turn inhibited the CXCL12-induced polarization and migration of CD4+ T lymphocytes. Our results demonstrated that the flavonoid NAR was a promising drug for the treatment of RA, which could effectively interfere with mitochondrial fission, thus inhibiting the polarization and migration of CD4+ T cells in the synovial membrane.

Rheumatoid arthritis molecular targets and their importance to flavonoid‐based therapy

“…Moreover, delphinidin showed ability to counteract the cytokine‐induced immune response in RA by reducing pro‐inflammatory cytokine expression in Jurkat T lymphocytes, through the inhibition of NF‐κB/p65 pathway 96 . Similar effects were shown for naringenin , where the reduction of CD4+ lymphocytes infiltration in the synovial membrane 158 and the reduction of the expression of costimulatory molecules, the reduction of the antigen‐specific T cell activation, and the reduction of MAPK and NF‐κB signaling activation in DC were signs of its immunomodulatory properties. These effects corroborate the observed limited proliferation of T cells (Th1 and Th17) and reduction of the severity of arthritis in a CIA mouse model treated with this flavonoid 157 …”

“…Furthermore, a significant number of flavonoids also showed ability to modulate several other pro‐inflammatory cytokines in the context of RA in both in vitro and in vivo models (Table 2). For instance, apigenin 81 and naringenin 158 reduced the secretion of pro‐inflammatory cytokines in the serum of mice subjected to collagen‐induced arthritis (CIA), which delayed the onset and reduced the severity of arthritis in the animals. This is similar to what it is described by Mohammad‐Shahi et al 94 and by Hu et al 118 for daidzein and genistein , respectively.…”

Intramammary Infusion of Micronised Purified Flavonoid Fraction (MPFF) in Mastitis-Diagnosed Dairy Cows Naturally Infected by Staphylococcus spp. in the Late Lactation

“…The inflammatory process mediated by flavonoids inhibits signaling transduction pathways and secretory processes [ 87 ] together with a decreased cell proliferation and a lower release of pro-inflammatory cytokines [ 48 , 88 ]. Moreover, the flavonoids inhibit the cell maturation by suppressing the response of CD4 + T cells and diminishing several IL-family effects through blocking IL-cytokine and IgE receptors [ 88 , 89 ]. Therefore, MPFF might also activate or inhibit other receptors located in different tissues such as the mammary gland; however, this has yet to be described.…”

Potential Beneficial Effects of Naringin and Naringenin on Long COVID—A Review of the Literature

“…Naringenin modulates the immune system’s response to autoantigens by influencing the growth and specialization of T cells, as well as cytokine signaling [ 98 , 99 ]. Naringenin effectively alleviates symptoms of rheumatoid arthritis in rats by modulating lymphocyte polarization, primarily through the reduction in T helper (Th)1 and Th17 cell differentiation and the reduction in IL-6 and TNF-α levels [ 99 , 100 ]. Similarly, naringenin alleviates symptoms of multiple sclerosis and systemic lupus erythematosus by inhibiting the growth and specialization of harmful pro-inflammatory T cells while maintaining the differentiation of anti-inflammatory subgroups [ 101 , 102 ].…”