Protective Effect of PBCA Nanoparticles Loaded with Thymulin Against the Relapsing-Remitting Form of Experimental Autoimmune Encephalomyelitis in Mice

Abstract

Relapsing–remitting experimental autoimmune encephalomyelitis (rEAE) in mice is a model that closely resembles relapsing–remitting multiple sclerosis in humans. This study aims to investigate a new approach to modulation of the inflammatory response in rEAE mice using a thymic peptide thymulin bound to polybutylcyanoacrylate (PBCA) nanoparticles. PBCA nanoparticles were used to prolong the presence of thymulin in the blood. Cytokine levels in blood were measured by ELISA; NF-κB and SAPK/JNK cascade activation, as well as Hsp72 and p53 protein expression, were measured by Western blotting. Animal health statuses were estimated using severity scores. Results showed that the cytokine response in rEAE was multi-staged: an early phase was accompanied by an increase in plasma interferon-γ, while the interleukin (IL)-17 response was markedly increased at a later stage. The stages were attributed to rEAE induction and maintenance phases. Thymulin significantly alleviated symptoms of rEAE and lowered plasma cytokine levels both in early and later stages of rEAE, and decreased NF-κB and SAPK/JNK cascade activation. Thymulin modulated NF-kappaB pathway activity via site-specific phosphorylation of RelA/p65 protein (at Ser276 and Ser536). The effect of nanoparticle-bound thymulin was more pronounced than the effect of free thymulin. Therefore, PBCA–thymulin can be considered a prospective treatment for this pathology.

Thymulin and peroxiredoxin 6 have protective effects against streptozotocin-induced type 1 diabetes in mice

“…However, the most striking effect of thymulin was the normalization of JNK signaling. These results are consistent with data that thymulin is an important modulator in inflammations of various etiology, such as septic-type inflammations 10 or autoimmune diseases, 13 , 14 and with the findings on the possible role of thymulin in response to oxidative damage. 25 The study have some limitations.…”

“…The immunomodulatory and anti-inflammatory effect of thymulin was previously shown in mice with septic-type inflammation 11 , 12 and in mice with experimental autoimmune encephalomyelitis. 13 , 14 Furthermore, our previous study showed beneficial effects of PRDX6 in RIN-m5F β-cells in vitro 15 and of thymulin in mice with diabetes induced by alloxan administration in vivo. 16 , 17 In addition, a significant stimulatory effect of PRDX6 on the insulin-producing activity of pancreatic β-cells was demonstrated, and, most importantly, the effect of PRDX6 was detected during culturing the cells under both normal and diabetes-modeling conditions.…”

“…16 Western blot analysis Splenic cells (10 8 cells per sample) were lysed, and the total protein measurement using the Bradford solution (Sigma, USA), the PAGE electrophoresis using protein molecular weight (MW) marker (Thermo Scientific, USA), the protein transfer from the gel onto a nitrocellulose membrane (GE Healthcare, Amersham, UK), and the blockade with 5% w/v nonfat dry milk in TBS/Tween 20 were performed as reported earlier. 14 After blocking, membranes were incubated for 2 h with antibodies against the following murine proteins: anti-phospho-NF-κB p65 (at Ser 536) antibody (cat. number 3031, Cell Signaling Technology, Danvers, MA, USA), rabbit phospho-stress-activated protein kinase/JNK (SAPK/JNK) antibody (Cell Signaling Technology), and rabbit polyclonal antibody to HSP90a (Enzo Life Sciences, USA) and washed.…”

Promising Nanotechnology Approaches in Treatment of Autoimmune Diseases of Central Nervous System

“…Lunin et al [106] developed poly(butylcyanoacrylate) (PBCA) nanoparticles, in order to deliver the thymic peptide thymulin and prolong its presence in the blood of mice with relapsing/remitting EAE (rEAE). The increase in the blood content of thymulin, which is depleted with age, and the prolongation of its blood half-life, alone or in combination with other treatments, may be a prospective strategy for treatment of chronic inflammatory conditions, such as MS. More analytically, as revealed by the results, thymulin significantly decreased symptoms of rEAE and lowered plasma cytokine levels, both in early and in later stages of rEAE, as well as decreased the NF-κB and stress-activated protein kinase/Jun amino-terminal kinase (SAPK/JNK) cascade activation, as confirmed by ELISA measurements of cytokine levels in the blood.…”

NF-κB and Disease

“…Thymulin and other thymic peptides, in fact, are characterized by their very low half-life in plasma (less than 10 min) and in vivo data show that orally administered poly-butylcyanoacrylate (PBCA) nanoparticles containing the thymic hormone thymopentin had similar effects to intravenous thymopentin, but with enhanced bioavailability due to the nanoparticles [6].…”