Carbenoxolone mitigates extensive fibrosis formation in PLP-induced EAE model and multiple sclerosis serum-exposed pericyte culture.

Introduction: Multiple sclerosis (MS) is one of the most common causes of disability in young adults. Nearly, 85% of MS cases start with attacks and remissions, classified as relapsing-remitting multiple sclerosis (RRMS). With repeating attacks, MS causes brain-spinal cord atrophy and enhanced disability as disease progresses. PLP-induced EAE is one of the most established models for pathophysiology and treatment of RRMS. Recent studies demonstrated the possible role of pericytes in perivascular and intra-lesional fibrosis in PLP-induced EAE, whose importance remains elusive. Hence, we have investigated the possible role of pericytes in fibrosis formation and amelioration with a hemichannel blocker, Carbenoxolone (CBX). Methods: PLP-induced experimental autoimmune encephalitis (EAE) model is used and the effect of CBX is investigated. Clinical scores were recorded and followed. Perivascular Collagen 1 and 3 accumulations were demonstrated as markers of fibrosis in the spinal cord. To delineate the role of pericytes, human brain vascular pericytes (HBVP) were incubated with the sera of MS patients to induce in-vitro MS model and the fibrosis formation was investigated. Results: In the PLP induced in-vivo model, both intracerebroventricular and intraperitoneal CBX have significantly mitigated the disease progression followed by clinical scores, demyelination, and fibrosis. Moreover, CBX significantly mitigated MS-serum-induced fibrosis in the HBVP cell culture. Discussion: The study demonstrated two important findings. First, CBX decreases fibrosis formation in both in-vivo and in-vitro MS models. Secondly, it improves neurological scores and decreases demyelination in the EAE model. Therefore, CBX can be potential novel therapeutic option in treating Multiple Sclerosis.

Protective Effects of Infliximab Against Kanamycin-Induced Ototoxicity in Rats.

HYPOTHESIS: To examine the protective effects of infliximab (INF) against kanamycin (KM)-induced hearing loss. BACKGROUND: Tumor necrosis factor α blockers can reduce cellular inflammatory reactions and decrease cell death. METHODS: Thirty-six rats with normal hearing were randomly divided into six groups. The first group was injected with 400 mg/kg KM intramuscularly (IM), the second group with 7 mg/kg INF intraperitoneally (IP) and 400 mg/kg KM IM, the third group with 7 mg/kg INF IP and 200 mg/kg KM IM, and the fourth group with 1 mg/kg 6-methylprednisolone (MP) IP and 400 mg/kg KM IM. Group 5 was injected with 1 mg/kg MP IP and 200 mg/kg KM IM, and group 6 with saline IP once. Auditory brain-stem response (ABR) for hearing thresholds was performed on days 7 and 14. From the frozen sections of the cochlea, the area of the stria vascularis, the number of neurons in the spiral ganglion, the fluorescence intensity of hair cells (FIHC), postsynaptic density (PSD), and presynaptic ribbons (PSRs) were calculated. RESULTS: The KM-induced increase in hearing thresholds was detected on the 14th day. Hearing was only preserved in the group treated with INF after low-dose KM exposure but not in the groups that received high-dose KM. The FIHC, excitatory PSD, and PSR were preserved only in the INF-treated group after half-dose KM exposure. In MP groups, FIHC, excitatory PSD, and PSR were significantly lower than in the control group. CONCLUSIONS: Our results support that tumor necrosis factor-based inflammation may play a role in the ototoxicity mechanism.

The sex-dependent anti-depressant-like effects of zeatin in rat behavioral despair model as a candidate A2A receptor ligand.

Zeatin, an adenine-derivative cytokinin has well-established functions in plants. It is also suggested to activate A2A receptors in animals, however, there is limited knowledge of its effects. The main objective of this study is to evaluate the possible effects of zeatin on depression, and our hypothesis is that zeatin might induce an anti-depressant effect via A2A receptor-linked pathways. The forced swim test was used to create a depression-like model on female and male rats. A balanced zeatin isomer mixture (80 % trans-zeatin (tZ), 20 % cis-zeatin (cZ)) was administered intraperitoneally to analyze the effects. Caffeine with a suboptimal dose (2 mg/kg) was used as a known ligand of A2A receptor. Finally, a molecular docking study was also implemented to compare caffeine and tZ in the ligand binding site of A2A receptor. We demonstrate that (1) there is a clear sex-dependent difference in the susceptibility to depression-like symptoms, where female rats in the metestrus phase display higher depressive-like behavior and lower responses to the anti-depressant-like effects of pharmacological applications; (2) 10 mg/kg zeatin exerts an anti-depressant-like effect for both females and males without affecting locomotor activity; (3) 8 mg/kg tZ alone replicates this effect for both sexes, (4) the effect of zeatin is also differential for either sex and (5) the similar effect of caffeine and zeatin implies that the effect might be exerted via A2A receptor mediated pathways. Computational analysis further yielded similar binding patterns for both ligands. In conclusion, zeatin might have a potential therapeutic use in depression, acting via adenosinergic pathways.