The global shortage of corneal endothelial graft tissue necessitates the exploration of alternative therapeutic strategies. Rho-associated protein kinase inhibitors (ROCKi), recognized for their regenerative potential in cardiology, oncology, and neurology, have shown promise in corneal endothelial regeneration. This study investigates the repurposing potential of additional ROCKi compounds. Through screening a self-assembled library of ROCKi on B4G12 corneal endothelial cells, we evaluated their dose-dependent effects on proliferation, migration, and toxicity using live-cell imaging. Nine ROCKi candidates significantly enhanced B4G12 proliferation compared to the basal growth rate. These candidates were further assessed for their potential to accelerate wound closure as another indicator for tissue regeneration capacity, with most demonstrating notable efficacy. To assess the potential impact of candidate ROCKi on key corneal endothelial cell markers related to cell proliferation, leaky tight junctions and ion efflux capacity, we analyzed the protein expression of cyclin E1, CDK2, p16, ZO-1 and Na+/K+-ATPase, respectively. Immunocytochemistry and western blot analysis confirmed the preservation of corneal endothelial markers post-treatment with ROCKi hits. However, notable cytoplasm enlargement and nuclear fragmentation were detected after the treatment with SR-3677 and Thiazovivin, indicating possible cellular stress. In compared parameters, Chroman-1 at a concentration of 10 nM outperformed other ROCKi, requiring significantly 1000-fold lower effective concentration than established ROCKi Y-27632 and Fasudil. Altogether, this study underscores the potential of repurposing ROCKi for treating corneal endothelial dysfunctions, offering a viable alternative to conventional grafting methods, and highlights Chroman-1 as a promising candidate structure for hit-to-lead development.

• Klíčová slova
• Chroman-1, Corneal endothelial regeneration, Drug repurposing, ROCK inhibitors, Small molecule screening,
• MeSH
• buněčné linie MeSH
• endoteliální buňky účinky léků   MeSH
• inhibitory proteinkinas * farmakologie   MeSH
• kinázy asociované s rho * antagonisté a inhibitory   metabolismus   MeSH
• lidé MeSH
• pohyb buněk účinky léků   MeSH
• přehodnocení terapeutických indikací léčivého přípravku MeSH
• proliferace buněk * účinky léků   MeSH
• regenerace * účinky léků   MeSH
• rohovkový endotel * účinky léků   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• inhibitory proteinkinas * MeSH
• kinázy asociované s rho * MeSH

The role of platelets in hemostasis may be influenced by alteration of the platelet redox state-the presence of antioxidants and the formation of reactive oxygen and nitrogen species. We investigated the effects of two antioxidants, resveratrol and trolox, on platelet activation. Trolox and resveratrol inhibited aggregation of washed platelets and platelet-rich plasma activated by ADP, collagen, and thrombin receptor-activating peptide. Resveratrol was a more effective agent in reducing platelet static and dynamic adhesion in comparison with trolox. The antioxidant capacity of resveratrol was, however, the same as that of trolox. After incubation of platelets with antioxidants, the resveratrol intraplatelet concentration was about five times lower than the intracellular concentration of trolox. Although both antioxidants comparably lowered hydroxyl radical and malondialdehyde production in platelets stimulated with collagen, TxB(2) levels were decreased by resveratrol much more effectively than by trolox. Cyclooxygenase 1 was inhibited by resveratrol and not by trolox. Our data indicate that antioxidants, apart from nonspecific redox or radical-quenching mechanisms, inhibit platelet activation also by specific interaction with target proteins. The results also show the importance of studying platelet activation under conditions of real blood flow in contact with reactive surfaces, e.g., using dynamic adhesion experiments.

• MeSH
• agregace trombocytů účinky léků   fyziologie   MeSH
• antioxidancia farmakologie   MeSH
• buněčná adheze účinky léků   fyziologie   MeSH
• chromany (dihydrobenzopyrany) farmakologie   MeSH
• cyklooxygenasa 1 metabolismus   MeSH
• hemostáza účinky léků   MeSH
• kolagen farmakologie   MeSH
• kultivované buňky MeSH
• lidé MeSH
• malondialdehyd metabolismus   MeSH
• resveratrol MeSH
• stilbeny farmakologie   MeSH
• thromboxan B2 metabolismus   MeSH
• trombocyty účinky léků   fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid MeSH Prohlížeč
• antioxidancia MeSH
• chromany (dihydrobenzopyrany) MeSH
• cyklooxygenasa 1 MeSH
• kolagen MeSH
• malondialdehyd MeSH
• resveratrol MeSH
• stilbeny MeSH
• thromboxan B2 MeSH

Current diagnostic techniques of prostate cancer cannot efficiently distinguish the latent and low-risk forms from the high-risk significant forms of prostate cancer. Caveolin-1 (Cav-1), except other functions, plays an important role in cell transformation and the process of tumorigenesis. Furthermore, Cav-1 is involved in metastatic processes. It has also been shown that Cav-1 expression is induced under stress conditions, such as oxidative stress. The present study focused on the determination of prognostic markers of aggressive (high-grade) forms of prostate cancer. We determined serum Cav-1 and serum markers of antioxidant activity-glutathione (GSH), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Trolox equivalent antioxidant capacity (TEAC), ferric-reducing antioxidant power (FRAP), N,N-dimethyl-1,4-diaminobenzene (DMPD), free radicals method (FRK) and blue chromium peroxide (Cro) in 97 serum samples (82 prostate cancer patients and 15 controls). We found insignificant differences in Cav-1 between the sera of patients and controls (5.69 in the cancer group vs. 5.42 ng/ml in the control group). However, we found a significant (p<0.004) 2.8-fold elevation of Cav-1 in high tumour stages (TNM T4) compared to lower stages and a significant positive association with histological grading (r=0.29, p=0.028). We also found that in patients with high serum Cav-1 the antioxidant capacity of the body is reduced. These findings indicate that Cav-1 may be an interesting tool for the prediction of disease burden.

• MeSH
• antioxidancia metabolismus   MeSH
• bifenylové sloučeniny krev   MeSH
• chromany (dihydrobenzopyrany) krev   metabolismus   MeSH
• glutathion krev   MeSH
• kaveolin 1 krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové biomarkery krev   MeSH
• nádory prostaty krev   MeSH
• oxidační stres MeSH
• peroxidy krev   MeSH
• pikráty krev   MeSH
• senioři MeSH
• sloučeniny chromu krev   MeSH
• volné radikály krev   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 1,1-diphenyl-2-picrylhydrazyl MeSH Prohlížeč
• 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid MeSH Prohlížeč
• antioxidancia MeSH
• bifenylové sloučeniny MeSH
• chromany (dihydrobenzopyrany) MeSH
• chromium peroxide MeSH Prohlížeč
• glutathion MeSH
• kaveolin 1 MeSH
• nádorové biomarkery MeSH
• peroxidy MeSH
• pikráty MeSH
• sloučeniny chromu MeSH
• volné radikály MeSH