20.500.12556/RUL-179136 Signaliziranje s kalcijem v povezavi s kanali TRPV4 v epitelu leče pri bolnikih z različnimi stopnjami sive mrene Calcium signaling in relation to TRPV4 channels in the lens epithelium of patients with different degrees of cataracts Očesna leča je prozoren epitelijsko-fibrilarni organ, katerega prosojnost je med drugim odvisna od uravnavanja znotrajcelične koncentracije kalcija ([Ca²⁺]ᵢ). Motnje v signalizaciji kalcijevih ionov (Ca²⁺) so povezane z razvojem sive mrene (katarakte), ki je najpogostejši vzrok za izgubo vida. Med pomembnimi uravnalci homeostaze Ca²⁺ v leči so ionski kanali iz družine TRP, zlasti TRPV4, ki deluje kot mehanosenzitiven in osmotsko odziven kanal, odgovoren za vstop Ca²⁺ in vzdrževanje celične funkcionalnosti. V raziskavi je bila analizirana aktivnost TRPV4 kanalov in njihov vpliv na Ca²⁺ signalizacijo glede na napredovanje katarakte. Pooperativni vzorci lečnega epitela so bili pridobljeni pri bolnikih z različnimi stopnjami bolezni na Očesni kliniki UKC Ljubljana. Celice so bile obarvane s fluorescentnim kalcijevim indikatorjem Fura2, spremembe [Ca²⁺]ᵢ pa spremljane po mehanski stimulaciji in ob uporabi TRPV4 antagonista HC-067047. Rezultati kvantitativne analize so pokazali, da mehanska stimulacija sproži hiter, prehoden dvig [Ca²⁺]ᵢ, ki se širi medcelično. Uporaba TRPV4 antagonista je zmanjšala amplitudo ter upočasnila širjenje Ca²⁺ vala, kar podpira vlogo TRPV4 pri vzdrževanju učinkovitih Ca²⁺ odzivov in medcelične komunikacije. Učinek TRPV4 antagonista je bil izrazitejši v zgodnejših stadijih bolezni, medtem ko je bil pri napredovalih kataraktah manj izrazit. Kljub temu je treba poudariti, da je bil vzorec omejen, razlike med skupinami pa relativno blage, zato je rezultate treba interpretirati previdno. Opaženi trendi lahko odražajo spremembe v Ca²⁺ signalizaciji, povezane z napredovanjem katarakte, vendar za zanesljivejše sklepe o spremembah funkcionalnosti TRPV4 kanalov potrebujemo dodatne raziskave na večjem številu vzorcev. The ocular lens is a transparent epithelial–fibrous organ whose transparency depends, among other factors, on the regulation of intracellular calcium concentration ([Ca²⁺]ᵢ). Disruptions in calcium ion (Ca²⁺) signaling are associated with the development of cataract, the most common cause of vision loss. Among the key regulators of calcium homeostasis in the lens are TRP family ion channels, particularly TRPV4, which functions as a mechanosensitive and osmosensitive channel responsible for Ca²⁺ influx and the maintenance of cellular functionality. This study investigated TRPV4 channel activity and its role in Ca²⁺ signaling during cataract progression. Postoperative human lens epithelial samples were obtained from patients at different disease stages at the Eye Hospital, University Medical Centre Ljubljana. Cells were stained with the fluorescent calcium indicator Fura-2, and changes in [Ca²⁺]ᵢ were monitored following mechanical stimulation and after application of the TRPV4 antagonist HC-067047. Quantitative analysis showed that mechanical stimulation triggered a rapid, transient increase in [Ca²⁺]ᵢ that propagated intercellularly. Application of the TRPV4 antagonist reduced the amplitude and slowed the propagation of the Ca²⁺ signal, demonstrating that TRPV4 contributes to maintaining effective Ca²⁺ responses and intercellular communication. The effect of the TRPV4 antagonist was more pronounced in the earlier stages of the disease, whereas this effect was less evident in advanced stages of cataract. However, it should be emphasized that the sample size was limited and the observed differences were relatively subtle; therefore, the results should be interpreted with caution. The observed trends may reflect changes in Ca²⁺ signaling associated with cataract progression, but further studies with a larger number of samples are required to draw firm conclusions regarding changes in TRPV4 channel functionality. lečni epitelij TRPV4 TRPV4 antagonist kalcijeva signalizacija siva mrena mehanska stimulacija lens epithelium TRPV4 TRPV4 antagonist calcium signaling cataract mechanical stimulation true false false [B. Kinkela] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-02-06 07:15:55 2026-02-06 07:16:01 2026-02-12 04:35:31 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 617.741-004.1(043.2) 278878 267587587 56379.pdf 56379.pdf 1 7A1B4B3D5ADEEC2A6D518EE0A2FDCBD4 7f9f747e81367486218678b6f7bfd6980590bd18f53217950c2250d539e848cb 3502a884-0323-11f1-a1ba-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=227347 Biotehniška fakulteta 0 0 0