[en] Purpose: In this study, we investigated the biochemical pharmacology of pirenoxine (PRX) and catalin under in vitro selenite/calcium- and ultraviolet (UV)-induced lens protein turbidity challenges. The systemic effects of catalin were determined using a selenite-induced cataractogenesis rat model.
Methods: In vitro cataractogenesis assay systems (including UVB/C photo-oxidation of lens crystallins, calpain-induced proteolysis, and selenite/calcium-induced turbidity of lens crystallin solutions) were used to screen the activity of PRX and catalin eye drop solutions. Turbidity was identified as the optical density measured using spectroscopy at 405 nm. We also determined the in vivo effects of catalin on cataract severity in a selenite-induced cataract rat model. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE) was applied to analyze the integrity of crystallin samples.
Results: PRX at 1,000 μM significantly delayed UVC-induced turbidity formation compared to controls after 4 h of UVC exposure (p<0.05), but not in groups incubated with PRX concentrations of <1,000 μM. Results were further confirmed by SDS–PAGE. The absolute γ-crystallin turbidity induced by 4 h of UVC exposure was ameliorated in the presence of catalin equivalent to 1~100 μM PRX in a concentration-dependent manner. Samples with catalin-formulated vehicle only (CataV) and those containing PRX equivalent to 100 μM had a similar protective effect after 4 h of UVC exposure compared to the controls (p<0.05). PRX at 0.03, 0.1, and 0.3 μM significantly delayed 10 mM selenite- and calcium-induced turbidity formation compared to controls on days 0~4 (p<0.05). Catalin (equivalent to 32, 80, and 100 μM PRX) had an initial protective effect against selenite-induced lens protein turbidity on day 1 (p<0.05). Subcutaneous pretreatment with catalin (5 mg/kg) also statistically decreased the mean cataract scores in selenite-induced cataract rats on post-induction day 3 compared to the controls (1.3±0.2 versus 2.4±0.4; p<0.05). However, catalin (equivalent to up to 100 μM PRX) did not inhibit calpain-induced proteolysis activated by calcium, and neither did 100 μM PRX.
Conclusions: PRX at micromolar levels ameliorated selenite- and calcium-induced lens protein turbidity but required millimolar levels to protect against UVC irradiation. The observed inhibition of UVC-induced turbidity of lens crystallins by catalin at micromolar concentrations may have been a result of the catalin-formulated vehicle. Transient protection by catalin against selenite-induced turbidity of crystallin solutions in vitro was supported by the ameliorated cataract scores in the early stage of cataractogenesis in vivo by subcutaneously administered catalin. PRX could not inhibit calpain-induced proteolysis activated by calcium or catalin itself, and may be detrimental to crystallins under UVB exposure. Further studies on formulation modifications of catalin and recommended doses of PRX to optimize clinical efficacy by cataract type are warranted.
Pascolini D, Mariotti SP, Pokharel GP, Pararajasegaram R, Etya'ale D., Negrel AD, Resnikoff S. 2002 global update of available data on visual impairment: a compilation of population-based prevalence studies. Ophthalmic Epidemiol 2004; 11:67-115.
Resnikoff S, Pascolini D, Etya'ale D, Kocur I, Pararajasegaram R, Pokharel GP, Mariotti SP. Global data on visual impairment in the year 2002. Bull World Health Organ 2004; 82:844-51.
Frick KD, Foster A. The magnitude and cost of global blindness: an increasing problem that can be alleviated. Am J Ophthalmol 2003; 135:471-6.
Ogino S, Tojo H, Fujishige I, Katumori Y. Biochemical studies on cataract. IX. Contribution to the histopathology of cataract caused by various quinoid substances. Am J Ophthalmol 1957; 44:94-105.
Ogino S, Yasukura K. Biochemical studies on cataract. VI. Production of cataracts in guinea pigs with dinitrophenol. Am J Ophthalmol 1957; 43:936-46.
Kociecki J, Zalecki K, Wasiewicz-Rager J, Pecold K. Evaluation of effectiveness of Catalin eyedrops in patients with presenile and senile cataract Polish. Klin Oczna 2004; 106:778-82.
Chandorkar AG, Albal MV, Bulakh PM, Jain PK. Hypoglycaemic effect of 'Catalin' an anti-cataract agent in rabbits, (a preliminary study). Indian J Ophthalmol 1978; 26:6-8.
Bulakh PM, Chandorkar AG, Balsara JJ, Ranade SM, Albal MV. Effect of 'catalin' an anticataract agent on alloxan induced hyperglycaemia and diabetic cataract in rats. Indian J Ophthalmol 1980; 28:1-3.
Sekimoto M, Imanaka Y, Kitano N, Ishizaki T, Takahashi O. Why are physicians not persuaded by scientific evidence? A grounded theory interview study. BMC Health Serv Res 2006; 6:92.
Murata T. Double blind study on the effectiveness of a pirenoxine eye drop to senile cataract in Japanese. Nippon Ganka Kiyo 1980; 31:1217-22.
Dilsiz N, Olcucu A, Atas M. Determination of calcium, sodium, potassium and magnesium concentrations in human senile cataractous lenses. Cell Biochem Funct 2000; 18:259-62.
Garner MH, Kuszak JR. Cations, oxidants, light as causative agents in senile cataracts. P R Health Sci J 1993; 12:115-22.
Liao JH, Chen CS, Maher TJ, Liu CY, Lin MH, Wu TH, Wu SH. Astaxanthin interacts with selenite and attenuates selenite-induced cataractogenesis. Chem Res Toxicol 2009; 22:518-25.
Liao JH, Chen CS, Hu CC, Chen WT, Wang SP, Lin IL, Huang YH, Tsai MH, Wu TH, Huang FY, Wu SH. Ditopic complexation of selenite anions or calcium cations by pirenoxine: an implication for anti-cataractogenesis. Inorg Chem 2011; 50:365-77.
Wu TH, Liao JH, Hou WC, Huang FY, Maher TJ, Hu CC. Astaxanthin protects against oxidative stress and calcium- induced porcine lens protein degradation. J Agric Food Chem 2006; 54:2418-23.
Ostádalová I, Babicky A, Obenberger J. Cataract induced by administration of a single dose of sodium selenite to suckling rats. Experientia 1978; 34:222-3.
Shearer TR, Anderson RS, Britton JL. Uptake and distribution of radioactive selenium in cataractous rat lens. Curr Eye Res 1982-1983; 2:561-4.
Gupta SK, Trivedi D, Srivastava S, Joshi S, Halder N, Verma SD. Lycopene attenuates oxidative stress induced experimental cataract development: an in vitro and in vivo study. Nutrition 2003; 19:794-9.
Dawczynski J, Winnefeld K, Konigsdorffer E, Augsten R, Blum M, Strobel J. Klin Monatsbl Augenheilkd 2006; 223:675-80. Selenium and cataract-risk factor or useful dietary supplement?
Tarwadi K, Agte V. Linkages of antioxidant, micronutrient, and socioeconomic status with the degree of oxidative stress and lens opacity in Indian cataract patients. Nutrition 2004; 20:261-7.
Ciaralli L, Giordano R, Costantini S, Sepe A, Cruciani F, Moramarco A, Antonelli B, Balacco-Gabrieli C. Element concentrations and cataract: an experimental animal model. J Trace Elem Med Biol 2001; 14:205-9.
Rasi V, Costantini S, Moramarco A, Giordano R, Giustolisi R, Balacco Gabrieli C. Inorganic element concentrations in cataractous human lenses. Ann Ophthalmol 1992; 24:459-64.
David LL, Shearer TR. Beta-crystallins insolubilized by calpain II in vitro contain cleavage sites similar to beta-crystallins insolubilized during cataract. FEBS Lett 1993; 324:265-70.
Fukiage C, Azuma M, Nakamura Y, Tamada Y, Shearer TR. Calpain-induced light scattering by crystallins from three rodent species. Exp Eye Res 1997; 65:757-70.
Spector A, Roy D. Disulfide-linked high molecular weight protein associated with human cataract. Proc Natl Acad Sci USA 1978; 75:3244-8.
Lou MF, Dickerson JE Jr. Protein-thiol mixed disulfides in human lens. Exp Eye Res 1992; 55:889-96.
Wu K, Kojima M, Shui YB, Sasaki K, Hockwin O. In vitro UVB effect on lens protein solutions. Ophthalmic Res 1997; 29:75-82.
McCarty CA, Taylor HR. Recent developments in vision research: light damage in cataract. Invest Ophthalmol Vis Sci 1996; 37:1720-3.
Zigman S. Environmental near-UV radiation and cataracts. Optom Vis Sci 1995; 72:899-901.
Sommerburg O, Ullrich O, Sitte N, von Zglinicki D, Siems W, Grune T. Dose- and wavelength-dependent oxidation of crystallins by UV light-selective recognition and degradation by the 20S proteasome. Free Radic Biol Med 1998; 24:1369-74.
Fujii N, Uchida H, Saito T. The damaging effect of UV-C irradiation on lens alpha-crystallin. Mol Vis 2004; 10:814-20.
Adelman R, Saul RL, Ames BN. Oxidative damage to DNA: relation to species metabolic rate and life span. Proc Natl Acad Sci USA 1988; 85:2706-8.
Lee JS, Liao JH, Wu SH, Chiou SH. alpha-Crystallin acting as a molecular chaperone against photodamage by UV irradiation. J Protein Chem 1997; 16:283-9.
Liao JH, Lee JS, Chiou SH. Distinct roles of alphaA- and alphaB-crystallins under thermal and UV stresses. Biochem Biophys Res Commun 2002; 295:854-61.
Andley UP, Clark BA. Spectroscopic studies on the photooxidation of calf-lens gamma-crystallin. Curr Eye Res 1988; 7:571-9.
Shih M, David LL, Lampi KJ, Ma H, Fukiage C, Azuma M, Shearer TR. Proteolysis by m-calpain enhances in vitro light scattering by crystallins from human and bovine lenses. Curr Eye Res 2001; 22:458-69.
Shearer TR, Ma H, Fukiage C, Azuma M. Selenite nuclear cataract: review of the model. Mol Vis 1997; 3:8.
Geraldine P, Sneha BB, Elanchezhian R, Ramesh E, Kalavathy CM, Kaliamurthy J, Thomas PA. Prevention of seleniteinduced cataractogenesis by acetyl-L-carnitine: an experimental study. Exp Eye Res 2006; 83:1340-9.
Rhodes JD, Sanderson J. The mechanisms of calcium homeostasis and signalling in the lens. Exp Eye Res 2009; 88:226-34.
Duncan G, Bushell AR. Ion analyses of human cataractous lenses. Exp Eye Res 1975; 20:223-30.
David LL, Shearer TR. Calcium-activated proteolysis in the lens nucleus during selenite cataractogenesis. Invest Ophthalmol Vis Sci 1984; 25:1275-83.
Sanderson J, Marcantonio JM, Duncan G. A human lens model of cortical cataract: Ca2+-induced protein loss, vimentin cleavage and opacification. Invest Ophthalmol Vis Sci 2000; 41:2255-61.
Marcantonio JM, Duncan G, Rink H. Calcium-induced opacification and loss of protein in the organ-cultured bovine lens. Exp Eye Res 1986; 42:617-30.
Sharma Y, Rao CM, Narasu ML, Rao SC, Somasundaram T, Gopalakrishna A, Balasubramanian D. Calcium ion binding to delta- and to beta-crystallins. The presence of the "EFhand" motif in delta-crystallin that aids in calcium ion binding. J Biol Chem 1989; 264:12794-9.
Nakamura Y, Fukiage C, Azuma M, Shearer TR. Oxidation enhances calpain-induced turbidity in young rat lenses. Curr Eye Res 1999; 19:33-40.
Chen CS, Wu SH, Wu YY, Fang JM, Wu TH. Properties of astaxanthin/Ca2+ complex formation in the deceleration of cis/trans isomerization. Org Lett 2007; 9:2985-8.
Davies MJ, Truscott RJ. Photo-oxidation of proteins and its role in cataractogenesis. J Photochem Photobiol B 2001; 63:114-25.
Brian G, Taylor H. Cataract blindness-challenges for the 21st century. Bull World Health Organ 2001; 79:249-56.
Skouri-Panet F, Bonnete F, Prat K, Bateman OA, Lubsen NH, Tardieu A. Lens crystallins and oxidation: the special case of gammaS. Biophys Chem 2001; 89:65-76.
Ciuffi M, Pisanello M, Pagliai G, Raimondi L, Franchi-Micheli S, Cantore M, Mazzetti L, Failli P. Antioxidant protection in cultured corneal cells and whole corneas submitted to UV-B exposure. J Photochem Photobiol B 2003; 71:59-68.
Bando M, Mikuni I, Obazawa H. Acceleration of calciuminduced aggregation of rat lens soluble protein by photosensitization with 8-methoxypsoralen and 3-hydroxy- L-kynurenine O-beta-glucoside. Exp Eye Res 1982; 34:953-60.
Ciuffi M, Neri S, Franchi-Micheli S, Failli P, Zilletti L, Moncelli MR, Guidelli R. Protective effect of pirenoxine and U74389F on induced lipid peroxidation in mammalian lenses. An in vitro, ex vivo and in vivo study. Exp Eye Res 1999; 68:347-59.
Cantore M, Siano S, Coronnello M, Mazzetti L, Franchi- Micheli S, Boldrini E, Ciuffi M, Failli P. Pirenoxine prevents oxidative effects of argon fluoride excimer laser irradiation in rabbit corneas: biochemical, histological and cytofluorimetric evaluations. J Photochem Photobiol B 2005; 78:35-42.
Angra SK, Mohan M, Saini JS. Medical therapy of cataract (evaluation of Catalin). Indian J Ophthalmol 1983; 31:5-8.
Ito Y, Nagai N, Cai H, Takeda M, Koizumi Y. Preventive effect of eye drops of liposomes containing disulfiram and cefmetazole on selenite-induced cataract in rat pups. J Oleo Sci 2006; 55:15-22.
Sakthivel M, Geraldine P, Thomas PA. Alterations in the lenticular protein profile in experimental selenite-induced cataractogenesis and prevention by ellagic acid. Graefes Arch Clin Exp Ophthalmol. 2011
Yan H, Harding JJ, Hui YN, Li MY. Decreased chaperone activity of alpha-crystallin in selenite cataract may result from selenite-induced aggregation. Eye (Lond) 2003; 17:637-45.
Tamada Y, Fukiage C, Mizutani K, Yamaguchi M, Nakamura Y, Azuma M, Shearer TR. Calpain inhibitor, SJA6017, reduces the rate of formation of selenite cataract in rats. Curr Eye Res 2001; 22:280-5.
