Cataractous Rat Lens Research Articles

Delaying Effect of Polyherbal Formulation on Cataract in STZ-NIC-induced Diabetic Wistar Rats

The aim of the study was to evaluate the effectiveness of polyherbal formulation on cataract development in streptozotocin-nicotinamide-induced diabetic rats. Diabetes was induced by streptozotocin (65 mg/kg, ip) and nicotinamide (90 mg/kg, ip). The diabetic rats were orally treated with polyherbal formulation (therapeutic dose, 28.71 mg/kg and 2×therapeutic dose, 57.42 mg/kg) and metformin (150 mg/kg), for 12 weeks. Serum glucose levels, body weight, histopathology of pancreas were used to evaluate the antidiabetic activity while the histopathological study of eye and protein content in lens were used to evaluate the protective effect of polyherbal formulation on cataract development. The progression of cataract in rat lens was observed with a slit-lamp biomicroscope. A daily treatment of polyherbal formulation for 12 weeks resulted in significant (p<0.05) decrease in serum glucose level, body weight and improved protein content in the lens. It also delayed the incidence of cataract in diabetic rats. The histopathological study showed that there was regeneration of pancreatic β-cells. The results observed with polyherbal formulation were found to be comparable to those obtained with metformin. Results suggested that the polyherbal formulation possessed antidiabetic activity and prevented cataract development.

Effect of chronic hyperglycemia on crystallin levels in rat lens

Crystallins are the major structural proteins in the vertebrate eye lens that contribute to lens transparency. Although cataract, including diabetic cataract, is thought to be a result of the accumulation of crystallins with various modifications, the effect of hyperglycemia on status of crystallin levels has not been investigated. This study evaluated the effect of chronic hyperglycemia on crystallin levels in diabetic cataractous rat lens. Diabetes was induced in rats by injecting streptozotocin and maintained on hyperglycemia for a period of 10weeks. At the end, levels of α-, β-, γ-crystallins and phosphoforms of αB-crystallins (αBC) were analyzed by immunoblotting. Further, solubility of crystallins and phosphoforms of αBC was analyzed by detergent soluble assay. Chronic diabetes significantly decreased the protein levels of α-, β- and αA-crystallins (αAC) in both soluble and insoluble fraction of lens. Whereas γ-crystallin levels were decreased and αBC levels were increased in lens soluble fraction with no change in insoluble fraction in diabetic rat lens. Although, diabetes activated the p38MAPK signaling cascade by increasing the p-p38MAPK in lens, the phosphoforms of αBC were decreased in soluble fraction with a concomitant increase in insoluble fraction of diabetic lens when compared to the controls. Moreover, diabetes strongly enhances the degradation of crystallinsand phosphoforms of αBC in lens. Taken together, the decreased levels of crystallins and insolubilization of phosphoforms of αBC under chronic hyperglycemia could be one of the underlying factors in the development of diabetic cataract.

Dynamic and differential regulation in the microRNA expression in the developing and mature cataractous rat lens

Recent evidence supports a role for microRNAs (miRNAs) in regulating gene expression, and alterations in gene expression are known to affect cells involved in the development of ageing disorders. Using developing rat lens epithelial cells (LECs), we profiled the expression of miRNAs by a microarray-based approach. Few gene expression changes known to be involved in pathogenesis or cytoprotection were uniquely influenced by miRNA expression. Most miRNAs increased or decreased in abundance (let 7b, let 7c, miR29a, miR29c, miR126 and miR551b) in LECs/lenses during late embryonic and post-natal development and in cataract. Among them, miR29a, miR29c and miR126 were dramatically decreased in cataractous LECs from Shumiya Cataract Rats (SCRs). Specifically, the cytoskeleton remodelling genes tropomyosin (Tm) 1α and 2β, which have been implicated in the initiation of pathophysiology, were targets of miR29c and were over-stimulated as demonstrated by inhibitor experiments. In transfection experiments, increasing the level of miR29c caused a corresponding decrease in the expression of Tm1α and 2β, suggesting that miR29c may regulate the translation of Tm1α and 2β. 3′UTR luciferase activity of Tm1α, not 2β, was significantly decreased in miR29c-transfected mouse LECs. These findings demonstrate changes in miRNAs expression, and target molecules have potential as diagnostic indicators of ageing and as a foundation of miR-based therapeutics for age-related diseases.

Expression of chloride channel protein-3 and aquaporin-1 in D-galactose cataractous rat lens

目的 研究大鼠半乳糖性白内障晶状体中氯离子通道蛋白-3 (CLC-3)和水通道蛋白-1 (AQP-1) 表达水平的变化,探讨CLC-3和AQP-1与白内障发病的关系.方法 健康清洁级Wistar大鼠88只随机分为空白对照组(8只)、生理盐水组(40只)和半乳糖性白内障模型组(Gal组)(40 只).生理盐水组和Gal组根据处理时间不同各亚分为5个亚组,每个亚组8只大鼠.Gal组单侧眼球后注射D-半乳糖生理盐水注射液,生理盐水组同期单侧眼球后注射等体积无菌生理盐水,每日裂隙灯下观察大鼠晶状体混浊的动态变化,并参照Kador等的分级标准分期记录.分别于第1次给药后第3、7、14、21、30天在全身麻醉下处死各组实验大鼠,摘出晶状体,应用实时荧光定量PCR法检测各组晶状体中CLC-3mRNA和AQP-1mRNA含量的变化.结果 透明晶状体和半乳糖性白内障晶状体中均有CLC-3mRNA和AQP-1mRNA的表达;生理盐水组各时间点CLC-3mRNA和AQP-1mRNA的表达差异无统计学意义(P＞0.05);Gal组于3、7、14d时CLC-3mRNA表达逐渐增高,均高于同期生理盐水组,到14d时约达正常水平的7倍,21d、30d时又有所下降,均低于同期生理盐水组(P＜0.01);各时间点晶状体CLC-3mRNA含量两两比较,差异均有统计学意义 (P＜0.01);模型组于3d时AQP-1mRNA表达已下降,与同期生理盐水组比较差异有统计学意义(P＜0.05),且随着白内障的进一步发展,AQP-1mRNA含量逐渐降低,均低于同期生理盐水组(P＜0.01);各时间点晶状体AQP-1mRNA含量两两比较,差异均有统计学意义 (P＜0.01).结论 晶状体中CLC-3和AQP-1表达的变化可能与白内障的发生发展有着密切的联系。

Gene expression profiling of diabetic and galactosaemic cataractous rat lens by microarray analysis

Osmotic and oxidative stress is associated with the progression and advancement of diabetic cataract. In the present study, we used a cDNA microarray method to analyse gene expression patterns in streptozotocin-induced diabetic rats and galactose-fed cataractous lenses. In addition, we investigated the regulation and interaction(s) of anti-oxidant protein 2 and lens epithelium-derived growth factor in these models. To identify differential gene expression patterns, one group of Sprague-Dawley rats was made diabetic with streptozotocin and a second group was made galactosaemic. Total RNA was extracted from the lenses of both groups and their controls. Labelled cDNA was hybridised to Atlas Rat Arrays. Changes in gene expression level were analysed. Real-time PCR and western analysis were used to validate the microarray results. The expression of 31 genes was significantly modulated in hyperglycaemic lenses compared with galactosaemic lenses. Notably, transcript and protein levels of B-cell leukaemia/lymphoma protein 2 and nuclear factor-kappaB were significantly elevated in rat lenses at 4 weeks after injection of streptozotocin. At a later stage, mRNA and protein levels of TGF-beta were elevated. However, levels of mRNA for IGF-1, lens epithelium-derived growth factor and anti-oxidant protein 2 were diminished in streptozotocin-induced diabetic cataract. These results provide evidence that progression of sugar cataract involves oxidative- and TGF-beta-mediated signalling. These pathways may promote abnormal gene expression in the hyperglycaemic and galactosaemic states and thus may contribute to the symptoms associated with these conditions. Since oxidative stress seems to be a major event in cataract formation, supply of anti-oxidant may postpone the progression of such disorders.

Protection by iodide of lens from selenite-induced cataract.

Iodide has been used empirically against different age-related eye diseases, including cataract. The purpose of the present study was to investigate the effect of iodide on selenite-induced cataract in rat lens. Young white rats received subcutaneously sodium selenite (20 and 30 nmol/g b.w.) on day 13 post partum (p.p.). Cataract development was measured by expert estimation and image data analysis. Potassium iodide (1.5 nmol/g b.w.) was given (1-5 times) i.p. at different times with respect to the selenite administration. Lens opacification was analyzed in selenite, selenite-iodide, iodide and control groups on day 7 after selenite administration. Iodide showed a significant protective effect against selenite cataract when injected 2 days (2 times) before selenite injection, i.e., on days 11 and 12 p.p. No significant effects on lens opacity were found: (1) after only one iodide injection (on day 12 p.p.), (2) after an initial iodide administration 1 h before selenite and (3) after injections of iodide once a day for 5 consecutive days. The protective effect of iodide was the same (about 50%) for both selenite doses used. There is a time-dependent protective influence of iodide against selenite cataract development. It is supposed that the anticataract effect of iodide could be based on direct or indirect antioxidant mechanisms.

Dietary caloric restriction may delay the development of cataract by attenuating the oxidative stress in the lenses of Brown Norway rats

Dietary caloric restriction (CR) is the only experimental intervention that can reliably retard the development of cataract in a normal animal model. Here we have studied the possible mechanisms by which CR retards the age-related degeneration of the lens of Brown Norway rats. We have found that CR slowed protein insolubilization and blunted declines of the total soluble thiols, protein thiols, reduced glutathione and ascorbic acid levels in the lenses of old BN rats. From the lens protein point of view, the development of cataract in rat lenses has 3 stages: (1) the precipitation of γ-crystallin, (2) the insolubilization of β-crystallin, and (3) the final precipitation of α-crystallin which was saturated with other denatured lens proteins. A similar sequence is also observed when the lens proteins are subjected to oxidative stress in vitro. These data are the first to suggest that CR may retard the age-related degeneration of the lens by attenuating the oxidative stress in the lens. Since oxidative stress is likely a main cause of human cataract, CR intervention may be relevant to humans as well.

Vitamin C Metabolomic Mapping in the Lens with 6-Deoxy-6-fluoro-ascorbic Acid and High-Resolution19F-NMR Spectroscopy

Metabolomics, or metabolic profiling, is an emerging discipline geared to providing information on a large number of metabolites, as a complement to genomics and proteomics. In the current study, a fluorine-labeled derivative of ascorbic acid (F-ASA), a major antioxidant- and UV-trapping molecule in the aqueous humor and the lens, was used to investigate the extent to which the lens accumulates potentially toxic degradation products of vitamin C. Human lens epithelial cells (HLE-B3) and rat lenses were exposed to hyperglycemic or oxidative stress in vitro or in vivo and probed for accumulation of F-ASA, fluoro-dehydroascorbate (F-DHA), fluoro-2,3-diketogulonate (F-DKG), and their degradation products in protein-free extracts, by proton-decoupled 750-MHz (19)F-nuclear magnetic resonance (NMR) spectroscopy. F-ASA and F-DHA were taken up into HLE B-3 cells by an Na(+)-dependent transporter. Their uptake was unexpectedly only slightly affected by hyperglycemia in vitro, unless glutathione was severely depleted. Glycemic stress catalyzed oxidation of F-ASA into a single novel F-compound at -212.4 ppm, whereas F-DHA and F-DKG were the major degradation products observed after GSH depletion. In contrast, F-ASA uptake was markedly suppressed in diabetic cataractous rat lenses, which accumulated both the F-DHA and the -212.4-ppm compound. In an unexpected finding, the latter formed only from F-ASA and not F-DHA or F-DKG, suggesting a novel pathway of in vivo F-ASA degradation. Both the cells and the intact rat and human lenses were permeable to several advanced F-ASA and F-DHA degradation products, except F-DKG. The unknown compound at -212.4 ppm was the only F-ASA degradation product that spontaneously formed in rabbit aqueous humor upon incubation with F-ASA. These studies suggest the existence of a novel ascorbic-acid-degradation pathway in the lens and aqueous humor that is influenced by the nature of the oxidant stress. Under similar culture conditions, intact lenses are more prone to hyperglycemia-mediated oxidant stress than are lens epithelial cells, but both are permeable to various F-ASA degradation products, the structure and biological roles of which remain to be established.

Mass Measurements of C-terminally Truncated α-Crystallins from Two-dimensional Gels Identify Lp82 as a Major Endopeptidase in Rat Lens

Molecular chaperone activity of lens alpha-crystallins is reduced by loss of the C terminus. The purpose of this experiment was to 1) determine the cleavage sites produced in vitro by ubiquitous m-calpain and lens-specific Lp82 on alpha-crystallins, 2) identify alpha-crystallin cleavage sites produced in vivo during maturation and cataract formation in rat lens, and 3) estimate the relative activities of Lp82 and m-calpain by appearance of protease-specific cleavage products in vivo. Total soluble protein from young rat lens was incubated with recombinant m-calpain or Lp82 and 2 mM Ca2+. Resulting fragmented alpha-crystallins were separated by two-dimensional gel electrophoresis. Eluted alpha-crystallin spots were analyzed by mass spectrometry. Cleavage sites on insoluble alpha-crystallins were determined similarly in mature rat lens nucleus and in cataractous rat lens nucleus induced by selenite. In vitro proteolysis of alphaA-crystallin by Lp82 and m-calpain produced unique cleavage sites by removing 5 and 11 residues, respectively, from the C terminus. In vivo, the protease-specific truncations removing 5 and 11 residues from alphaA were both found in maturing lens, whereas only the truncation removing 5 residues was found in cataractous lens. Other truncation sites, common to both calpain isoforms, resulted from the removal of 8, 10, 16, 17, and 22 residues from the C terminus of alphaA. Using uniquely truncated alphaA-crystallins as in vivo markers, Lp82 and m-calpain were both found to be active during normal maturation of rat lens, whereas Lp82 seemed especially active during selenite cataract formation. These C-terminal truncations decrease chaperone activity of alpha-crystallins, possibly leading to the observed increases in insoluble proteins during aging and cataract. The methodology that allowed accurate mass measurements of proteins eluted from 2D gels should be useful to examine rapidly other post-translational modifications.

Crystallin proteins in lenses of hereditary cataractous rat, ICR/f.

ICR/f mutation in rat, an inherited disorder, is characterized by the development of cataracts. In this study, we analyzed and compared the crystallins in normal and cataractous rat lenses using gel filtration and two-dimensional gel electrophoresis, and determined the transglutaminase activities and Ca2+ content in the mutant and normal lenses. The Ca2+ content about 10-fold and the activity of transglutaminase was about 1.8-fold higher in the cataractous lenses than in the normal lenses. Analysis of the cataractous lens proteins showed a remarkable decrease in gamma-, betaB1-, betaA3-, and betaA4-crystallin content, accompanied with some increase in alpha-crystallin (or its aggregate). Higher molecular weight proteins were also observed in the cataractous lenses, with molecular masses which correspond to those of cross-linked dimers (43 to 55 kDa) of beta-crystallins. We consider that the mutation accelerates the aggregation of the crystallins, which is associated with their cross-linking by transglutaminase.

Accumulation of 14C-cystine in inherited cataractous rat lens

This study was designed to investigate the formation of mixed disulfides of protein and glutathione (GSH) in the cataractous lens. We compared the changes in accumulation of 14C-cystine in cultured inherited cataractous rat lens (ICR/f) during cataractogensis with those in Wistar strain rats. The accumulation of 14C-cystine in water insoluble protein (WIP) of the lens was increased, especially in lens recognized cataract. The radioactivity accumulated in the WIP was released by incubation with 2-mercaptoethanol (2-ME), dithiothereitol (DTT) and GSH. The accumulation of 14C-cystine in WIP was inhibited by pretreatment with DTT. The existence of some materials in the lens-which combined with S-S compounds became clear. A large part of the materials is present in WIP which is increased along with the lens opacification. We surmised that the accumulation of 14C-cystine was related to the reaction of protein-glutathione disulfide (PSSG).

Lens epithelial cell mRNA. III. Elevated expression of macrophage migration inhibitory factor mRNA in galactose cataracts.

A lens epithelial (LE) cell cDNA clone, designated Clone 156, was isolated from a mature rat LE cDNA library by methods of subtractive hybridization. The cDNA sequence of Clone 156 was 521 nucleotides in length, excluding the poly(T)-tail, and it encoded an open reading frame of 115 amino acids. The translated protein shared extensive sequence similarities with macrophage migration inhibitory factor (MIF) from mouse lens and human T-cell lymphocytes. Northern blot hybridization showed that rat lens MIF mRNA is about 500 nucleotides in length and that its expression in mature rat lens is relatively low in comparison with that in other rat tissues. The expression of MIF mRNA in LE of normal rats and of rats treated by feeding a diet of 50% (w/w) galactose was studied by quantitative RT-PCR. The results showed that the expression of MIF mRNA in a 20-day galactosemic rat LE increased twelvefold as compared to that found in control LE. From the results of this study and from what we know about the locale of epithelial cell differentiation in mature rat lenses, it is being proposed that the increase in abundance of MIF mRNA in the cataractous rat lens is correlated with the enhanced proliferation of the undifferentiated epithelial cells.

Proteolysis by calpain is an underlying mechanism for formation of sugar cataract in rat lens

To confirm the effect of a new aldose reductase inhibitor (ARI), rat lenses were cultured with xylose. ARI prevented opacities and reduced lens hydration caused by xylose. Next, cataract was produced by feeding a diet containing 50% galactose. ARI was tested for amelioration of cataract. On day 19 after feeding of galactose, nuclear cataracts were visible in 75% of the animals receiving only galactose, while nuclear cataracts were not observed in animals treated with ARI. In galactose cataract, lens hydration and calcium were significantly increased. Calpain in soluble and insoluble fractions was decreased. Alpha- and beta-crystallins were proteolyzed. These changes were inhibited by administration of ARI. These results suggested that proteolysis by calpain is an underlying mechanism in formation of sugar cataract in rat lens.

Further studies on the dynamic changes of glutathione and protein-thiol mixed disulfides in H2O2 induced cataract in rat lenses: distributions and effect of aging.

To further investigate the role of protein-thiol mixed disulfides in cataractogenesis, an in vitro H2O2 cataract model was used with rat lenses to study the effect of aging, and the dynamic changes in the cortex, nucleus and the lens protein fractions. A group of lenses was exposed to H2O2-containing media (0.6 mM) for 1 to 3 days so that cortical cataract was induced gradually. Another group of lenses was first subjected to H2O2 exposure for one day and then recovered in the oxidant-free media for one or two days. These lenses were examined for the distribution of free glutathione and protein-thiol mixed disulfides (protein-glutathione and protein-cysteine) in the cortical and nuclear regions as well as in the water soluble and water insoluble fractions. Similar to the results reported earlier, the glutathione depletion in the whole lens occurred immediately and extensively during the 3-day H2O2 exposure. This loss was evenly distributed in the cortical and nuclear fractions. The level of protein-glutathione increased rapidly and continued throughout the 3 days. Most of the accumulation was found in the cortex and in both lens protein fractions. The protein-cysteine modification responded more slowly and less to oxidative stress. The delayed formation occurred mainly in the nucleus and in both lens protein fractions. In the recovery group, glutathione depletion was less drastic in the cortical and nuclear regions, but the elevated protein-glutathione in both regions and both protein fractions spontaneously decreased to its respective basal level within 1 day. Protein-cysteine on the other hand remained quite high, and in some cases it continued to rise in the absence of oxidation. Aging showed little effect on the response of rat lenses to oxidative stress. Similar patterns in glutathione and protein-thiol mixed disulfides occurred in both age groups (1, 23 months) and in both chronic oxidative stress and recovery conditions.

A biochemical study on the effect of Riboflavin on cataractous rat lenses. II Proteins and Glutathione

Proteins and glutathione (GSH) levels were estimated in the experimental groups. Levels of both proteins and glutathione in the control and the treated groups were found to be significantly decreased compared to the normal group. The results show that supplementation of riboflavin has beneficial effect by alleviating the oxidant injuries to GSH and proteins, which are involved in the maintenance of transparency and protection of thiol groups of the lens. Thus, riboflavin has an active role in the lens metabolism and has a potential to improve the metabolic activity of the cataractous lenses.

Abnormal expression of aldose reductase mRNA in fiber cells of cataractous rat lens. Analysis by in situ hybridization.

Aldose reductase (AR), an enzyme of the polyol pathway, has been implicated in the pathogenesis of diabetic and galactosemic cataracts. AR mRNA is a specific transcript of the lens epithelial cells. However, in addition to its presence at high levels in the epithelial cells at the equator, it is also found at significant concentrations in fiber cells at the lens bow. In this study we extended our previous work, and examined the distribution of AR mRNA by in situ hybridization in lens of rats maintained on a 50% galactose diet for up to 20 days, then reversed to a normal diet for an additional 20 days. It was found that within 8 days on galactose the posterior AR mRNA dissipated, with no clear increase in this transcript in the lens epithelial cells. By 16 to 17 days on galactose, the surviving fiber cells in the equatorial region appeared to express AR mRNA; these cells in the normal lens contain insignificant amounts of AR mRNA, as we have previously demonstrated. AR mRNA in the fiber cells of the cataractous lens appears to be of the same base length as that found in the control, indicating that the AR mRNA in these cells may not represent a degraded transcript. Upon reversal of the cataracts, the posterior AR mRNA re-accumulates, and the fiber cells at the equator appear to have lost the AR mRNA that accumulated during the period of exposure to galactose. The data demonstrate that during formation of galactose-cataract, surviving cortical fiber cells express significant levels of AR mRNA, while following withdrawal from galactose these fiber cells lose that activity.

Role of calpain in hydrogen peroxide induced cataract.

The purpose of these experiments was to examine the relationship between oxidation cataract and proteolysis in cultured rat lens. Hydrogen peroxide cataract showed insolubilization of protein, loss of 31 kDa beta B1-crystallin polypeptide, decreases in soluble calpain, and increases in insoluble calpain. This suggested that calpain may be activated in hydrogen peroxide treated lenses, since beta B1 is a known calpain substrate, and calpain undergoes autolysis and degradation when activated. Furthermore, the cysteine protease inhibitor E64 was partially effective in preventing development of H2O2-cataract. E64 also prevented the loss of the 31 kDa beta B1-crystallin polypeptide and decreased the loss of calpain in the lens. These results suggested that development of hydrogen peroxide induced cataract in rat lenses was associated with activation of calpain.

Selective association of crystallins with lens 'native' membrane during dynamic cataractogenesis

Plasma membrane with its associated extrinsic proteins was isolated from normal and cataractous rat lenses by centrifugation of the total water insoluble fraction from homogenized lenses on a discontinuous sucrose gradient. Membrane, which we call "native" membrane, was recovered mainly from the 25/45% sucrose interface. Development of the experimental U18666A cataract resulted in plasma membrane shifting to higher density (the 50/55% sucrose fraction) and great increases in the urea soluble protein content of the lens. At early stages of cataract development, most of the increased urea soluble protein was membrane associated, presumably as extrinsic protein. With advancing cataract, most of the urea soluble protein appeared in an essentially membrane-free pellet fraction. The urea soluble protein associated with the cataract membrane was shown by combined IEF, SDS-PAGE, Western blotting, amino acid compositional analysis and protein sequence determinations to be mainly composed of modified alpha- and beta-crystallins. Alpha A-crystallin truncated by not more than 27 residues from the carboxyl terminus plus beta b1 crystallin truncated by 49 residues from the amino terminus were conclusively identified. In addition to beta b1, a population of six alpha-crystallin derived polypeptides were specifically enriched in the cataract membrane fraction. Four of these six alpha-crystallins appear to be truncated from their carboxyl terminus, a modification which should have increased their hydrophobicity. The pellet fraction, which accumulated in the lens nucleus as the cataract advanced, was enriched in urea soluble gamma-crystallin derived polypeptides. We suggest that protein insolubilization in this experimental cataract involves the selective and tight association of principally modified alpha-crystallins to the fiber cell plasma membrane.

Diagnosis and Estimation of Toxic Changes in the Cataractous Rat Lens induced by Sodium Selenite using Raman Spectroscopy

Diagnosis and Estimation of Toxic Changes in the Cataractous Rat Lens induced by Sodium Selenite using Raman Spectroscopy

Reversal of the limited proteolysis of MP26 during the reversal and prevention of the galactose cataract in rat lenses

The reversal and prevention of the galactose-induced cataract in rats were employed to study their effects on the acceleration of the limited proteolysis of MP26 into MP23-24 previously observed in cataractous lenses of galactose-fed animals. Lenses of rats on a cataract reversal-diet demonstrated the reversal of MP23-24 and MP26 levels to control levels in the clearing cortical areas but not in remaining cataractous nuclear areas. Acceleration of the limited proteolysis of MP26 was observed in the nucleus but not the cortex in the clear lenses of animals on a cataract prevention-diet. The results demonstrated that the limited proteolysis of MP26 may form part of a gradual aging process that although not directly (causally) related to cataractogenesis may at least be accelerated by cataractogenic agents or conditions.