Inhibition of lens opacification in x-irradiated rats treated with WR-77913

Radiation induced cataracts are models for studying mechanisms of lens opacification. WR-77913, S-3-(amino-2-hydroxypropyl) phosphorothioate (NCS-318809), has been identified as a radioprotective agent. Injection of WR-77913 (1160 mg/kg, i.p.) 15 to 30 min before exposure to 15.3 gray of x-irradiation inhibited rat lenses from developing radiation cataracts. Irradiated rats which did not receive the drug developed dense cataracts. Lenses from control rats which received no radiation remained transparent. Individual lenses were weighed, homogenized, and assayed for protein content using the Lowry method. The molecular weight distribution of soluble protein was determined by HPLC. Mean lens weights were: controls 48.2 mg; irradiated, drug-treated 45.9 mg; and irradiated, nontreated 45.5 mg. Protein accounted for over 40% of the lens weight in control and drug-treated rats and less than 20% for the nontreated cataractous lenses. Water was less than 60% of the lens weight in control and drug-treated rats and over 80% in cataractous lenses. Insoluble protein ranged from 12 to 17% of the total lens weight for each group. The ratio of insoluble to soluble lens protein was 0.40 for control, 0.65 for drug-treated, and 11.28 for cataractous rat lenses. HPLC confirmed a dramatic loss of soluble protein and a complete absence of protein below 25K daltons in cataractous lenses. Proteins below 25K daltons accounted for over 25% of the soluble protein in control and drug-treated rat lenses. WR-77913 stabilizes protein composition and appears to be an effective inhibitor of radiation cataractogenesis.     

Delay of cataract development in hereditary cataract UPL rats by disulfiram and aminoguanidine

The UPL rat is a newly developed hereditary cataract model. We previously found that the administration of disulfiram, a dimer of diethyldithiocarbamate that possesses antioxidant activity, and aminoguanidine, which is known to inhibit inducible nitric oxide synthase, inhibits cataract development in selenite-induced cataract rats. In this study, we investigated the anti-cataract effects and mechanism of disulfiram and aminoguanidine on UPL rats. The opacities of UPL rat lenses, as documented by the anterior eye segment analysis system, EAS-1000 (Nidek, Aichi, Japan), increased from 39 days, and apparently mature cataracts were observed at 53 days. Accompanied with the increase in lens opacity, glutathione concentrations in UPL rat lenses decreased. The Na(+) to K(+) and water-insoluble to water-soluble protein ratios, as well as the Ca(2+) contents in UPL rat lenses increased with the development of cataracts. Oral administration of disulfiram and aminoguanidine delayed the lens opacification as well as the changes in glutathione, Na(+) to K(+) ratio, water-insoluble to soluble protein ratio, and Ca(2+) content in UPL rat lenses. The opacity and Ca(2+) content of UPL rat lenses were closely associated. The present study demonstrates that disulfiram and aminoguanidine have potency of the delay of cataract development in UPL rats, probably caused by inhibiting the rise in Ca(2+) levels.     

Quantitative estimation of the relationship between amino acids and cataracts in rat lens]

The concentration of free amino acids and their related compounds has been determined in the lenses of ICR (f) strain rat and in the Wistar strain rat's lenses which were cultured with diethyl maleate. It was supposed that the decrease of cystathionine and the increase of serine in lenses of ICR with aging were related with development of senile cataracts. The increase of cystathionine in lenses cultured were suggested that synthesis of taurine is done by cystathionine pathway. Quantitative changes of amino acids were higher than normal of glutamine, glycine and aspartate in lenses cultured. It was supposed that the changes were the flow in lens from medium for synthesis of glutathione and glucose.     

Adsorption and removal of protein bound to hydrogel contact lenses

Tear film proteins are known to adsorb to new hydrogel contact lenses. Using a radioiodine tracing technique, proteins were shown to adsorb to contact lens surfaces. The quantity of protein adsorbed to the contact lenses within 2 to 4 h was 1 to 3 micrograms/lens. The degree of protein adsorption varied from lens-to-lens depending in part on the water content of the lens. High water content ionic lenses bound the most protein, whereas lower water content and nonionic lenses bound less protein. Enzyme cleaning of the protein-coated contact lenses removed about 75% of the adsorbed protein. When the enzyme-cleaned lenses were reincubated with protein, similar quantities were quickly readsorbed within a similar period of time. The rate of readsorption appeared to vary with the type of enzyme used to clean the lenses. Hydrogel lenses adsorb protein from the tear film rapidly and irreversibly. Cleaning the lenses has only a partial and temporary effect on the bound protein. The lens-bound proteins could become partly denatured during their binding to the lenses. These observations suggest a possible role for such proteins in ocular sensitivity to contact lenses.     

Lens growth and protein density in the rat lens after in vivo exposure to ultraviolet radiation

PURPOSE: To investigate lens growth after different doses of ultraviolet radiation (UVR) and to investigate the long-term effect of a near-threshold UVR dose on the refractive index distribution in the lens. METHODS: Sprague-Dawley rats received UVR (lambda(MAX) = 300 nm) unilaterally during a 15-minute period. The exposure dose ranged from 0.1 to 20 kJ/m(2), and the rats were kept for up to 32 weeks after exposure. Intact lenses were photographed and lens wet and dry masses were measured. The protein density was estimated by quantitative microradiography. Freeze-dried lens sections were used for contact x-ray photographs. From the transmission of the microradiographs, protein density and refractive index profiles were calculated along the lens radius with a resolution of 2.5 microm. RESULTS: Lens dry mass in exposed eyes was lower than in nonexposed eyes at one week after exposure. Lens water content was decreased after low UVR doses but increased after high doses. The difference between exposed and nonexposed lenses in dry mass and water content increased with time after exposure. No significant difference was found for the mean protein density in exposed and nonexposed lenses. The protein density increased linearly in the lens cortex, from a minimum in the superficial cortex of 0.26 g/cm(3) to a maximum in the deep cortex of 0.81 g/cm(3). This corresponded to a refractive index of 1.38 and 1.48, respectively. CONCLUSIONS: Lenses exposed to UVR grew more slowly than their nonexposed contralaterals. This growth inhibition was dose dependent. Near-threshold doses led to decreased water content in the lens whereas high doses led to swelling. Six months after near-threshold UVR exposure, no global change of the refractive index was found. However, local variations of the refractive index caused a subtle cortical light scattering.     

Protein oxidation and lens opacity in humans

PURPOSE: Oxidative damage to lens proteins is a major factor leading to cataract formation. It is of pathogenic importance to determine a threshold of protein oxidation over which opacification of the lens takes place. METHODS: Sixty-two lenses extracted from patients affected by idiopathic senile, diabetic, or myopic cataract were studied. Clear lenses were obtained from subjects undergoing enucleation (n = 10) or vitrectomy for giant retinal tears (n = 9), and were age- and sex-matched to those with cataract. The content of carbonyls and sulfhydryls (P-SH) in proteins in the lens was assessed using spectrophotometric assay. RESULTS: An age-associated inverse relation (P < 0.01) was noted in the content of P-SH, the concentrations of which were also inversely related (P < 0.03) to the content of protein carbonyls. These changes were more pronounced in cataracts than in clear lenses and in diabetic and myopic cataracts when compared with senile cataracts. The drop of P-SH concentration occurred earlier in diabetic and in myopic cataracts than in senile cataracts. The accumulation of protein carbonyls > 2 nmol/mg protein and the decrease of P-SH below 12 to 10 nmol/mg protein were always accompanied by lens opacification. CONCLUSIONS: Idiopathic senile, diabetic, and myopic cataractogenesis appear to be dependent on oxidative damage to lens proteins. This damage occurs earlier in myopic and diabetic patients. Values of P-SH below and protein carbonyls above their specific threshold were found to be predictive for the presence of cataract. Because increased oxidation was observed in clear lenses removed from myopic and diabetic subjects, oxidation may be involved in the pathogenesis of these forms of human cataract.     

人類不同年齡晶體巰基含量的變化和年齡相關性白内障形成的關系

目的使用傳統的巰基顯色劑Ellman試劑測量并比較分析人類不同年齡段的透明及各類型白内障晶狀體樣本中非蛋白巰基(游離巰基)、蛋白巰基和蛋白結合巰基的含量,并分析其與人類年齡相關性白内障形成的關系.方法44個人類晶體按年齡大小被分爲5組,其中10個白内障晶體被分成2組(皮質性和核性).用Ellman試劑測量以上樣品的晶體非蛋白巰基、總蛋白巰基和總蛋白結合巰基含量.結果 皮質性白内障晶狀體的非蛋白巰基含量顯著高于核性白内障晶狀體(P＜0.01);而在蛋白巰基和蛋白結合巰基中,兩者則無顯著差别(P＞0.05).非蛋白巰基含量從胚胎期到年齡相關性白内障發生逐漸降低,各年齡組均顯著小于前一年齡組(P＜0.01);蛋白巰基含量在第5組顯著高于第1、2、3、4組(P＜0.05),其余各組間無顯著區别(P＞0.05);蛋白結合巰基含量除第3、4組和第4、5組間無顯著區别外(P＞0.05),其余組均顯著低于較小年齡組(P＜0.01).結論 人類年齡相關性白内障的形成過程中,晶狀體蛋白分子間巰基的氧化并不像在其它實驗性白内障模型中那樣起决定作用.     

Determination of the state and content of water in normal avian, fish, porcine, bovine, and human lenses as studied by differential scanning calorimetry

Differential scanning calorimetry was used to measure the relative amounts of 'bulk' and 'bound' water in normal avian, bovine, fish, human, and porcine lenses. The amounts of bound water (mg bound water/mg lens dry weight) found in avian and porcine lenses were statistically different from each other in addition to being statistically different from fish, human, and bovine lenses. There were no significant differences in the mean values between human, fish, and bovine lenses. Avian lenses had the highest amount of bound water, while fish lenses had the lowest bound water content. Significant differences in total water content (mg total water/mg lens dry weight) were observed between all of the lenses, with the exception of bovine and human lenses which were not statistically different. Fish lenses had the lowest amount of total water, and avian lenses had the highest total water content. There were significant differences in bulk water content (mg bulk water/mg lens dry weight) between all of the lenses. Avian lenses had the highest bulk water content, and fish lenses had the lowest bulk water content.     

Methionine adenosyltransferase and S-adenosylmethionine in the developing rat lens

Methionine adenosyltransferase (MAT) activity, and the concentration of its reaction product, S-adenosylmethionine (AdoMet), were measured in the lenses of rats of different ages; ranging from 1 day of age to over 1 yr. The highest specific activity of MAT was found in the lenses of the one day old rats (sp. ac. 0.327 units/mg-1 protein). After 1 week the specific activity had dropped to 0.067, and by 6 weeks had declined to adult levels (0.02 units/mg-1 protein). AdoMet concentrations were measured by HPLC in perchloric acid extracts. The highest concentration of AdoMet was found in the lenses of day-old rats (48.2 microM), and gradually declined with increasing age, reaching 5.5 microM in the oldest rats. In addition, the specific activity of MAT was found to be higher in the lens epithelium than in the cortex plus nucleus. The specific activity of MAT is almost an order of magnitude higher in the lens epithelial fraction (0.099 units mg-1 protein) than in the combined cortex plus nucleus fraction (0.011 units mg-1 protein).     

Protein kinase C-gamma activation in the early streptozotocin diabetic rat lens

PURPOSE: The purpose of this study is to demonstrate the early activation of the protein kinase C-gamma (PKC-gamma) pathway in the streptozotocin (STZ)-induced diabetic rat lens. METHODS: Twelve-week-old male and female Sprague-Dawley rats were injected with 80 mg/kg (body weight) of STZ (N-[methylnitrosocarbamoyl]-D-glucosamine) intraperitoneally. Very high glucose (VHG) diabetes was defined as a nonfasting blood glucose level of at least 450 mg/dl, confirmed by daily monitoring with Accu-Check Advantage test strips, and occurred about 2 weeks after STZ administration. All assayed lenses were from VHG or age-matched control rats, harvested within 24 hr of VHG detection. PKC-gamma activation was measured by enzyme activity assay and by Western blotting to show autophosphorylation on Thr514. Cellular insulin-like growth factor-1 (IGF-1), PKC-gamma phosphorylation of Cx43 on Ser368, and activation of phospholipase C-gamma 1 (PLC-gamma 1), extracellular signal-regulated kinase (ERK1/2), and caspase-3 were determined by Western blotting. Endogenous diacylglycerol (DAG) levels were measured with a DAG assay kit. Lens gap junction activity was determined by the microinjection/Lucifer yellow dye transfer assay. Electron microscopy was applied to affirm fiber cell damage in the VHG diabetic lenses. RESULTS: In the lenses of VHG diabetic rats, PKC-gamma enzyme was activated. PKC-gamma could be further activated by 400 nM phorbol-12-myristate-13-acetate (PMA), but the PKC-gamma protein levels remained constant. No elevation of IGF-1 level was observed. Western blots showed that activation of PKC-gamma may be due to activation of PLC-gamma 1, which synthesized endogenous DAG, a native PKC activator. The level of PKC-gamma -catalyzed phosphorylation of Cx43 on Ser368 and resulting inhibition of lens gap junction dye transfer activity was increased in the VHG diabetic lenses. At this early time period, the diabetic lens showed no activation of either caspase-3 or ERK1/2. Only a single fiber cell layer deep within the cortex (approximately 90 cell layers from capsule surface) showed vacuoles and damaged cell connections. CONCLUSIONS: Early activation of PLC-gamma 1 and elevated DAG were observed within VHG diabetic lenses. These were correlated with activation of PKC-gamma, phosphorylation of Cx43 on Ser368, and inhibition of dye transfer. Abnormal signaling from PKC-gamma to Cx43 in the epithelial cells/early fiber cells, observed within VHG diabetic lenses, may be responsible for fiber cell damage deeper in the lens cortex.     

Lens proteins in intumescent cataract

Intumescent cataractous lenses were investigated as to their water content and protein composition. Nuclei were separated from cortices. Water-soluble, water-insoluble, urea-soluble and urea-insoluble portions were quantified and the subunit composition of the water-soluble crystallin fraction was looked at on SDS-gel electrophoresis. Compared to normal lenses it is observed that the water content in intumescent cataract is increased, water-soluble and urea-soluble fractions are diminished, whereas the urea-insoluble portions are augmented. No major changes are noted in the subunit composition of the soluble protein fractions. Particularly, the relative amount of gamma-crystallins is diminished in intumescent cataractous lenses.     

Partial reversal of methylprednisolone-induced opacity in isolated rat lenses

We examined whether opacity is reversed in isolated rat lenses with opacities of different grades induced by methylprednisolone treatment by withdrawal of the steroid treatment. Transparent lenses isolated from male Wistar rats aged 6 weeks were incubated in TC-199 medium containing methylprednisolone hemisuccinate (1.5 mg/ml) for 24 or 48 h to induce opacities of two different grades (one was a slight cortical opacity around the equator and the other a heavy cortical opacity spreading from the equator to the cortex). In the slightly opaque lenses, water content was preserved at the level of transparent lenses untreated with steroid, although a 3.2-fold increase in Na(+)/K(+) ratio and a 20% decrease in Na(+),K(+)-ATPase activity were observed. In contrast, in the heavily opaque lenses, a significant increase in water content was accompanied by a 31.0-fold increase in Na(+)/K(+) ratio and a 27% decrease in Na(+),K(+)-ATPase activity. When the slightly and heavily opaque lenses were further incubated in steroid-free medium for 24 or 48 h, a partial opacity reversal was found in the slightly opaque lenses, but not in the heavily opaque lenses. In the slightly opaque lenses incubated in steroid-free medium for 48 h, increased Na(+)/K(+) ratio and decreased Na(+),K(+)-ATPase activity were restored to nearly the levels of untreated transparent lenses. Incubation of the heavily opaque lenses in steroid-free medium for 48 h caused further slight increases in water content and Na(+)/K(+) ratio and further slight decrease in Na(+),K(+)-ATPase activity. These results indicate that opacity is partially reversed in rat lenses with a slight opacity induced by methylprednisolone hemisuccinate when the steroid treatment is withdrawn, and suggest that recovery of impaired lens membrane function could be involved in this in vitro partial opacity reversal.     

体外培养大鼠白内障模型晶状体的早期生化改变

目的 进一步探讨白内障的发生机制。方法 采用器官培养方法,以不同浓度亚硒酸钠及半乳糖作用于晶状体,诱发大鼠晶状体形成白是,在培养初期测定晶状体内非蛋白质疏基（ｎｏｎｐｒｏｔｅｉｎｓｕｌｆｈｙｄｒｙｌ,ＮＰ－ＳＨ）、蛋白质疏基（ｐｒｏｔｅｉｎ ｓｕｌｆｈｙｄｒｙｌ,Ｐ－ＳＨ）和不溶性蛋白质二硫键的含量,脂类过氧化水平,以及与谷胱甘肽代谢有关的谷胱甘肽过氧化物酶（ＧＳＨ ｐｅｒｏｘｉｄａｓｅ,ＧＳＨ－     

Age dependent lipid and protein changes in individual bovine lenses

Successive fiber fractions isolated from individual bovine lenses were fractionated to examine changes in lens proteins and membrane lipids as a function of age. In calf lens of about 1.2 gm wet weight, cholesterol (C) is maintained at a level of 3.3 microgram/mg dry weight in the outer cortical 30% of the lens. In the inner cortex, a C content of 2.4 micrograms/mg was found that decreased somewhat to 2.1 micrograms/mg in the inner nuclear 20% of the lens. The almost linear decrease in phospholipid (P) content from 11.6 in the cortex to 1.7 micrograms/mg in the nucleus resulted in a cortex to nucleus increase in C/P ratio from 0.5 to about 2.0 (mol/mol). Compared to calf lenses, a low C level of 2.4 micrograms/mg was observed in the outer cortex of cow lenses (approximately 3.0 gm wet weight). No significant difference in C level was found between the calf and cow lenses either in the inner cortical or nuclear regions. The P level was reduced to 6 and 1.2 micrograms/mg in the outer cortex and nucleus of the cow lens, respectively. The low nuclear P content is responsible for the observed high C/P value of 3.6. The lower lipid content found in the cortex of older lens suggests an age dependent decrease in the amount of available membrane lipid to envelope the newly formed fibers. A cortex to nucleus increase in the amount of urea-soluble (US) protein fraction from about 6 to 14% of total fiber mass was observed with the calf lens. In the cow lenses, the nuclear US fraction accounts for almost 30% of the fiber.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relative importance of aldose reductase versus nonenzymatic glycosylation on sugar cataract formation in diabetic rats.

The relative importance of sorbitol formation versus nonenzymatic glycosylation and advanced glycosylation end products (AGEs) on sugar cataract formation was examined in diabetic rats. Diabetes was experimentally induced in young, 50 g rats with streptozotocin, and aldose reductase inhibitors were administered in the diet for up to 8 weeks at concentrations of 0.06% for tolrestat or ponalrestat and 0.0125% for AL-1576. Cataract formation was monitored by hand-held slit lamp for up to 11 weeks. Lens polyol levels were monitored by GLC, glycosylated protein levels were spectrophotometrically determined, and AGE products were estimated by fluorescence measurements and ELISA. Sugar cataract formation was observed in all untreated diabetic rats while cataract formation was inhibited in all diabetic rats treated with the AR inhibitors. Lens sorbitol levels were reduced in all ARI-treated rats. Glycosylated lens protein levels were elevated in the diabetic rats, and these levels were not significantly lower in the non-cataractous lenses from ARI-treated diabetic rats. Fluorescence measurements of the lens proteins revealed increased lens AGE levels in all diabetic rats, and these were slightly reduced in the aldose reductase inhibitor treated diabetics. With ELISA, immunoreactive AGEs were only detected in cataractous lenses from the untreated diabetic rats. Immunoreactive AGEs were not detected in the clear lenses of the aldose reductase inhibitor treated diabetics or in the non-diabetic controls. These results support the concept that sugar cataract formation is initiated by the aldose reductase catalyzed intracellular accumulation of polyols and that these sugar cataracts can be prevented through inhibition of aldose reductase.     

Aminopeptidase III activity in normal and cataractous lenses

Aminopeptidase III activity was demonstrated in extracts from several different mammalian lenses by the hydrolysis of Arg-MCA at pH 6.0. No more than a two-fold difference was seen in overall specific activity. Sections of bovine lenses were removed from the periphery to the center and assayed. A sharp decline in activity was observed in the inner cortical region, and little or no activity was observed in the lens nucleus. This correlated with an increase in the presence of low molecular weight peptides as determined by SDS polyacrylamide gel electrophoresis. The properties of the aminopeptidase from human lens tissue were the same as those previously reported for the purified enzyme from bovine lens. The aminopeptidase activity of normal and cataractous lenses was compared using 4 different substrates. The cataractous lenses had significantly less total aminopeptidase activity. However, little difference in specific activity was observed based on soluble lens protein content. Similarly, electrophoretic separations of normal and cataractous soluble proteins showed little or no differences in the content of low molecular weight peptides. Therefore, this major human lens aminopeptidase remains functional in the cataractous state.     

Distribution of total and non-freezable water contents of galactosemic rat lenses

Galactosemic cataracts were induced in Sprague-Dawley rats by keeping them on a 50% galactose diet for 36 days. The galactosemic rat lenses as well as normal rat lenses were sliced into 0.50 mm sections and differential scanning calorimetric (DSC) and thermogravimetric (TGA) measurements were performed. The TGA measurements gave the total water content. This was much higher in cataractous lenses than in normal lenses. The total water content also showed different gradients in normal and cataractous lenses. In normal lenses the percentage of water decreases as one proceeds from cortex to nucleus. In galactosemic cataractous lenses the trend is the opposite. The non-freezable (bound) water content was obtained as the difference between the total and freezable (DSC measurements) water content. In both cataractous and normal lenses the non-freezable water content increases when one proceeds from cortex to nucleus. It seems that most of the water influx generated by the accumulation of galactitol ends up as free water in lakes and pools rather than water of hydration.     

Biochemical evidence for conversion to milder form of hereditary mouse cataract by different genetic background

Congenic hereditary cataract mice, BALB/c-nct/nct, were established by introducing the nct gene from Nakano into BALB/c mice. These mice developed a milder cortical form of cataract which developed sporadically and later in life than in Nakano mice. Combined use of BALB/c and BALB/c-nct/nct mice enables biochemical comparison of normal clear lenses, congenic clear lenses which are destined to be opacified some time later, and opacified lenses in the same genetic and aging statuses. We compared the age-related changes in water content and water-soluble and -insoluble fractions among these three types of lenses. Congenic clear lenses and opaque lenses were more similar to BALB/c normal clear lenses and Nakano opaque ones, respectively, in these parameters. These results suggest, in addition to formation of aggregated crystallins and their accumulation in water-insoluble fractions, that decreased protein synthesis, increased protein degradation and augmented leakage of crystallin might have a significant role in the nct-induced lens opacification.     

Deferoxamine effect on selenite-induced cataract formation in rats.

A single subcutaneous dose of 30 nmol of sodium selenite per gram of body weight in 13-day-old rats resulted in posterior subcapsular cataract (PSC) after 24 hr and bilateral nuclear cataracts at 72-96 hr. Within 24 hr of treatment, a 60% decrease in lens glutathione was seen. A loss of calcium homeostasis observed by 48 hr resulted in increased lens calcium (4 mumol/g dry weight), which accompanied nuclear opacification. The iron chelator, deferoxamine (DF), was evaluated as a potential protective agent against these selenite-induced changes. Three doses each consisting of 1.1 mumol DF/g body weight were administered during the initial 24 hr of selenite exposure. Within 96 hr, all lenses from animals treated only with DF remained transparent, but 50% of these lenses showed cortical cataract at 3 wk postinjection. Concurrent administration of DF and selenite protected 80% of rats against PSC after 48 hr and 25% against nuclear cataract after 96 hr. No elevation in lens calcium occurred in the protected lenses. An additional 20% of animals were not protected fully but showed substantially less nuclear opacity than with selenite alone. They had a significant but moderate increase in lens calcium. After 3 wk (animal age, 35-40 d), cataract appeared in these "protected" lenses involving both the nucleus and cortex and loss of ion homeostasis. The glutathione content remained lower in lenses from animals treated with both selenite and DF compared with those from selenite-treated animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

波形纤维蛋白在其转基因小鼠晶状体的表达

目的 观察波形纤维蛋白（Ｖｉｍｅｎｔｉｎ）在转基因小鼠晶状体的表达,探讨波形纤维蛋白与内障形成的关系。方法 用显微注射法将鸡的１２．７ｋｂ波形纤维蛋白基因导入小鼠受精卵的雄前核内,经培育、杂交传代得到２０只波形纤维蛋白内障小鼠。取４只白内障眼球及４只正常眼球,制备石蜡切片,采用ＡＢＣ免疫组化方法,观察波形纤维蛋白在晶状体的表达。再分别正常、分部混浊和完全混浊的小鼠晶状体各１只,经ＳＤＳ－聚丙烯酰胺