OBJECTIVETo study the retinal ultrastructure of streptozocin (STZ)-induced diabetic rats and the intervention effect of Lycium Barbarum Polysaccharides (LBP).

METHODSThe STZ-induced diabetic SD rat model was established. LBP was given to those in the treatment group by gastrogavage. Changes of body weight, blood glucose, and retinal ultrastructure at 24-week were observed.

RESULTSEarly retinal changes covered mitochondrion changes, cell degeneration and apoptosis of retinal neurons and neuroglia cells in the diabetic rats. No change of body weight or blood glucose was observed between the LBP group and the diabetic model group (P > 0.05). The ultrastructural changes were obviously relieved by LBP, and limited to the inner nuclear layer.

CONCLUSIONSLBP could obviously relieve pathological changes of mitochondrion, hinder neural cell apoptosis. Its effect might not be achieved by lowering blood glucose. It was expected to be used in preventing and treating early diabetic retinal neuropathy.

Animals ; Diabetes Mellitus, Experimental ; pathology ; Diabetic Retinopathy ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Retina ; drug effects ; ultrastructure

BACKGROUNDMethanesulfonic acid sodium salt (Dipyrone), an antipyretic and analgesic drug, has been demonstrated to improve cerebral ischemia through the inhibition of mitochondrial cell death cascades. The aim of this study was to evaluate the potential photoprotective activity of methanesulfonic acid sodium salt in a model of light-induced retinopathy.

METHODSOne hundred mice were assigned randomly into vehicle (V), methanesulfonic acid sodium salt (D), light damage model plus vehicle (MV) and light damage model plus methanesulfonic acid sodium salt (MD) groups (n = 25 each). In the MD group, methanesulfonic acid sodium salt (100 mg/kg) was administered by intraperitoneal injection 30 minutes before light exposure. Twenty-four hours after light exposure, hematoxylin and eosin staining and transmission electron microscopy (TEM) were used for histological evaluation. The thickness of the outer plus inner-segment and outer nuclear layer was measured on sections parallel to the vertical meridian of the eye at a distance of 1000 mm from the optic nerve. Electroretinography (ERG) test was performed to assess the functional change. The morphology of mitochondria was also revealed by TEM. Finally, the expression of cytochrome c (CytC) and the relative apoptotic proteins were detected by Western blotting, and the interaction between mitochondrial proteins was investigated by co-immunoprecipitation.

RESULTSThe photoreceptor inner and outer segments of the MV group were significantly disorganized than the MD group. The thicknesses of the outer plus inner-segment layers and the outer nuclear layer, and the amplitudes of the a and b waves of the scotopic ERG response markedly decreased in the MV group compared to those in the MD group (P < 0.05). TEM examination revealed that the mitochondria of the MV group were distinctly swollen and contained disrupted cristae. In contrast, the morphology of mitochondria in the MD group was unaffected. Western blotting analysis showed that CytC, apoptosis proteinase activating factor-1 (Apaf-1), caspase 3, p53, p53-upregulated modulator of apoptosis (PUMA), Bax, and Bad were increased, whereas the anti-apoptotic proteins Bcl-2 and Bcl-X(L) were significantly decreased in the MV group than the MD group. Co-immunoprecipitation detection revealed that PUMA immunoreactivity precipitated by Bcl-X(L) decreased, whereas Bax immunoreactivity precipitated by Bcl-X(L) increased in the MD group compared to those in the MV group.

CONCLUSIONMethanesulfonic acid sodium salt is an effective photoprotective agent against light-induced retinopathy through the inhibition of CytC-mediated mitochondrial impairment.

Animals ; Apoptosis ; drug effects ; radiation effects ; Blotting, Western ; Electroretinography ; Immunoprecipitation ; Light ; adverse effects ; Male ; Mesylates ; therapeutic use ; Mice ; Microscopy, Electron, Transmission ; Mitochondrial Proteins ; metabolism ; Random Allocation ; Retina ; drug effects ; radiation effects ; ultrastructure ; Tumor Suppressor Protein p53 ; metabolism

BACKGROUNDRetinal light injury can lead to degeneration of the photoreceptor cell layer. It has been hypothesized that the mechanism for this process is the photochemical damage. Ginkgo balboa extract (Ginkgo biloba extract EGB761) EGB761 is a free radical scavenger. The purpose of this study was to investigate the possible effect of orally administered EGB761 on retinal light damage of mouse photoreceptor cells.

METHODSKunming mice were randomly chosen for the following groups containing 20 animals in each: control group, light damage group, saline control group, and drug treatment group. The drug treatment group and saline control group were given daily gavage of EGB761 (150 mg×kg(-1)×d(-1)) one week before light exposure. At 7, 14, and 30 days after light exposure, animals were sacrificed and eyes were examined by light microscopy, electron microscopy, and retinal histopathology using in situ detection of apoptotic cells.

RESULTSIn the light damage group after 7 days there was visible edema, and the outer nuclear layer appeared withered with deeply stained dead cells, leaving only a thin nuclear layer of 7 - 8 cells. After 14 days, the photoreceptor cell layer disappeared, leaving only the outer nuclear layer of 1 - 3 cells with an average thickness of (37.988 ± 1.207) µm. The average thickness of the retina was (126.32 ± 2.31) µm. In the drug treatment group, the photoreceptor cell layer and outer nuclear layer damage were significantly lower than the saline group (t = 21.993, P < 0.001), demonstrating that EGB761, especially at 14 days after light exposure, can reduce retinal light damage in mice.

CONCLUSIONOral administration of EGB761 can partially inhibit apoptosis of photoreceptor cells, resulting in increased photoreceptor cell survival.

Animals ; Eye Injuries ; etiology ; prevention & control ; Light ; adverse effects ; Male ; Mice ; Microscopy, Electron ; Photoreceptor Cells ; drug effects ; radiation effects ; ultrastructure ; Plant Extracts ; therapeutic use ; Rats ; Retina ; drug effects ; radiation effects ; ultrastructure

OBJECTIVE: To explore the expression of cyclic guanine monophosphate (cGMP) and the ultrastructure change in retina of guinea pig with form-deprivation myopia and the underlying mechanisms. METHODS: Three-weeks-old guinea pigs were distributed in 3 groups: an untreated group (Group I), a myopia 2-weeks group (Group II) and a myopia 3-weeks group (Group III), animals underwent monocular form-deprivation by facemask for 2 and 3 weeks. The right eyes were deprived and the left eyes were self-controlled. The refraction and axial length of the eyes was measured. Retina was observed by electron microscope. The expression of cGMP was detected by radioimmunochemistry. RESULTS: Deprived eyes in guinea pig showed significant development of myopia, the refraction and axial length was increased. The pathological changes in ultrastructure of retina were aggravated with the development of myopia. The expression of cGMP was significantly up-regulated in the deprived eyes compared with self-control eyes(P<0.05). CONCLUSION Form-deprivation can up-regulate the expression of cyclic GMP, which might play an important role in the development of myopia.
Animals ; Cyclic GMP ; metabolism ; Disease Models, Animal ; Female ; Guinea Pigs ; Male ; Myopia ; etiology ; metabolism ; pathology ; Random Allocation ; Retina ; metabolism ; ultrastructure ; Sensory Deprivation ; Vision, Monocular ; physiology

OBJECTIVETo investigate the injury in the retina of rats exposed to n-hexane.

METHODSThirty-two SD male rats were randomly divided into control group and four n-hexane groups. The rats in the four n-hexane groups inhaled 35.2 g/m3 n-hexane statically for 1, 3, 7 and 14 days respectively (6 rats in every group) while 8 rats in the control group inhaled air. Histopathology and ultrastructure changes of the retina of rats were analyzed.

RESULTSRats in control group had clear layers of retinal structure, stained evenly and with regular cell shape. Retinal degeneration was observed in the rats exposed to n-hexane for 7 d and 14 d, and aggravated by degrees with time exposed to n-hexane. In the rats exposed to n-hexane for 14 d, the outer segments of photoreceptor were arranged in a confusing order, and topically there appeared dissolution; in the inner segments, mitochondria were swollen or disappeared. Pyknotic chromatin and cytoplasmic edema were observed in the outer nuclear layer. There were degeneration of horizontal cells, bipolar cells and amacrine cells in the inner nuclear layer. Cytoplasmic edema and organelle dissolution were observed in ganglionic cells. In the neurofibromas layer, outer and inner plexiform layers, there was neuron cell tuber edema, and the microfilament and vacuole of synapse decreased.

CONCLUSIONThe histopathology and ultrastructure of retina are damaged in the rats exposed to n-hexane, thus leading to ocular fundus disease.

Animals ; Hexanes ; toxicity ; Male ; Rats ; Rats, Sprague-Dawley ; Retina ; drug effects ; pathology ; ultrastructure

OBJECTIVETo investigate the role of oxidative stress in the pathogenesis of retinal injury induced by hyperoxia.

METHODSSixty immature Sprague-Dawley (SD) rats born at a gestational age of 21 days, were randomly exposed to room air (air group, n=30) or 95% oxygen (hyperoxia group, n=30) immediately after birth. Plasma 8-iso-prostaglandin F2alpha (8-iso-PGF2alpha) levels were determined by ELISA. The ultrastructures of the retina were observed under a transmission electron microscope.

RESULTSThe plasma 8-iso-PGF2alpha contents of the air group were 19.09 +/-5.57, 18.24+/-5.91 and 17.00 +/- 5.58 pg/mL on the 3rd, 7th and 14th days after birth, respectively (F=1.024, P> 0.05). The plasma 8-iso-PGF2 contents in the hyperoxia group on the 3rd (28.33 +/- 5.59 pg/mL), the 7th day (51.20 +/- 15.01 pg/mL) and 14th day (84.54 +/- 14.85 pg/mL) after birth were significantly higher than those of the air group (t=2.863, P< 0.05; t=5.073, P< 0.01; t=11.006, P< 0.01). Moreover, the plasma 8-iso-PGF2 contents in the hyperoxia group increased with the prolonged hyperoxia exposure (F=150.7, P < 0.01). The ultrastructures of retina in the air group were normal. Hyperoxia exposure resulted in abnormalities of the ultrastructures of retina, manifesting as the membrane discs rarefied, twisted and disrupted and mitochondrial swelling.

CONCLUSIONSOxidative stress can results in retinal injury in immature rats. An increased plasma level of 8-iso-PGF2alpha is related to the injury degree of retina.

Animals ; Dinoprost ; analogs & derivatives ; blood ; Humans ; Hyperoxia ; complications ; metabolism ; pathology ; Infant, Newborn ; Lipid Peroxidation ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Retina ; metabolism ; pathology ; ultrastructure ; Retinopathy of Prematurity ; etiology

BACKGROUNDGlaucoma is mainly characterized by the loss of retinal ganglion cells. Ciliary neurotrophic factor (CNTF) is believed to stimulate the regeneration of axons of retinal ganglion cells. The objective of our study was to detect the expression of CNTF in the retina of a rat glaucoma model with increased intraocular pressure (IOP).

METHODSThe rat glaucoma model was set up by electrocoagulating at least three episcleral and limbal veins. The location and the expression level of CNTF were detected at 1, 3, 7, 14, and 28 days post-surgery by immunohistochemistry, semiquantitative reverse-transcription polymerase chain reaction (RT-PCR), and Western blot analysis.

RESULTSThe rat glaucoma model with chronic, moderately elevated IOP was successfully produced. A minimum expression of CNTF was found in the ganglion cell layer of the retinas of the control group, and temporally increased expression and intensity of CNTF were found in the experimental retinas.

CONCLUSIONThe expression of endogenous CNTF in the rat retina was found altered after the induction of ocular hypertension.

Animals ; Ciliary Neurotrophic Factor ; analysis ; genetics ; Densitometry ; Immunoblotting ; Immunohistochemistry ; Male ; Ocular Hypertension ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Retina ; metabolism ; ultrastructure ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the role of Caspase-3 in retinal damage caused by light exposure in rats.

METHODSLight injury to retina was induced by persistent exposure to illumination (intensity: 30 000 +/- 50 lux) of operating microscope for 30 minutes in the right eyes of Sprague-Dawley rats. The pathological changes of retina were observed under optical and electron microscopies at different time points, which were 6 hours, 1, 3, 7, and 15 days after the light exposure. Apoptosis of retinal cells was analyzed by flow cytometry. The activity of Caspase-3 was evaluated by using the Caspase-3 assay kit. At the same time, the expression of Caspase-3 protease was determined with Western blot analysis.

RESULTSThe examination results of optical and transmission electron microscopes showed that edema of inner and outer segments of the retina, especially the chondriosome inside the inner segment, became obvious 6 hours after the light exposure. The change was deteriorated along with the increasing time. The structures of the discoidal valve dissociated in the outer segment simultaneously. Disorderly arranged nuclei, karyopycnosis, and thinning in the outer nuclear layer were observed. The retinal pigment epithelium almost disappeared during the later stage. The staining results of Annexin-V combined with PI demonstrated that the proportion of apoptotic cells increased with time. The proportion between 7th day (82.7%) and 15th day (80.4%), however, showed no significant difference. Caspase-3 became remarkably active with the lapse of time, which increased from 0.02 at 6th hour to the peak of 9.8 at 7th day before it started to descend. The Western blot detected a expression of the active form of Caspase-3 at 7th day and 15th day.

CONCLUSIONApoptosis of photoreceptor cells is markedly involved in the light damage and Caspase-3 protease may play an important role in the apoptotic process of the retina after light exposure in rats.

Animals ; Apoptosis ; radiation effects ; Caspase 3 ; genetics ; metabolism ; radiation effects ; Dose-Response Relationship, Radiation ; Enzyme Activation ; Gene Expression Regulation, Enzymologic ; radiation effects ; Light ; adverse effects ; Rats ; Rats, Sprague-Dawley ; Retina ; enzymology ; pathology ; radiation effects ; ultrastructure

OBJECTIVE: To explore the relationship between melanin synthesis and the congenital high myopia of albino guinea-pigs. METHODS: Twelve albino guinea-pigs and 12 pigmented guinea-pigs of 220~250 grams (aged 5~6 weeks) were chosen at random. The eyes were examined with retinoscopy, A-scan ultrasonography and vernier caliper. The retinal structures were examined with light and electronic microscope. RESULTS: The diopter was -19.17 D in albino guinea-pigs and +0.63 D in pigmented guinea-pigs on average. Compared with the pigmented guinea-pigs, the axial dimensions of the albino guinea-pigs were elongated. There was significant difference between the albino guinea-pigs and the pigmented guinea-pigs. The retinal thickness, pigment granules and membrane disc of the outer segment of the visual cells decreased in the albino guinea-pigs, and the membrane disc space became narrow. The normal retinal thickness, plenty of pigment granules , membrane disc and wide membrane disc space could be observed in the pigmented guinea-pigs. CONCLUSION Albino guinea-pigs have high myopia, and pigmented guinea pigs have light hyperopia. There are structural differences in the retina between albino guinea-pigs and pigmented guinea-pigs. The abnormity of albino guinea-pigs provides optical foundation for its high myopia.
Albinism ; complications ; Animals ; Guinea Pigs ; Melanins ; biosynthesis ; Microscopy, Electron, Scanning ; Myopia ; congenital ; etiology ; metabolism ; Random Allocation ; Retina ; metabolism ; pathology ; ultrastructure ; Skin Pigmentation

PURPOSE: To determine the normal range of retinal nerve fiber layer (RNFL) thickness of normal children and adolescents by optical coherence tomography (OCT). METHODS: This study analyzed 144 eyes of 72 normal children and adolescents by OCTIII (Zeiss-Humphrey, San Leandro, CA., USA) and the results were compared with the RNFL thickness of Korean adults. RESULTS: The mean RNFL thickness of the 72 normal children and adolescents was 105.53+/-10.33 micrometer. The mean values for left and right eyes were 104.28+/-7.68 micrometer and 106.79+/-12.98 micrometer, respectively. There was no significant difference in mean RNFL thickness between the 4 quadrants of the left and right eyes (p=0.926). Additionally, the mean RNFL thickness showed a similar size pattern regardless of age (p=0.99). RNFL thickness was found to be greater in adults than in children or adolescents, although the difference was not statistically significant (p=0.295. Likewise, no significant difference was found with gender (p=0.822) or in the pattern of RNFL thickness of 12 sectors between children and adults (p=0.08). CONCLUSIONS: This study reports RNFL thickness, as determined by OCT, for normal children and adolescents. We found this measurement method to be suitable for the early diagnosis of glaucoma and to the examination of its progression in these subjects. The findings could be used as clinical parameters for adolescent glaucoma.
Tomography, Optical Coherence ; Retina/*anatomy & histology ; Reference Values ; Nerve Fibers/*ultrastructure ; Male ; Humans ; Female ; Child ; Adult ; Adolescent