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ROLE OF CASPASE-3 IN ACUTE light damage TO RETINA OF RATS

Chinese Medical Sciences Journal 2007年第1期22卷 44-48页

作者： Xiao Wang Shi-xing Hu Wei Li Shao-chun Lin Zhongshan Ophthalmic Center Sun Yat-sen University Guangzhou 510060 Laboratory Animal Center Guangzhou University of Traditional Medicine Guangzhou 510405

Objective To investigate the role of Caspase-3 in retinal damage caused by hght exposure in rats. Methods light 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 hght 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. Results The 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. Conclusion Apoptosis 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.

关键词： retina light damage Caspase-3 apoptosis rat

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Methanesulfonic acid sodium salt protects retina from acute light damage in mice

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Chinese Medical Journal 2012年第13期125卷 2310-2315页

作者： WANG Hai-yan GU Qing XU Xun Department of Ophthalmology Shanghai First People's HospitalAffiliate of Shanghai Jiao Tong UniversityShanghai 200080China

Background Methanesulfonic 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. Methods One 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 I^m 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. Results The 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

关键词： methanesulfonic acid sodium salt light damage mitochondria apoptosis p 5 3-upregulated modulator of apoptosis

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Retinal damage after exposure to white light emitting diode lights at different intensities in Sprague-Dawley rats

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Annals of Eye Science 2019年第1期4卷 89-96页

作者： Guangsen Wu Xiaoyou Huang He Meng Le Yang Shengjian Lin Yang Gao Yujun Li Yandong Wang Guangzhou No.16 Middle School Guangzhou 510080China Department of Pharmacy&State Key Laboratory of Ophthalmology Zhongshan Ophthalmological CenterSun Yat-sen UniversityGuangzhou 510060China

Background:The usage of the light emitting diode(LED)has been increasingly applied in the illumination setting and electronic ***,the effect of LED lights on the retina remains *** this study,we observed and analyzed the impact of white LED lights at different intensities on the function and morphology of rat ***:Thirty-six Sprague-Dawley rats weighing 150-180 g were randomly divided into six groups(n=6 in each group)including a normal control(NC)group,4 white LED groups at different light intensities(4,000,6,000,7,000,and 10,000 lux),and an ultraviolet B(UVB)lighting group(302 nm,1,000μw/cm2).After 24 hours of continuous illumination,full-field flash electroretinogram(FERG)and pathological examination were performed in each ***:As revealed by FERG,the impairment of retinal function gradually worsened with the increase of LED light *** contrast,the UVB group had the most severe retinal function ***,the functional damage of rod cells and inner nuclear layer cells was the main FERG finding in each *** the NC group,the retina had typical morphologies featured by well-defined structures,clearly visible border between the inner and outer segments,and neatly arranged inner and outer nuclear layer *** 24 hours of illumination,the inner and outer parts of the retina in the 4,000 lux group were still neatly arranged,along with a clear border;however,the inner and outer nuclear layers were randomly arranged,and some irregular nuclei and cells were *** damage of the internal and external retinal segments and the internal and external nuclear layers became more evident in the 6,000 lux group,7,000 lux group,and 10,000 lux *** UVB group had a more obviously disordered arrangement of inner and outer nuclear layers and loss of ***:Continuous exposure to white LED light can cause structural and functional damage to rat retinas,and such damage is related to the intensity of

关键词： Retina light damage light emitting diode(LED) ultraviolet B(UVB) rat

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Effect of EGb761 on light-damaged retinal pigment epithelial cells

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International Journal of Ophthalmology(English edition) 2014年第1期7卷 8-13页

作者： Yun-Yun Zhou Chang-Zheng Chen Yu Su Lu Li Zuo-Hui-Zi Yi Hang Qi Ming Weng Yi-Qiao Xing Department of Ophthalmology Renmin Hospital of Wuhan University Zhongshan Ophthalmic Center Sun Yat-sen University

AIM: To investigate the protective mechanism of Gingko Biloba extract(EGb761) on the ability of retinal pigment epithelial(RPE) cells to resist light-induced damage in a comparative proteomics study. · METHODS: Human RPE cells(ARPE-19) were randomly distributed to one of three groups: normal control(NC group) and light-damaged model without or with EGb761 group(M and ME groups,respectively). The light-damaged model was formed by exposing to white light(2 200 ±300)lx for 6h. The RPE cells in ME group were conducted with EGb761(100μg/mL) before light exposure. The soluble cellular proteins extracting from each groups were separated by two-dimensional electrophoresis and stained by silver staining. Different proteins in the profiles of the gels were analyzed by Image Master Software. Two-fold expressing protein spots were identified by Matrix-assisted laser desorption/ ionization tandem time-of-flight(MALDI-TOF/TOF) mass spectrometry. ·RESULTS: NC,M and ME groups displayed 1 892±71,2 145 ±23 and 2 216 ±85 protein spots,respectively. We identified 33 proteins with different expression levels between the NC and M groups,25 proteins between the M and ME groups,and 11 proteins between the NC and ME groups. MALDI-TOF/TOF mass spectrometry successfully identified 16 proteins,including metabolic enzymes,cytoskeletal proteins,anti-oxidation proteins,and others. ·CONCLUSION: Differences in some important proteins,such as cathepsin B,heat shock protein,and cytochrome C reductase,indicated that multiple pathways may be induced in light-damaged RPE cells and the protective effect of EGb761.

关键词： EGb761 retinal pigment epithelial cells light damage protein mass spectrographic analysis

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EGB761 on retinal light injury in rats

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Chinese Medical Journal 2012年第13期125卷 2306-2309页

作者： QIU Qing-hua XIE Zheng-gao XU Xian LIANG Shu-xian GAO Yan Department of Ophthalmology Shanghai Jiao Tong University Affiliated First People's Hospital Shanghai 200080 China Department of Ophthalmology Northern Jiangsu People's Hospital Affiliated to Yangzhou University Yangzhou Jiangsu 225001 China Shanxi Eye Hospital Taiyuan Shanxi 030002 China

Background Retinal 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. Methods Kunming 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 ***.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. Results In 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） pm. The average thickness of the retina was （126.32~2.31） pm. 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 *** Oral administration of EGB761 can partially inhibit apoptosis of photoreceptor cells, resulting in increased photoreceptor cell survival.

关键词： Ginkgo biloba extract light damage photoreceptor cells retina

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Morphology change of secondary neuron under fast and slow photoreceptor degeneration

Morphology change of secondary neuron under fast and slow ph...

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中国神经科学学会第九届全国学术会议暨第五次会员代表大会

作者： Qiang ZENG~1,Hai-Tao ZHANG~2,Yan YIN~1,Zhi-Ping LIU~1,Yi WANG~2,Zheng-Qin YIN~(2,*),Shi-Gang HE~(1,*) 1 State Key Laboratory of Brain and Cognitive Sciences,Institute of Biophysics,Chinese Academy of Sciences,15 Da- tun Road,Beijing 100101,China 2 Southwest Eye Hospital,Chongqing 400038,China

＜正＞Objective To investigate the influence of photoreceptor degeneration on the morphology of secondary neurons（bipolar and horizontal cells） in the retina of Royal College of Surgeons（RCS） rat and light damaged（LD） albino *** RCS rats and LD mice retinas were sampled at different degeneration stages and compared with wild type（wt） retinas;Both immunohistochemistry and Dil labeling methods were used to examine lamination and morphology of the secondary *** were acquired by confocal microscopy and analyzed with ImageJ. Results Gradual loss of photoreceptors and ribbon synapses was detected from postnatal day 30（P30） to P90 in the RCS rat *** bipolar cells and horizontal cells kept the neurite lamination in outer plexiform layer（OPL） at P60 when 20%photoreceptors left;horizontal cells maintained their morphology at P45 when 50%photoreceptors *** induced fast loss of photoreceptors and ribbon synapses in albino mouse *** and morphology disruption of the bipolar cells happened at 1 day after light damage（DALD）.Morphology change of horizontal cells appeared at 5 *** Structure alternation of secondary neurons in RCS rat retina appears later than photoreceptor loss during the slow degeneration *** in LD mouse retina the alternation appears early following fast photoreceptor loss,indicating the morphology change could happen fast and the residual photoreceptors maintain the structure of secondary neurons.

关键词： retina Royal College of Surgeons light damage horizontal cell bipolar cell

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