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Expression of P53 during lens epithelial cell Apoptosis Induced by Ultraviolet

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2001年第3期21卷 263-264页

作者：孙旭芳邹尉玉赵长松 Department of Ophthalmology Tongji Hospital Tongji Medical College Huazhong University of Science and Teconology Wuhan 430030

The apoptosis of lens epithelial cells (LECs) induced by ultraviolet and the expression of P53 were investigated. Wistar rats received 100 mW/m 2 ultraviolet irradiation (UVR) (λ=280 nm-315 nm) for 15 min. One, 6, 24 h after irradiation the lens capsules were dissected. The percentages of apoptotic cells were evaluated by the TdT-dUTP terminal nick-end labeling (TUNEL) technique and the expression of P53 was detected by using immunohistochemical assay. The results showed that the percentages of TUNEL-positive nuclei at 24 h after irradiation was significantly higher than in the control group and those 1 h, 6 h after irradiation. The percentages of P53-positive cells at 6 h, 24 h after irradiation were significantly higher than in the control group and those 1 h after irradiation. It was concluded that UVR could induce the apoptosis of lens epithelial cell. The expression of P53 might be responsible for the apoptosis of lens epithelial cells.

关键词： apoptosis lens epithelial cell ultraviolet cataract P53

Overexpression of TRPV1 activates autophagy in human lens epithelial cells under hyperosmotic stress through Ca^(2+)-dependent AMPK/mTOR pathway

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International Journal of Ophthalmology(English edition) 2024年第3期17卷 420-434页

作者： Liu-Hui Huang Jiao Lyu Sheng Chen Ting-Yi Liang Yu-Qing Rao Ping Fei Jing Li Hai-Ying Jin Pei-Quan Zhao Department of Ophthalmology Shanghai Xinhua Hospital Affiliated with Shanghai Jiao Tong University School of MedicineShanghai 200092China Department of Ophthalmology Shanghai Tenth People’s Hospital Affiliated with Shanghai Tongji University School of MedicineShanghai 200072China Centre for Eye Research Australia Royal Victorian Eye and Ear HospitalEast MelbourneVictoria 3002Australia Ophthalmology Department of SurgeryUniversity of MelbourneEast MelbourneVictoria 3002Australia Shenzhen Eye Hospital Shenzhen Key Laboratory of OphthalmologyAffiliated Shenzhen Eye Hospital of Jinan UniversityShenzhen 518040Guangdong ProvinceChina Department of Ophthalmology Shanghai East HospitalSchool of MedicineTongji UniversityShanghai 200120China

●AIM:To explore whether autophagy functions as a cellular adaptation mechanism in lens epithelial cells(LECs)under hyperosmotic stress.●METHODS:LECs were treated with hyperosmotic stress at the concentration of 270,300,400,500,or 600 mOsm for 6,12,18,24h in *** chain reaction(PCR)was employed for the mRNA expression of autophagyrelated genes,while Western blotting detected the targeted protein *** transfection of stub-RFP-sens-GFPLC3 autophagy-related double fluorescence lentivirus was conducted to detect the level of autophagy *** electron microscopy was used to detect the existence of *** interfering RNA of autophagy-related gene(ATG)7,transient receptor potential vanilloid(TRPV)1 overexpression plasmid,related agonists and inhibitors were employed to their influence on autophagy related *** cytometry was employed to test the apoptosis and intracellular Ca^(2+)*** membrane potential was measured by JC-1 *** cell counting kit-8 assay was used to calculate the cellular *** wound healing assay was used to evaluate the wound closure *** 6.0 software was utilized to evaluate the data.●RESULTS:The hyperosmotic stress activated autophagy in a pressure-and time-dependent manner in *** 1 protein expression and conversion of LC3B II to LC3B I increased,whereas sequestosome-1(SQSTM1)protein expression *** Ca^(2+)influx was stimulated caused by hyperosmotic stress,levels of mammalian target of rapamycin(mTOR)phosphorylation decreased,and the level of AMP-activated protein kinase(AMPK)phosphorylation increased in the early *** on this evidence,autophagy activation through the Ca^(2+)-dependent AMPK/mTOR pathway might represent an adaptation process in LECs under hyperosmotic *** stress decreased cellular viability and accelerated apoptosis in LECs and cellular migration *** of autophagy by ATG7 knockdown had sim

关键词： cataract posterior capsular opacification lens epithelial cell hyperosmotic stress autophagy apoptosis transient receptor potential vanilloid 1

Mitochondrial proteomic analysis of ecdysterone protection against oxidative damage in human lens epithelial cells

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International Journal of Ophthalmology(English edition) 2014年第1期7卷 38-43页

作者： Chun-Yan Feng Xiu-Rong Huang Ming-Xin Qi Song-Wen Tang Sheng Chen Yan-Hong Hu Fa-Jie Ke Xin Wang Department of Ophthalmology Second Affiliated Peoples Hospital of Fujian University of Traditional Chinese Medicine Research Center of Pathophysiology Fujian University of Traditional Chinese Medicine Department of Laboratory Fujian University of Traditional Chinese Medicine subsidiary Rehabilitation Hospital

AIM: To investigate the protective effects of the natural medicinal monomer ecdysterone(ECR) with estrogenic activity against oxidative damage in human lens epithelial cells B3(HLE-B3) caused by hydrogen peroxide 21(H2 O2) and to pursue the possible mitochondrial proteomic regularity of the protective effects. · METHODS: HLE-B3 cells were treated with H2O2(300μmol/L),β-estuarial(E2; 10-8mol/L) and H2 O2,ECR(10-6mol/L) and H2 O2,or left untreated. Altered expression of all mitochondrial proteins was analyzed by protein array and surface-enhanced laser desorption ionization time of flight mass spectrometry(SELDI-TOF-MS). The mass/charge(M/Z) ratios of each peak were tested by the Kruskal-Wallis rank sum test,and the protein peak value of the M/Z ratio for each treatment by pair comparison was analyzed with the Nemenyi test. ·RESULTS: H2O2 up-regulated expression of two protein spots(with M/Z of 6 532 and 6 809). When E2 mitigated the oxidative damage,the expression of one protein spot(M/Z 6 532) was down-regulated. In contrast,ECR downregulated both of protein spots(M/Z 6 532 and 6 809). · CONCLUSION: ECR could effectively inhibite H2O2 induced oxidative damage in HLE-B3 cells. The protein spot at M/Z of 6 532 might be the target spot of ECR against oxidative damage induced by H2 O2.

关键词： ecdysterone mitochondrial proteomics lens epithelial cell senile cataract

Surgical stress and cytoskeletal changes in lens epithelial cells following manual and femtosecond laser-assisted capsulotomy

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International Journal of Ophthalmology(English edition) 2020年第6期13卷 927-934页

作者： ANDrea Krisztina Sükosd Krisztina Szabadfi Edina Szabó-Meleg Beáta Gáspár Pavel Stodulka Gyorgy SétálóJr Róbert Gábriel Miklós Nyitrai Zsolt Biró Hajnalka Abrahám Department of Ophthalmology University of Pecs Medical School and Clinical CentrePecs 7623Hungary Department of Experimental Zoology and Neurobiology University of PecsPecs 7624Hungary Department of Biophysics University of Pecs Medical School and Clinical CentrePecs 7624Hungary Janos Szentagothai Research Centre University of PecsPecs 7624Hungary Optimum Laser Centre Budapest 1124Hungary Gemini Eye Clinics Zlin 76001Czech Republic Department of Medical Biology and Central Electron Microscopic Laboratory University of Pecs Medical SchoolPecs 7624Hungary

AIM:To study the effect of mechanical stress on the cytoskeleton in lens epithelial cells following conventional phacoemulsification surgery(CPS)and femtosecond laserassisted cataract surgery(FLACS).METHODS:The cytoskeleton of the epithelial cells of the anterior lens capsules(ALC)removed by CPS and FLACS was examined by *** of the intermediate filament,glial fibrillary acidic protein(GFAP),and glutamine synthetase(GS)immunoreactivity were *** order to map the actin network of cells,fluorescently labeled phalloidin was *** samples were examined using confocal laser scanning ***:GFAP expression was visible in a larger number of the epithelial cells after CPS compared to *** CPS sample’s epithelial cells,GFAP immunoreactivity indicated robust morphological *** the actin filaments,the presence of tubular elements connecting epithelial cells,regular actin pattern and marked cortical network after CPS were *** FLACS,the actin cytoskeleton of the epithelial cells remained densely structured,and the tubular elements were undetectable,however,the above-mentioned regular actin pattern and the marked cortical network were ***:The conventional removal of the ALC induces more robust changes of the cytoskeleton of the lens epithelial cells.

关键词： lens epithelial cell cytoskeleton capsulotomy glial fibrillary acidic protein actin

Expression and proteolytic activity of calpain in lens epithelial cells of oxidative cataract

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Journal of Zhejiang University Science 2004年第6期5卷 743-748页

作者：徐雯姚克孙朝晖王凯军申屠形超 EyeCenter SecondAffiliatedHospitalMedicalCollegeZhejiangUniversityHangzhou310009China

Objective: To study the role of calpain in the mechanism of oxidative cataract through detecting the level of intracellular free Ca2+, the expression and proteolytic activity of calpain in the lens epithelial cells (LECs) of H2O2-induced cataract. Methods: Rat lenses were cultured in vitro and cataract was induced by H2O2. The level of intracellular free Ca2+ was measured by fluorescence determination with fura-2/AM. The expression of m-calpain protein in LECs was detected with immunohistochemical method. The proteolytic activity in LECs was measured using a fluorogenic synthetic substrate. Results: There were significant differences of the level of intracellular free Ca2+ (P=0.001, 0.000, 0.000), the expression of m-calpain (P=0.001, 0.000, 0.000) and the proteolytic activity of calpain (P=0.001, 0.000, 0.000) between H2O2-induced and control group at 6, 12 and 24 h, respectively. Conclusions: H2O2 can increase intracellular free Ca2+, then enhance the expression and proteolytic activity of calpain which may play a role in the mechanism of oxidative cataract of rat.

关键词： Cataract lens epithelial cell Hydrogen peroxide Calpain Calcium

Mitochondrial Proteomic Analysis of Isopsoralen Protection against Oxidative Damage in Human lens epithelial cells

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Chinese Journal of Integrative Medicine 2012年第7期18卷 529-533页

作者：冯春燕黄秀榕祁明信唐宋文胡艳红陈胜柯发杰 Fujian University of Traditional Chinese Medicine Department of Ophthalmology Second Affiliated Peoples Hospital of Fujian University of Traditional Chinese Medicine Research Center of Pathophysiology Fujian University of Traditional Chinese Medicine

Objective： To investigate the protective effects of the natural medicinal monomer isopsoralen （ISR） with estrogenic activity against oxidative damage in human lens epithelial cells B3 （HLE-B3） caused by hydrogen peroxide （H2O2） and to pursue the possible mitochondrial proteomic regularity of the protective effects. Methods： HLE-B3 cells were treated with H2O2 （300 μMol/L）, β-estradiol （E2：10^-8mol/L） and H2O2, ISR （10^-5 mol/L） and H2O2, or left untreated. Altered expressions of all mitochondrial proteins were analyzed by protein array and surface- enhanced laser desorpUon ionization time of flight mass spectrometry （SELDI-TOF-MS）. The mass/charge （m/z） ratios of each peak were tested by the Kruskal-Wallis rank sum test, and the protein peak value of the m/z ratio for each treatment by pair comparison was analyzed with the Nemenyi test. Results： H2O2 up-regulated the expressions of two protein spots （with m/z of 6532 and 6809）. E2 mitigated the oxidative damage, and the expression of one protein spot （m/z 6532） was down-regulated. In contrast, ISR down-regulated both of protein spots （m/z 6532 and 6809）. Conclusions： ISR could effectively inhibit H2O2-induced oxidative damage in HLE-B3 cells. The protein spot at m/z of 6532 might be the target spot of ISR against oxidative damage induced by H2O2.

关键词： isopsoralen mitochondrial proteomics lens epithelial cell senile cataract

Bilateral malignant glaucoma with bullous keratopathy:A case report

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World Journal of Clinical Cases 2023年第13期11卷 3010-3016页

作者： Ya-Bin Ma Ya-Long Dang Department of Ophthalmology Sanmenxia Central HospitalSanmenxia 472000Henan ProvinceChina

BACKGROUND Malignant glaucoma,caused by aqueous misdirection,is a challenging postsurgical complication presented with normal/high intraocular pressure and shallowing of the central and peripheral anterior *** incidence is about 0.6%-4.0%.It can be secondary to filtering surgeries,laser iridotomy,and cataract *** axial length and a history of angle closure glaucoma are its main risk ***,we report a bilateral malignant glaucoma with bullous keratopathy in the patient’s left *** SUMMARY We present a case of bilateral malignant *** cause of malignant glaucoma for each eye of this patient was ***,the management strategy and selection of surgical methods were also ***,the normal anterior chamber was ultimately maintained,and maximum visual function was *** though the left eye received multiple surgeries and corneal endothelial decompensation occurred,the formation of a retroendothelial fibrous membrane partially compensated for the function of the corneal *** The formation of a retroendothelial fibrous membrane partially compensated for the function of the corneal endothelium.

关键词： Malignant glaucoma Corneal epithelial cell lens epithelial cell Bullous keratopathy Retroendothelial fibrous membrane Case report

A Study of Rabbit lens epithelial cells Survival and Growth on the Rabbit Capsular Bag in Vitro

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Journal of Nanjing Medical University 2004年第1期18卷 21-24页

作者：黄瑾谢莉娜卞春及王林农南京医科大学附属南京第一医院眼科南京210006 南京医科大学第一附属医院眼科

Objective: To study the proliferation, migration and metaplasm of residual rabbit lens epithelial cells (LECs) after extracapsular cataract extraction(ECCE)based on the rabbit capsular bag model in vitro. Methods: Sham cataract surgery, including anterior capsulorhexis, nucleus hydroexpression and aspiration of lens fibers, was performed on 20 rabbit lens. The capsular bags were isolated and pinned to sterile non-toxic silicone rings on petri dishes. The capsular bags were incubated with Eagle's minimum essential medium (DMEM) supplemented with 10% fetal calf serum (FCS) and monitored for 3 weeks by phase-contrast microscopy, after which light microscopy was performed on ***: After a latent period of 2-3 d, outgrowth was observed across the posterior capsule. Growth proceeded rapidly so that the posterior capsule was totally covered by a confluent monolayer of cell at 6-8 day. Capsular wrinkles became increasingly apparent as time progressed, causing a marked rise in light scatter. An increase in capsular tension also ***: This model exhibits many of the in vito characteristics of the lens capsule after extracapsular surgery and may prove useful in further elucidating the cellular mechanisms of posterior capsule opacification and developing strategies for inhibiting cell growth with this system.

关键词： lens epithelial cell culture capsular bag model posterior capsule opacification