Effect of biological amniotic membrane soaked in pirfenidone on scar formation after glaucoma filtration surgery in rabbit eyes / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the application of biological amniotic membrane soaked in pirfenidone(PFD)and to evaluate its anti-scarring effect and toxic side effects on glaucoma model of rabbit eyes.METHODS: The right eyes of 72 healthy New Zealand white rabbits were randomly divided into 0.5%PFD+ biological amniotic membrane group, biological amniotic membrane group, mitomycin C(MMC)group and blank control group after the glaucoma model was established by anterior chamber injection of compound carbomer solution, and 18 rabbits in each group underwent trabeculectomy, in which the 0.5% PFD+ biological amniotic membrane group was placed with 0.5% PFD solution-soaked biological amniotic membrane under the scleral flap, and the biological amniotic membrane group was placed with normal saline-soaked rehydrated biological amniotic membrane under the scleral flap. In the MMC group, a cotton pad soaked in MMC was placed under the scleral flap for 3 min and immediately rinsed with normal saline, while the blank control group received no implant after the scleral flap was made. The intraocular pressure(IOP), filtration blebs, toxic side effects and complications were evaluated, and the histopathological changes in the filtration area were observed by hematoxylin-eosin(HE), Masson staining and immunohistochemical staining.RESULTS: The mean IOP at 14, 21 and 28 d after trabeculectomy were 0.5%PFD+ biological amniotic membrane group<MMC group<biological amniotic membrane group<blank control group(all P<0.05). At 28 d after trabeculectomy, 0.5%PFD+ biological amniotic membrane group had the best effect of anti-inflammatory hyperplasia and inhibition of collagen formation, the highest survival rate of filtration blebs, and the inflammatory reaction was mild.CONCLUSION: Biological amniotic membrane soaked in pirfenidone has more obvious anti-scarring effect on glaucoma model, with less toxic side effects and good safety.

Effect of RSCs combined with COP-1 on optic nerve damage in glaucoma rat model

OBJECTIVE@#To explore effect of retinal stem cells (RSCs) combined with copolymer-1 (COP-1) immunotherapy on optic nerve damage in glaucoma rat model.@*METHODS@#A total of 40 SD rats were selected for glaucoma model and were randomly divided into 4 groups to observe protective effects of RSCs transplantation combined with COP-1.@*RESULTS@#Brain-derived neurotrophic factor (BDNF) and insulin like growth factor-1 (IGF-1) were either positive in retina of RSCs transplanted or COP-1 immunological treated rat. Positive rate of BDNF and IGF-1 and expression of mRNA and protein were significantly higher in RSCs transplantation combined with COP-1 immunotherapy treated rats compared with the other 3 groups, in which amount of apoptotic RGCs was lowest.@*CONCLUSIONS@#RSCs transplantation combined with COP-1 immunotherapy can promote the secretion of BDNF and IGF-1. They protect RGCs in glaucoma rats in coordination, significantly reduce the number of apoptosis RGCs so as to alleviate the optic nerve damage. It ponits a new research direction for treatment of glaucoma.

The dynamic change of Brn-3a expression in retina of chronic high intraocular pressure rat / 中华实验眼科杂志

Background Brn-3a is a newly discovered specificity marker for retina ganglion cells(RGCs).It is well-known that RGCs damage is a important pathological basis of hypertension-visual disorder.But the study concerning expression of Brn-3a in RGCs in glaucoma eye is still rately.Objective The purpose of this work was to investigate the changes of Brn-3a expression in model eye with chronic high intraocular pressure(IOP) and its relation with morphology of retina and the expression of Brn-3a in chronic ocular hypertension rats.Methods Thirty-five clean adult SD rats were randomly divided into normal control group(5 rats) and model group(30 rats).Experimental chronic ocular hypertension models were induced unilaterally in the left eyes of 30 health adult SD rats by cauterizing super-scleral veins,and the conjunctival incision was made in the right eyes as sham operative group.The operated rats were subdivided into 6 groups according to the examination time points and 6 rats for each group.IOP was measured with Tono-Pen tonometer before and after 30 minutes,1,3,7,14,28 day after surgery respectively.The rat models were sacrificed in 1,3,5,7,14,28 days after operation by excessive anesthesia method,and retinal section was prepared for the histopathological examination and the RGCs were counted using Nissl staining method.Expression of Brn-3a in RGCs was detected by immunohistochemistry.This experimental complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The IOP was significantly raised from 30 minutes to 28 days after operation in model eyes with the top IOP( 34.46±4.65 )mmHg in the 30 minutes after operation,showing statistically significant differences in various time groups ( F =95.631,P =0.001 ) and different eyes ( F =287.473,P =0.001 ).Compared with sham operative group,the IOP were elevated from 1 day through 14 days after operation ( q =18.418,15.261,10.987,6.931,4.975,2.962,P ＜ 0.05 ).The numbers of RGCs were ( 29.08 ± 1.98 ) in the normal control group and decreased gradually by 3.17％,7.84％,14.60％,22.40％ in 1,7,14,28 day after surgery in the model eyes with the considerable differences in comparison with normal eyes ( t =5.943,8.034,15.023,17.004,19.371,P ＜ 0.05 ).Immunochemistry revealed that Brn-3a was specially expressed in the RGCs layer and the positive RGCs for Brn-3a were evidently decreased as the prolong of high IOP duration ( F =127.583,P =0.000 ).Conclusions Chronic glaucoma model can be successfully established using Shareef-Sharma method.The loss of RGCs is associated with the high lOP duration.The expression level of Brn-3a is a reliable index of high-IOP-induced damage of optical nurve.

Ocular findings and histological change in glaucoma model of DBA/2J Mice / 眼科研究

Background As a hereditary chronic glaucoma model,DBA/2J mouse is widely used in the experimental research of glaucoma worldwide.Although some researches have determined the ocular change induced by hypertention of DBA/2J mouse,there is seldom reports about the research on the relationship of ocular pathological abnormality and development course in DBA/2J mouse.Objective The present study is to characterize the ocular abnormalities and histological changes induced by hypertention in different ages of DBA/2J mice.Methods The clean female DBA/2J mice aged 3-,5-,7-,9-,11-,14-month-old (6 mice for each) were used in this study,and age matched 18 female C57BL/6J mice were as controls.Intraocular pressure (IOP) of mice was measured by anterior chamber puncture of microneedle.The animals were sacrificed and retinal flat mounts were prepared for histopathological examination under the light microscope.Retinal ganglion cells (RGCs) were counted by retinal Nissl staining.Morphology of optic nerve cup in frozen section was examined under the light microscope.The experiment followed the Standard of Association for Research in Vision and Ophthalmology.Results Developing pigment dispersion,iris stroma atrophy,transillumination defects and pupil deformation were found in DBA/2J mice.IOP began to rise in 7-month-old DBA/2J mouse,peaked in 9-month-old mouse and returned to normal in 14-month-old DBA/2J mouse.A significant difference was found in IOP among different ages of DBA/2J mice (F=27.600,P＜0.05) but not C57BL/6J mice (F=0.249,P=0.781).RGCs loss was observed in 7-month-old DBA/2J mice and more serious from 9 to 11-month-old mice,showing a significant decline of RGCs among different ages of DBA/2J mice (F=23.594,P=0.000) but not C57BL/6J mice (F=1.816,P=0.211).The abnormality of optic nerve cupping and decrease of retinal nerve fiber layer were found in 9 to 14-month-old months old DBA/2J mice and were normal in age-matched C57BL/6J mice.Conclusion The abnormal alteration of the ocular anterior segment in DBA/2J mouse is gradually aggravated with aging.The findings of eye in DBA/2J mouse is characterized by secondary glaucoma.DBA/2J mouse offer an ideal animal model for the research of glaucoma.

Expression of heat shock protein 27 induced in the experimental rat glaucoma model / 眼科

Objective To evaluate changes in retinal expression of three heat shock proteins(HSPs)HSP27,HSP70 and HSP90 in normal and experimental glaucomatous rats's retina,and to explore the potential relationship between HSPs with glaucoma optic neuropathy.Design Experimental study.Participants 60 Wistar rats.Methods Rats were randomly divided into ocular hypertension group(n=30)and sham control group(n=30).The right eye was designed as the experimental eye,and the left as the control eyes.Intraocular pressure(IOP)was elevated by using underwater bipolar electro coagulation condensate 3 episcleral and limbal veins on the right eye of rats to establish of animal models of glaucoma.IOP were monitored twice weekly with a Tonopen.In treated eyes,IOP ranged from 27 to 35 mmHg throughout the study.Rats were killed with euthanasia at 10,20,or 60 days following the occurrence of ocular hypertension,with retina dissected free and protein isolated.Protein was used for Western blot analysis and probed with specific HSP antibodies,and normalized to ?-actin levels.Main Outcome Measures IOP,Western bolt.Results Western blot analysis demonstrated that HSP27 protein levels in the retinas were elevated as much as 197% at 10,20 and 60 days following the induction of ocular hypertension.No changes in protein levels were observed for HSP70 or HSP90 in retina from ocular hypertensive eyes.Conclusions The stress protein HSP27 is upregulated in retinas from ocular hypertensive rats.No changes in HSP70 or HSP90 were observed.The upregulation in HSP27 appears to be a gene specific event associated with elevated IOP,its expression may be a potential relationship associated with optic neuropathy in glaucoma.