Objective:To observe the changes of the thickness of reti nal nerve fiber layer (RNFL) of optic disc in rats with chronic glaucoma continuously dete cted by optic coherence tomography (OCT). Methods:A total of 48 Wist ar rats (24 males and 24 females) were randomly divided into 3 groups with 16 ra ts (32 eyes) in each group. The right eyes were the photocoagulation eyes and the left ones were as the control. Laser photocoagulation with the wavelength of 532 nm was perfo rmed on the trabecular network of the right eyes to induce the chronic middlelevel oc u lar hypertension. The changes of the intraocular pressure (IOP) were observed. O pticdisc linear scanning of OCT was performed 3, 6, and 9 weeks after IOP incr e ased, and the thickness of RNFL of optic disc was detected by the computer. Eight rats in each group were killed and retinal histology slic es were used to detect the thickness of RNFL. The flatmount s of retina from the right eyes of the other 8 rats in each group were stai ned by 1% toluidine blue. The density of retinal ganglion cells (RGC) was calcul ated and the results were compared and analyzed. Results:IOP o f the rats increas ed chronically and moderately after photocoagulation. IOP of the experimental ey e 3,6, and 9 weeks after photocoagulation was obviously higher than which of the control eyes, respectively （P＜0.001）. The results of OCT showed that the thickness of the RNFL of the experimental eyes was (67.39plusmn;5.91) mu;m, (53.4 2plusmn;5.64) mu;m，and (44.35plusmn;5.76) mu;m 3, 6, and 9 weeks after photocoagulation, and the corresponding thickness in the control eyes was（80.32plusmn;5.87）, （79.69plusmn;5.69）, and （80.78plusmn;5.84）mu;m, respectively. The thickness of the retinal fiber layer detecte d by histological method was （64.38plusmn;6.54）, （51.47plusmn;6.4）, and （42.10 plusmn;6.10）mu;m in the experimental eyes 3, 6, and 9 weeks after photocoagulation, and (76.23plusmn;6.78), (78.64plusmn;6.15), and (77.64plusmn;6.63) mu;m in the control eyes. Regression analysis of the thickness detected by the two methods was made, and the regression coefficients was 0.932（P＜0.001）.The differ ence of the ave rage density of RGC between the two groups was significant (P＜0.05）. Conclusi on:Glaucoma model in Wistar rats may successfully set up b y photocoagulating the trabecular meshwork. The thickness of retinal nerve fiber layer of the optic disc in rats with chronic glaucoma detected by OCT and obser ved by the light m icroscope is accordant. The changes of the thickness of RNFL in rats with chroni c glaucoma could be continuously detected by OCT to investigate the progress of the glaucomatic retinopathy in rat model.
The occurrence of high intraocular pressure (IOP) after vitrectomy for diabetic retinopathy (DR) is related to many factors, including the type and stage of DR, macular detachment, surgical methods, and the type of ocular tamponade. Early high IOP occurred mainly due to laser photocoagulation, inflammatory response, improper ocular tamponade, residual viscoelastic agents and ciliary body dysfunction. In addition to the above reasons, early-middle stage high IOP is also related to tamponade gas expansion peak, encircling scleral buckle and hyphema. The major reason for middle-stage high IOP is hyphema and silicon oil in anterior chamber. The reasons for late-stage high IOP are glaucoma, silicone oil emulsification, long-term use of glucocorticoid, and iris incision closure. Most high IOP can be controlled by proper treatment such as stopping use of glucocorticoid, anti-glaucoma eye drops and surgeries. But there are still a small number of patients with unexplained refractory high IOP, the mechanism need to be further explored.
Objective To investigate the effect of proanthocyanidins (PC) on retina of rats with acute ocular hypertension. Methods The SpraqueDawley (SD) rats were randomly divided into the normal group, the model group and PC high or lowdosage group. The PC high or low dose group received 300mg/kg.d or 100mg/kg.d of PC in suspension solution for 5 days respectively. The normal group and the model group fed with distilled water for 5 days. Then acute ocular hypertension was induced in the model group and all PC groups, and after 48h of ocular hypertension the eyeballs were analyzed by electron microscope and UV spectrophotometer to measure the levels of superoxide dismutase(SOD), malonaldehyde (MDA), nitrogen monoxidum (NO), glutamic acid (Glu) and calcium ion(Ca2+). Results PC could raise the activity of SOD and reduced the levels of MDA,NO,Glu and Ca2+ in retina tissue. Electron microscope examination revealed that PC reduced retinal edema and ganglion cell apoptosis. PC also enhanced the SOD activity and suppressed the levels of MDA, NO, Glu and Ca2+. Conclusions PC can protect retina from acute ocular hypertension. The main mechanism might relate to anti-free radical oxidation, antagonizing calcium overloading, reducing toxicity of NO and Glu on the retina.
ObjectiveTo investigate the effect of erigeron breviscapus (EBHM) on ocular hypertension and the protective effect of retinal ganglion cells (RGCs) in rats by regulating mitogen activated protein kinase (MAPK) signaling pathway.MethodsSixty male Sprague-Dawley rats were divided into control group, model group, low-dose EBHM group (group A), medium-dose EBHM group (group B), and high-dose EBHM group (group C) by random number table method. There were 12 rats in the group, the left eye was used as the experimental eye. The rats of model group, group A, group B, and group C were infused with normal saline through the anterior chamber to construct an acute ocular hypertension model; the control group was given general anesthesia only. Then, 2-30 days after modeling, rats in the control group and model group were given 3 ml of normal saline once a day; rats in group A, group B, and group C were given 0.30, 0.45, and 0.60 g/100 g EBHM by intragastric administration, respectively, 1 time/d. The rat intraocular pressure was measured before modeling and 1, 14, and 30 days after modeling, and the proportion of high intraocular pressure model was measured. Thirty days after modeling, hematoxylin-eosin (HE) staining was used to observe the pathological changes of retinal tissue; immunofluorescence staining was used to detect the changes in the number of RGCs; real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to detect p38 in the retinas of rats in each group. The relative expression of MAPK and Caspase-3 mRNA; western blot was used to detect p38MAPK and phosphorylation in the retina of rats in each group relative expression of phosphorylate-p38MAPK (p-p38MAPK) and Caspase-3 protein. One-way analysis of variance was used for multi-sample comparison, and SNK-q test was used for comparison between two samples.ResultsOne day after modeling, none of the rats in the control group developed acute ocular hypertension, and the other groups were successfully modeled. Compared with the model group, the rates of acute ocular hypertension at 14 days after modeling in groups B and C were lower (χ2=98.701, P＜0.05), and the rates of acute ocular hypertension at 30 days after modeling in groups A, B, and C were 0. There was no statistically significant difference in the rates of acute ocular hypertension between 14 and 30 days after modeling in the A, B, and C groups (P＞0.05). The results of HE staining showed that the structure of the retina in the control group was complete, and the layers were clearly visible; the RGCs count was not abnormal, and the morphology was plump and round. The retina of rats in the model group became thinner; the number of RGCs was greatly reduced, the morphology was vacuolated, and the arrangement was sparse. The retina of rats in groups A, B, and C became thicker, and the number of RGCs increased, and the retina structure in group C was better restored. The results of immunofluorescence staining showed that the RGCs counts of rats in groups A, B, and C were higher than those in the model group, and the difference was statistically significant (F=297.514, P＜0.05); pairwise comparison between groups, group A was lower than that of group B and C Group (q=2.842, 5.263), group B was lower than group C (q=2.457), the difference was statistically significant (P＜0.05). The results of RT-qPCR and Western blot showed that compared with the model group, the relative expression of Caspase-3 mRNA (F=267.912) and protein (F=692.279) and the relative expression of p-p38MAPK protein in the retina of rats in groups A, B and C. The expression level (F=150.061) all decreased, and the difference was statistically significant (P＜0.05); pairwise comparisons between groups showed that Caspase-3 mRNA (q=6.977, 15.642) and protein (q=6.997, 15.642) relative expression levels and p-p38MAPK protein (q=12.443, 24.358) relative expression levels are lower than groups A and B, group B was lower than group A (q=11.678, 12.471, 10.204), the difference was statistical academic significance (P＜0.05).ConclusionsEBHM can significantly reduce intraocular pressure in rats with acute ocular hypertension, increase RGCs counts, and reduce retinal damage. Its regulatory mechanism may be related to the MAPK pathway.
Objective To observe the clinical characteristics of steroid-induced ocular hypertension (SIOH) in patients with uveitis, and explore the relationship between its clinical phenotype and gene polymorphism. Methods A retrospective case-control study. From July 2019 to December 2020, 576 patients with uveitis who were treated with glucocorticoid eye drops in Tianjin Medical University Eye Hospital were included in the study. Among them, there were 175 confirmed glucocorticoid responders (SRs) and 401 glucocorticoid non-responders (NRs). Seventy cases of SRs (age ≥18 years) using 1% prednisone acetate eye drops were selected as the experiment group and 64 cases of NRs were selected as the control group. The polymorphism of rs2523864 and rs3873352 of human leukocyte antigen complex group (HCG) 22 gene were detected by Sanger sequencing. To observe the clinical characteristics of SIOH after the use of glucocorticoid eye drops, and the correlation between rs2523864 and rs3873352 and the occurrence of SIOH. Differences among groups were compared with the Chi-square test or Fisher's exact test. The correlation between the occurrence of SIOH and the range of intraocular pressure increases after glucocorticoid use and the rs2523864 and rs3873352 loci were compared using the odds ratio (OR) and its 95% confidence interval (CI). Results SIOH occurred in 175 (30.4%, 175/576) of 576 patients. Among them, there were 96 males (54.9%, 96/175) and 79 females (45.1%, 79/175); the average age was 33.64±17.40 years. Steroid high responders (HRs) and steroid moderate responders (MRs) were 58 (33.1%, 58/175) and 117 (66.9%, 117/175) cases. The medication time for the increase in intraocular pressure in MRs that was 33 (19, 56) days, and in HRs that was 28 (14, 36) days, the difference of which was significant (Z=－1.999, P=0.046). No differences were found in daily doses of ocular hypertension induced by 1% prednisone acetate eye drops between MRs which was 4.24 (3.46, 4.66) drops/day and HRs that was 4.32 (3.84, 5.36) drops/day (Z=－1.676, P=0.094). The genotype and allele frequency distribution of the rs3873352 locus in the case group and HRs group were significantly different from those in the control group (P＜0.05). The intraocular pressure with rs3873352 GG genotype after the medication was higher than that with GC and CC genotype (Z=2.855, 2.628; P=0.013, 0.026), whereas there was no significant difference between different genotypes of rs2523864 (Z=3.580, P＞0.05). Genetic model analysis revealed the risk of SIOH in rs3873352 G allele carriers (GG+GC) was 2.048 times that of non-G allele carriers (OR=2.048, 95%CI: 1.027-4.081, P=0.041). The genotype and allele frequency of rs2523864 locus showed no significant difference between different group (P＞0.05). Conclusions After the use of glucocorticoid eye drops, HRs have an earlier increase in intraocular pressure than MRs. HCG22-rs3873352 gene polymorphism is related to the occurrence of SIOH, GG genotype increases the risk of SIOH, and G allele is a risk gene for SIOH.
Objective To analyze the protective effects of heat-shock response on the retinae of the rats after retinal ischemic reperfusion injury.Method Twenty Wistar rats (20 eyes) were divided into 4 groups: intracameral perfusion group (group P), intracameral perfusion after quercetin injection group (group P+Q), intracameral perfusion after heat shock group (group P+H), and in tracameral perfusion after quercetin injection and heat shock group (group P+Q+H ). According to the standard program established by International Society for Clinical Visual Electrophysiology, we recorded the results of the dark-adapted electroretinogram (D-ERG ),oscillatory potentials (OPs),and light-adapted ERG (L-ERG) of the rats with intraocular hypertension after induced by heat shock response. The expressions of HSP 70 of the rats in all groups were observed by Western blotting.Results The expression of HSP 70 of the rats in group P+H was the highest in all groups, but the expressions of HSP70 in group P+Q and P+Q+H were inhibited significantly. The amplitudes of a and b wave of ERG and O2 wave of OPs decreased, and the delitescence of them were delayed significantly in rats after intracameral perfusion. The amplitude of b wave of D-ERG and O2 wave of OPs in group P+H were higher than which in group P. Zero hour after perfusion, the amplitudes of all waves in group P+H increased significantly (Plt;0.05). Twenty-four hours after perfusion, the retinal functional resumption of the rats in group P+H was better than which in group P. In group P+Q and P+Q+H, the delitescences of all waves of ERG and O2 wave of OPs were the longest and the amplitudes were the lowest, and some waves even disappeared.Conclusions The heat-shock response may improve the recovery ability of the retinal cells after injury of ischemic reperfusion.(Chin J Ocul Fundus Dis，2003，19：117-120)
At present, there are few in vivo experimental studies on anterior chamber flow field, and the relevant technologies are not mature. This study explores the experimental method and key techniques of particle image velocimetry (PIV) for the in vivo measurement of anterior chamber flow field with slow flow velocity in the rabbit with acute intraocular hypertension. The experimental process can be divided into three parts: model construction of rabbit eye with acute intraocular hypertension, in vivo eyeball preparation, and PIV setup. The following key techniques were mainly investigated: the optimal injection strategy of fluorescent particles and the correction strategy for image acquisition errors caused by the effects of image refraction and respiration. The results showed that the best injection method was that 15 μL of fluorescent particles solution was slowly injected into the anterior chamber through the lower part of iris and then the rabbit was released and waited for 13 h. In this way particles were completely distributed in the anterior chamber with the help of the aqueous humor circulation, and then in vivo PIV experiment could be performed. The eyeball should be covered with a square flume filled with ultrasonic coupling gel for the sake of imaging during the experiment. The Maximal Information Coefficient algorithm could be applied to correct the measured results before post-processing calculation. The results indicated that feasible injection strategy of fluorescent particles and the correction strategy for image acquisition are critical to obtain nice experiment effects for the in vivo PIV measurement of anterior chamber flow field in the rabbit with acute intraocular hypertension.