Purpose To determine the effect of exogenous interleukin-1alpha; (IL-1alpha;) on the retina and its vasculature and VEGF expression in SD rats． Methods IL-1alpha;2．0 ng (20 mu;l) were injected into the vitreous of 8 left eyes of 8 SD rats while steriled PBS were injected into 8 right contralateral eyes of the same rats as control． All eyes were assessed by direct ophthalmoscopy every day and enucleated on the 7 thpostoperative day． Histological examination (hemato xylineosin staining) and immunohistochemical staining with antibody against VEGF antigen were performed， and sections were observed and photographed under light microscopy． Results ①All 8 IL-1alpha; inject ed eyes developed epiretinal membranes and extraretinal neovascularization on the 3 rd postoperative days while none of the 8 control eyes exhibited any a bnormal retinal vascular changes and they were confirmed by HE staining;②Immuno staining identified VEGF express mainly in the inner layer of vessel walls， the epiretinal membranes， the neuroganglional layer and the photoreceptor layer of retina， while the control eyes showed only weak positive staining in the photo receptor layer． Conclusions IL-1alpha; is capable of inducing vitreo retinal neovascularization，and increasing the expression of VEGF in the retina and epiretinal membranes． (Chin J Ocul Fundus Dis, 2001,17:135-137)
Objective To verify the effect of Evans blue dye on determining the retina blood vessel leakage． Methods Male Sprague-Dawley rats were used in this study． The VEGF induced retinal blood vessel leakage was checked with Evans blue dye． Then the bloodretina barrier breakdown of 1 week diabetic animals was quantified with Evans blue．The dye was extracted from retina by formamide and the extraction was checked with spect rophotometer． Evans blue leakage was normalized against wet or dry retina weight． Results The retinal Evans blue content of eyes treated with VEGF was remarkably higher than that of the controls (n=17 ,Plt;0．0001)． And the eyes of 1 week diabetic duration animals had more Evans blue dye than that of the normal controls (Plt;0．05)． Conclusion Evans blue dye is a sensitive tracer in quantitatively diagnosing the blood retina barrier breakdown． (Chin J Ocul Fundus Dis, 2001,17:221-223)
Objective To determine the effect of thiazolidinediones (TZDs) on early retinopathy in rats with experimental diabetes. Methods In 40 rats, diabetic models were set up in 36 by one-off intraperitoneal injection with streptozotocin (STZ), and other 4 were in the normal control group. Twenty-four diabetic rats with the disease-duration of more than 6 months underwent intravitreous injection (with rosiglitazone or pioglitazone in 10 rats, respectively), and the rest 4 rats werenprime;t injected with drugs as the diabetic positive control group. Immunohistochemical treptomycin-avidin-biotin-complex (SABC) method, in situ hybridization of retinal vascular endothelial growth factor (VEGF) mRNA, and TdT-dUTP terminal nick-end labelling (TUNEL) were performed on the ocular paraffin section to detect the cellular apoptosis. The difference of VEGF expression and cellular apoptosis between TZDs and control group was observed and analyzed. Results The results of immunohistochemical staining and hybridization in situ were negative in the normal control group. The positive expression rate of VEGF was lower in rosiglitazone and pioglitazone group than which in the diabetic positive control group, and there was no obvious differences of positive expression of VEGF mRNA and cellular apoptosis between the 2 groups. Conclusion TZDs (rosiglitazone and pioglitazone) may inhibit the positive expression of VEGF protein in retina of STZ-induced diabetic rats to some extent, but not affect the growth of VEGF in retina. (Chin J Ocul Fundus Dis, 2006, 22: 7-10)
Objective To observe the expression of p53, bcl-2 genes, vascular endothelial cell growth factor（VEGF）, basic fibroblast growth factor（bFGF）, insulin-like growth factor-I （IGF-I）, and the receptors of these factors of retinal vascular endothelial cells (VECs) of 1- to 20-week diabetic rats, and the relationship between the expressions and cell cycle arrest.Methods Retinal sections of diabetic rats induced by alloxan were immunohistochemically stained and observed by light microscopy (LM) and electron microscopy (EM). Dot blotting and Western blotting were used to determine the expression of mRNA, proteins of p53 and bcl-2. Results Under LM, immunohistochemical positive expression of p53 and bcl-2 were found on the vessels of ganglion cell layer and inner nuclear layer of retinae of 8- to 20-week diabetic rats; under EM, these substances were observed depositing in VECs. The retinal VECs also expressed VEGF, bFGF, IGF-I and their receptors. There was no positive expression of other cell types in these retinae, all cell types of retinae in control group, or all cells of retinae of diabetic rats with the course of disease of 1 to 6 weeks. The result of dot blotting revealed that retinal tissue of 20-week diabetic rat expressed p53 and bcl-2 mRNA, and the result of Western blotting revealed that they also expressed p53 and bcl-2 proteins. But retinal tissues of control group did not. Positive expression of bax was not found in the retinae in control group or 1- to 20-week diabetic rats. Conclusion p53, bcl-2 may introduce cell cycle arrest of VECs of retinae in 8- to 20-week diabetic rats. High glucose might stimulate the expression of VEGF, bFGF, IGF-I and their receptors, and the growth factors may keep VECs surviving by self-secretion. (Chin J Ocul Fundus Dis，2003，19：29-33)
ObjectiveTo observe the expression of connective tissue growth factor (CTGF) in injured model of retinal pigment epithelial (RPE) cells and the promoting effect of CTGF on migration of RPE cells.MethodsCultured monolayer-confluent human RPE cells were scraped with a trephine and a cotton stick, and set up the injured model of RPE cells with round scraped area. Immunohistochemistry and in situ hybridization(ISH) were used to detect the expression of CTGF protein and mRNA in injured RPE cells at distinct time points after injury. The number of RPE cells migrated to injured area was measured and the effect of CTGF on migration of RPE cells and the effect of dexamethasone (DEX) on the promoting process of CTGF were observed.ResultsThe results of immunohstochemistry and ISH indicated the weak positive expression of CTGF in RPE cells at the edge of scrape 6 hours after injury, and the positive expression increased gradually as time goes by after the injury. Strong positive expression of CTGF in RPE cells at the edge of scrape was found 24 and 48 hours after injury. Rebuilt human CTGF stimulated migration of RPE cells in a dose-depended manner, and DEX significantly inhabited the migration.ConclusionCTGF involves in the procedure of repair of injury of RPE cells, which may play an important role in the pathogenesis of intraocular proliferative diseases such as proliferative vitreoretinaopathy.(Chin J Ocul Fundus Dis, 2005,21:306-309)
Objective To observe the effects of high concentr at ion glucose on the calcium-activated potassium channel of rabbits′ retinal Müller cells. Methods The rabbits′retinal Müller cells were cultured in vitro under the condition of high concentration glucose, and identified by immunohistochemical staining and transmission electron microscopy. Patch-clamp technique was used to observe the changes of the calcium-activated potassium channel of retinal Müller cells caused by high concentration glucose at different time.Results High concentration glucose could inhibit the calcium-activated potassium channel of cultured retinal Müller cells in a time-dependent manner. Conclusion High concentration glucose may reduce the biological functions of Müller cells by inhibiting calcium-activated potassium channel. (Chin J Ocul Fundus Dis，2003，19：164-167)
Objective To investigate the relationship between the expression of vascular endothelial growth factor(VEGF)and the retinoblastome(RB)differentiation degree and the infitration capability. Method The VEGF expression in RB tissues of 40 cases was analysed by using LSAB immunohistochemical method. Results The VEGF expression in differentiated RB tissues of 13 cases was markedly lower than that in non-differentiaed RB tissues of 27 cases(P＜0.05);The VEGF expression in RB tissues of the optic nerve infiltrated group(14 cases) was significantly higher than of the optic nerve noninfiltrated group(26 cases)(P＜0.05)． Conclusion The results indicate that the VEGF expression is signficantly related with the differentiation degree and infiltration capability of RB. (Chin J Ocul Fundus Dis, 1999, 15: 238-240)
Objective To investigate the effects of Hep-A and Hep-B on vascular endothelial growth factor (VEGF)-induced breakdown of blood-retinal barrier. Methods The mice were subcutaneously injected vehicle, Hep-A or Hep-B 10 mg/kg twice a day for 5 days. Then, 1 μl of 10-6mol/L VEGF were intravitreous injected. After 6 hours, 13.7×104Bq/g3H-mannital were injected intraperitoneally. The mice were sacrificed and the retinas, lungs, kidneys were removed and examined for radioactivity. The result were analyzed using SPSS software to calculate and compare retina/lung and etina/kidney leakage ratio among groups of different treatment. Result The retina/lung and retina/kidney leakage ratio were 0.38±0.04 and 0.21±0.03 respectively in normal mice; increased significantly to 1.05±0.11 and 0.46±0.04 respectively in model mice, both Plt;0.01 compared to those in normal mice; decreased to 0.59±0.06 and 0.32±0.03 respectively in mice treated with Hep-A, both Plt;0.01 compared to those in model mice; decreased 0.54±0.04 and 0.35±0.03 in mice treated with Hep-B,both Plt;0.01 compared to those in model mice. Conclusion Hep-A and Hep-B can significantly reduce VEGF-induced breakdown of blood-retinal barrier in mice. Chin J Ocul Fundus Dis，2004，20：352-354)
Objective To investigate the spatial and temporal regulation effect of VEGF on human fetal retinal vascularization and angiogenesis. Methods The posterior segmental retinas from 54 human fetuses of the 9th week to the 40th week were studied by immunohistodhemistry standing for the expressions of VEGF and PCNA. Results 1. The distribution of VEGF espression was spiking and the peaks were during the 9th-13th and around the 26th week. 2. PCNA immunoreactivity was localized in spindle cells and vascular endothelial cells. The expression level was fluctuated during the developmental process. The peaks were during the 9th-13th and around the 21st week. In these periods, the spindle cells kept proliferating and differentiating, and remodelled subsequently to form the inner side retinal vessels. From the 26th or 34th week, the PCNA immununoreactivity is fully expressed in the vascular endothelial cells of the inner and outer margin of inner nuclear layer(INL) and kept to full terms. 3. Significant positive correlation were shown between the content of VEGF in the retina and that of PCNA in spindle cells and vascular endothelial cells(r=0.736,p＜0.01). Conclusion VEGF was positively involved in modulating human fetal retinal vascularization and angiogenesis. (Chin J Ocul Fundus Dis,1999,15:12-15)
Objective To investigate the effect of batroxobin on the blood-retinal barrier (BRB) and vascular endothelial growth factors (VEGF) in diabetic rats. Methods Sixty Sprague-Dawley rats were used to establish diabetic models by intraperitoneal injecting with streptozotocin (60 mg/kg), and were divided into 3 groups: diabetic group (n=20), batroxobin (40 mg/kg) group (n=20) and batroxobin (20 mg/kg) group (n=20). Twenty-five else rats were in control group. All of the rats were executed 7 days later. The function of BRB was observed by Evans blue method. Results concentration of VEGF protein was detected by enzyme-linked immunoabsorbert assay (ELISA). The results of each group were compared. Results The content of BRB leaked into retina was obvious lower in the control group than which in the other 3 diabetic groups（Plt;0.01）. There was no significant difference of the content of Evans blue between the two groups with different dosage of batroxobin (P＞0.05). The content of Evans blue was lower in the 2 diabetic groups with different dosage of batroxobin than which in the control group (Plt;0.05). The content of VEGF in retina was obviously lower in control group and 2 diabetic groups with different dosage of batroxobin than which in the diabetic group (Plt;0.01), and obviously lower in batroxobin (40 mg/kg) diabetic group than which in the control group (P＝0.01). The content of VEGF in control group and batroxobin (20 mg/kg) diabetic group (P＝0.06) didnprime;t differ much, which occurred similarly in batroxobin diabetic groups with different dosage (P＝0.78). Conclusions Batroxobin may alleviate the damage of function of BRB in diabetic rats and reduce the expression of VEGF, which suggests that batroxobin can protect the function of BRB to a certain extent. (Chin J Ocul Fundus Dis, 2006, 22: 16-19)