Objective:To detect collagen I synthesis activity in the vitreous of PVR induced by macrophages in rabbits. Methods:PC Ⅲ (Procollagen Ⅲ ) concentrations were measured by radioim- munoassay in the vitreous samples of 14 rabbit eyes with experimental PVR and 14 control eyes. Results:The mean PC Ⅲ concentration on the 7th day after macrophage injection as 257.58mu;g/L(range,236.04~266.88mu;g/L,n= 4)and significantly increased on the 14th day later. On the 28th day the mean concentration of PC Ⅲ as 912.23mu;g/L (range, 881.36~943.10mu;g/L ;n= 2). There was a significant difference between the 7th and the 14th, 21st of 28th day statistically(P＜0.05). PC Ⅲ was not detected in control eyes. Conclusion:The PC Ⅲ level in the vitreous of rabbit eyes with experimental PVR increased significantly from the 7th to the 28th day after macrophages injection and is well consistent with the time course of scarring and the development of traction retinal detachment in the PVR model. （Chin J Ocul Fundus Dis,1996,12: 43-44）
Objective To investigate the effect of hypericin on the activity of protein kinase C (PKC) in cultured human retinal pigment epithelium (RPE) cells in vitro.Methods RPE cells were cultured in standard medium with 10% serum concentrations containing 0.5 to 5.0 μmol/L hypericin with or without preincubation of phorbol 12-myristate 13-acetate (PMA). The activities of cytosolic PKC (c-PKC) and membranous PKC (m-PKC) were assayed by PKC kit. Results The original activities of c-PKC and m-PKC of RPE cells were (35.34±4.10) pmol·min-1·mg-1and (62.52±8.80) pmol·min-1·mg-1.The activity of c-PKC in RPE cells with PMA preincubation decreased rapidly in 5 minutes, with a subsequent slow decrease after 20 minutes and a decrease to 18% of the activity of c-PKC in RPE cells without PMA preinubation after 60 minutes. While the activity of m-PKC in RPE cells with PMA preincubation increased gradually after 5 minutes and reduced after reached the peak at 40 minutes, and then returned to baseline after 60 minutes, eventually decreased below 30% of the control group. When RPE cells were cultured with PMA for 48 hours, the activities of c-PKC and m-PKC were hardly detectable, while RPE cells were cultured with both PMA and hypericin, hypericin could counteract most of down-regulation by PMA. Conclusion Hypericin may inhibit the translocation of PKC in RPE cells,change the activity of PKC, promote the apoptosis of RPE cells likely,and then prevent proliferative vitreoretinopathy. (Chin J Ocul Fundus Dis，2003，19：55-58)
ObjectiveTo observe the longterm effect of suramin on the inhibition of proliferation of human retinal pigment epithelial (RPE) cells in vitro. MethodsRPE cells grown in 9 pieces of 96well plate (12 wells each plate) were divided into experimental and control group, with 6 wells in each group. The concentration of 0.1 ml RPE cells in each well is 5×104 cells/ml. After the change of the medium, RPE cells were treated with suramin (250 μg/ml) in experimental group while treated with nothing in the control group. The medium of the 2 groups were changed to the normal medium after 4 days. At the 1st, 2nd, and 4thday after the addition of suramin and at the 1st, 2nd, 3rd, 5th, 6th, 7th, 9th , 11th and 13th day after removing suramin, 1 plate was randomly selected to stop culturing, and the proliferation of RPE cells were detected by methyl thiazolyl tetrazolium (MTT) assay. ResultsUnder reversed microscope, RPE cells in control group were fused completely at the 7th day after inoculation. The extracellular space of RPE cells in experimental groups was larger than that in the control group, and remained unfused at the 13th day after inoculation. The inhibitory rate of proliferation of RPE cells at the first day after treated with suramin was 14.85% and increased to the highest 25.79% at the 4th day. The first day after the suramincontaining media was removed, the inhibitory rate decreased to 12.35%, and then raised gradually to over 20% at the 3rd to 5th day. Finally, the rate drop to 14.71%. ConclusionSuramin has the long-term effect on the inhibition of RPE cells induced by serum, especially the inhibitive effect after the remove of suramin, which indicates the specific double-peak inhibition during the whole process.(Chin J Ocul Fundus Dis, 2005,21:25-27)
Objective To observe the surgical results of the patients with persistent hyperplastic primary vitreous (PHPV). Methods Clinical data of 16 eyes of 16 patients with PHPV who had undergone retina surgery were retrospectively analyzed. The patient, 7 males and 9 females, aged from 3 months to 25 years with the average of 51.9 months. 14 cases were si n gle eye pathogenesis, 2 cases were binocular pathogenesis. Of the 16 eyes, 3 had anterior PHPV, 13 had components of both anterior and posterior disease. In addition, 10 eyes with cataract, 7 eyes with posterior synechia, 5 eyes with shallo w anterior chamber, 3 eyes with bandshaped degeneration of cornea, 1 eye with corneal opacity, 2 eyes contractive retinal detachment, 1 eye with rhegmatogenous retinal detachment. The visual acuity was light sensation, hand move, counting f ingers/10cm and 002 in 1 eye respectively. 12 eyes can not meet visual inspection, the reflection of b light was not obvious.13 eyes underwent lensectomy and anterior vitrectomy, 1 eye was implanted intraocular lens, 3 eyes with retin al detachment underwent lensectomy, vitrectomy, photocoagulation, gas tamponade and scleral buckling. Follow-up ranged from 6 months to 4 years (mean 15.3 months). Results After surgery, all 16 eyes had normal intraocular pressure and anterior chamber, retinal attachment; two eyes achieved a visual acuity of 10/100 and 20/ 100; 8 patients have the reflection of b light who once didnprime;t meet visual inspection; the visual acuity in 6 patients below counting fingers. Conclusions Surgical treatment of PHPV can prevent future glaucoma and he morrhagic complications. It is useful to win functional visual acuity if it combined with amblyopia training after surgery. （Chin J Ocul Fundus Dis，2008，24：210-212）
ObjectiveTo observe the expression of heat shock protein 47 (HSP47) and transforming growth factor-β2(TGF-β2) in vitreous specimens and epiretinal membranes of patients with proliferative vitreoretinopathy diseases. MethodsVitreous specimens and epiretinal membranes were obtained from 48 patients (48 eyes) with proliferative vitreoretinopathy (PVR) and 50 patients (50 eyes) with proliferative diabetic retinopathy (PDR). Vitreous specimens and internal limiting membranes were collected from 20 patients (20 eyes) with idiopathic macular hole (IMH) as control group. The expression of HSP47 and TGF-β2 in the vitreous specimens was evaluated using enzyme linked immunosorbent assay. The expression of HSP47, TGF-β2, typesⅠandⅢcollagen in epiretinal membrane and internal limiting membrane specimens were observed for immunohistochemical staining method. The correlation between the positive expression of HSP47 and TGF-β2, typesⅠandⅢcollagen in epiretinal membrane specimens of patients with PVR and PDR were analyzed. ResultsThe expression of HSP47 in vitreous specimens of patients with PVR, PDR and IMH were (212.35±23.32), (231.30±26.79), (171.06±28.91) pg/ml, respectively. The expression of TGF-β2 in vitreous specimens of patients with PVR, PDR and IMH were (1919.96±318.55), (1939.39±177.57), (1194.61±234.20) pg/ml, respectively. The expression of HSP47, TGF-β2 in the vitreous specimens of patients with PVR and PDR were significantly increased compared with patients with IMH and the difference was statistically significant (F=12.952, 34.532;P < 0.01). The epiretinal membrane of patients with PVR and PDR showed markedly increased expression of HSP47, TGF-β2, typesⅠandⅢcollagen in the cytoplasm and extracellular matrix. The expression of HSP47 and typeⅢcollagen was negative and the expression of TGF-β2 was weakly positive and the expression of typesⅠcollagen was positive in internal limiting membrane of patients with IMH. The expression of HSP47, TGF-β2, typesⅠandⅢcollagen in the epiretinal membrane of patients with PVR and PDR were significantly increased compared with patients with IMH and the difference was statistically significant (F=13.469, 18.752, 12.875, 20.358; P < 0.01). The expression of HSP47 was positively correlated with the positive expression of TGF-β2, typesⅠandⅢcollagen in epiretinal membrane specimens of patients with PVR (r=0.475, 0.556, 0.468; P < 0.05) and PDR (r=0.484, 0.589, 0.512; P < 0.05). ConclusionsThis study showed increased consistent expression of HSP47 and TGF-β2 in vitreous and epiretinal membrane specimens of patients with PVR and PDR. Both HSP47 and TGF-β2 were expressed in the cytoplasm and extracellular matrix. HSP47 and TGF-β2 may be involved in the pathological process of PDR and PVR by promoting collagen synthesis.
Objective To detect the variation rule of different cellular components, extracellular matrix, matrix-metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases（TIMPs）in proliferative membranes in proliferative vitreoretinopathy (PVR) with different courses of disease, and to investigate the remodeling mechanism of PVR. Methods Sixteen surgically excised specimens of proliferative membranes from patients with rhegmatogenous retinal detachment combined with PVR with the course of disease of 2 months to 8 years were selected. The different cellular component of retinal pigment epithelial (RPE) cells and glial cells, component of extracellular matrix including fibronectin, laminin，and collagen types Ⅰ to Ⅳ, and matrix metalloproteinases (MMP2, MMP9) and TIMP1 in proliferative membrane were labeled by immunohistochemical method. The variati on of those labeled components in proliferative membrane in PVR duration and the correlation between these components and the course of PVR were analyzed. Results As the duration of PVR increased,the expression of RPE cells, fibronectin and MMP2 decreased (Plt;0.05）,while glial cells,collagen type Ⅰ and Ⅲ increased (Plt;0.05）.The positive staining of laminin and collagen type Ⅱ and Ⅳ were found, but the association with PVR duration was not detected. A negative correlation between PVR duration and RPE cells, MMP2, and fibronectin respectively and a positive correlation between PVR duration and glial cells, collagen Ⅰand Ⅲ respectively were detected. MMP2 positively related with variation of fibronect in. Positive staining of MMP9 and TIMP1 was recorded but did not change with the variation of the disease course. Conclusion During the formation and development of proliferative membrane in PVR, RPE cells, glial cells, fibronectin, collagen type Ⅰand Ⅲ and MMP2 take part in the remodeling of proliferative membrane. (Chin J Ocul Fungdus Dis, 2006, 22:308-312)
Objective To observe the effect of medicineinduced posterior vitreous detachment (PVD) on proliferative vitreoretinopathy (PVR). Methods PVR was induced in the left eyes of 24 pigmented rabbits by intravitreal injection with platelet rich plasma. The rabbits were randomly divided into two experimental groups (group A and B) and one control group with 8 eyes in each group. Three hours later, the eyes in group A and B and the control group underwent intravireal injection with 1 U plasmin 0.05 ml+20 U hyaluronidase 0.05 ml, plasmin 0.1 ml, and balance salt solution 0.1 ml, respectively. The grade of PVR was recorded 1, 7, and 28 days after the intravitreal injection, and the eyes were examined by flash electroretinogram (FERG), B-scan, and retinal histopathological examination. Results The PVR models of rabbit eyes were induced successfully. On the 7th day after injection, complete and partial PVD was found in 5 and 3 eyes respectively in group A; partial PVD in 5 eyes and no complete PVD was observed in group B; there was no PVD in the other 3 eyes in group B and also in the eyes in the control group. On the 28th day after intravitreal injection, PVR grade of group A and B were both obviously lower than that of the control group（D=75.6, 98.9；P=0.003，P=0.011）; On the 7th and 28th day after injection, the b-wave amplitude in group A and B was significantly higher than that in the control group; PVR grade of the PVD eyes was lower than that of nonPVD eyes; PVR grade of the complete PVD eyes was only 0～1. Conclusions Three hours after the PVR models of rabbit eyes were induced, complete PVD induced by intravitreal injection of plasmin combined with hyaluronidase could prevent the development of PVR of rabbit eyes in some degree; partial PVD induced by plasmin alone or combined with hyaluronidase could relieve the development of PVR.
PURPOSE:To evaluate the ability Of retinoic acid(RA) in silicon oil(SiO)to inhibit the proliferation of injected intraocular fibroblast cells. METHODS:Thirty New Zealand white rabbits (58 eyes)were divided into three groups. In control group ,only SiO(10 eyes)or BSS(10 eyes)were injected intravitreally and 5mu;g/ml (18 eyes)or 10mu;g/ml (20 eyes)RA in SiO were injected into other lwo groups respectively. Three days after gas-compression vitrectomy, 2 times;105 fibroblasts and Sio(0.5ml)or BSS(0.5ml)were injected in all eyes sequentially.The morbidity of tractional retinal detachment (TRD) were observed by ophthalmoscope until 4 weeks. RESULTS:After 4 weeks,in control ,5mu;g/ml RA in SiO and 10mu;g/ml RA in SiO group,80. 00%,44.44%,and 30.00% eyes developed TRD respectively. Significant statistical differences were found between the control group and the two treated groups (P＜0.05). CONCLUSIONS:5mu;g/ml or 10mu;g/ml RA in SiO can inhibit the occurrence of TRD effectively. （Chin J Ocul Fundus Dis,1997,13:174-176）
ObjectiveTo observe the expression of hot shock protein 47 (HSP47) in pre-retinal membrane of proliferative vitreoretinopathy (PVR) and the influence of transforming growth factor-β2 (TGF-β2) on the expression of HSP47 in retinal pigment epithelial (RPE) cell. MethodsPre-retinal membranes were collected and observed by hematoxylin-eosin, Masson and immunohistochemical staining. Cultured ARPE-19 cells were treated with TGF-β2 at serial concentration (0, 1, 5, 10 ng/ml) and time (0, 12, 24, 48 hours), respectively. And then the mRNA and protein expressions of HSP47 and Col-Ⅰ were measured by fluorescence quantitative reverse transcription polymerase chain reaction and Western blot at the same time. ResultsA lot of epithelial cells with pigmental particles were observed in pre-retinal membranes of PVR, much accumulated collagen protein was observed in the specimens, and HSP47 positive expression was bserved in cytoplasm and stroma of most of the epithelioid cells. Compared with 0 ng/ml group, the expressions of HSP47 mRNA in ARPE-19 were up-regulated by 1.32, 2.35, 1.85 fold, significant differences were observed in all groups (F=27.21, P<0.05); the expressions of protein were up-regulated by 2.33, 2.89, 2.60 fold, significant differences were observed in all groups (F=39.78, P<0.05). The expressions of Col-Ⅰ mRNA were up-regulated by 1.29, 1.52, 2.11 fold, significant differences were observed in all groups (F=23.45, P<0.05); the expressions of protein were up-regulated by 1.18, 1.49, 2.11 fold and significant differences were observed in all groups (F=29.10, P<0.05). Compared with 0 hour group, the expressions of HSP47 mRNA were up-regulated by 1.56, 1.84, 2.86 fold in ARPE-19 cells stimulated by 5 ng/ml TGF-β2 for 12, 24 and 48 hours, and the differences were all significant (F=31.56, P<0.05); the expressions of protein were up-regulated by 2.08, 2.37, 2.80 fold, and the differences were all significant (F=49.18, P<0.05). The expressions of Col-Ⅰ mRNA were up-regulated by 1.57, 1.86, 2.78 fold and the differences were all significant (F=54.43, P<0.05), the expressions of protein were up-regulated by 1.38, 1.59, 2.16 fold and the differences were all significant (F=42.52, P<0.05). ConclusionTGF-β2 may play a role in the pathologic process of PVR by promoting the expression of HSP47 and then increasing the synthesis and accumulation of Col-Ⅰ.