Objective To investigate the effects of glycation pro ducts on growth and cytosoic free calcium ([Ca2+]i) of bovine retinal capillary pericytes. Methods The changes of growth and [Ca2+]i of bovine retinal pericytes,which were cultured in early glycation products of bovine serum albumin (EG-BSA) and advanced glycation end products of bovine serum albumin (AGE-BSA),were studied by counting cell numbers,MTT colorimetric assay,[3H]thymidine incorporating,and fluorescent indicator fura-2 acetoxymeth1 ester (Fura-2AM). Results The number of alive pericytes in groups of EG-BSA and AGE-BSA were 17.87plusmn;2.36 and 14.77plusmn;3.72 which comparing with their control groups (20.54plusmn;0.82 and 20.31plusmn;0.93)were de creased 13.00% and 27.00% (Plt;0.01) by counting cell numbers on a counting plate after four days.The results were 0.4619plusmn;0.0946 and 0.3884plusmn;0.1031 which comparing with their control groups (0.5236plusmn;0.0539 and 0.5227plusmn;0.0519)were decreased 12.00% and 25.70% (Plt;0.01) by MTT colorimetric assay.Amount of [3H]thymidine incorporating in groups of EG-BSA and AGE-BSA were 39450.16plusmn;887 0.68 and 33667.85plusmn;10581.70 which comparing with their control groups (56373.63plusmn;2317.97 and 56542.04plusmn;1961.23)were decreased 30.00% and 40.40% (Plt;0.01).The [Ca2+]i concentration of pericytes in groups of EG-BSA and AGE-BSA were (129.55plusmn;30.41) nmol/L and (179.71plusmn;56.69) nmo l/L which comparing with their control groups [(79.70plusmn;6.94) nmol/L and (83.plusmn;6.39) nmo l/L] were increased to 163.00% and 214.00%. Conclusion Both EG-BSA and AGE-BSA can inhibit the proliferation and DNA syntheses of retinal capillary pericytes,and increased [Ca2+]i concentration in pericytes,especially the AGE-BSA. (Chin J Ocul Fundus Dis,2000,16:139-212)
Objective To investigate the effects of QUE on proliferation and DNA synthesis of cultured retinal pigment epithelium(RPE) cells with or without EGF. Methods With or without EGF, cultured RPE cells were treated with QUE by various concentrations(200,100,50,1mu;mol/L) and with QUE 200mu;mol/L at different times(24-168 hr), cells proliferation and DNA synthesis were evaluated by cell count method and the uptake of thymidine. The viability of cells was determined by trypanblue exclusion. Results The best concentration of QUE which inhibits proliferation and DNA synthesis of PRE cells was 200mu;mol/L. The significant inhibition effect of QUE occurred at 48hr, and the best inhibition of QUE occurred at 96hr. QUE had more powerful effect of antiproliferation on RPE cells, and the viability of RPE cells was over85%. Conclusion The results suggested that QUE could inhibit the proliferation of RPE cells in a dose-dependent and time-dependent manner, especially inhibit the proliferation induced by EGF stimulating. QUE had no cyto-toxic effect on RPE cells cultured in vitro. (Chin J Ocul Fundus Dis,1999,15:27-29)
Porpose To investigate the optimal concentration of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) on DNA synthesis and their synergism indensity arrested human retinal pigment epithelial (RPE) cells. Methods Growth factor effects in cultured human RPE of the 6th generation were assessed by [3 H]-thymidine incorporation and radioautography. Results EGF and bFGF were potent stimulators when used alone,and their optimal concentrations were 10ng/ml in DMEM and 1ng/ml in 2% serum DMEM.When used in combination (10ng/ml EGF and 10ng/ml bFGF),they caused a significant enhancement of [3 H]-thymidine incorporation about 2.96 times. Conclusion EGF and bFGF were potent stimulators in RPE cells,and demonstrated synergism in their action. (Chin J Ocul Fundus Dis,1998,14:98-100)