糖基化产物对牛视网膜微血管周细胞增生和胞浆[Ca^(2+)]i水平的影响

Effects of glycation products on growth and cytosolic free calcium in bovine retinal capillary pericytes

目的 探讨糖基化产物对牛视网膜周细胞 (pericyte ,PC)增生和胞浆 [Ca2 +]i水平的影响。 方法 采用细胞计数结合噻唑蓝比色测定 ,氚标胸腺嘧啶核苷 (3H thymidine ,3H TdR)掺入和钙荧光探剂 (Fu ra 2Acetoxymethylester,Fura 2AM) ,研究PC在牛血清白蛋白早期糖基化产物 (earlyglycationpro ductsofbovineserumalbumin ,EG BSA)和牛血清白蛋白糖基化终产物 (advancedglycationendproductsofbovineserumalbumin ,AGE BSA)培养下 ,PC增生数、DNA合成量及胞浆 [Ca2 +]i水平的改变。 结果 培养 4d后 ,细胞计数法 :EG BSA和AGE BSA组PC数分别为 17.87± 2 .36 ,14 .77± 3 .72 ,较其对照组 (2 0 .5 4± 0 .82 ,2 0 .31± 0 .93)减少13 .0 0 %和 2 7.0 0 % (P <0 .0 1) ;MTT法 :EG BSA和AGE BSA组分别为 0 .46 19± 0 .0 946 ,0 .3884± 0 .10 13 ,较其对照组 (0 .5 2 36± 0 .0 5 39,0 .5 2 2 7± 0 .0 5 19)减少 12 .0 0 %和 2 5 .70 % (P <0 .0 1) ;3H TdR掺入量 :EG BSA和AGE BSA组分别为 3945 0 .16± 8870 .6 8,336 6 7.85± 10 5 81.70 ,较其对照组 (5 6 373 .6 3± 2 317.97,5 6 5 42 .0 4±196 1 2 3)减少 30 .0 0 %和 40 46 % (P <0 0 1) ;胞浆 [Ca2 +]i水平 :EG BSA和AGE BSA组分别为 (12 9.

Objective To investigate the effects of glycation products on growth and cytosoic free calcium ([Ca 2+ ]i) of bovine retinal capillary pericytes. Methods The changes of growth and [Ca 2+ ]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 17.87±2.36 and 14.77±3.72 which comparing with their control groups (20.54±0.82 and 20.31 ±0.93) were decreased 13.00% and 27.00% (P<0.01) by counting cell numbers on a counting plate after four days.The results were 0.4619±0.0946 and 0.3884±0.1031 which comparing with their control groups (0.5236±0.0539 and 0.5227±0.0519) were decreased 12.00% and 25.70% (P<0.01) by MTT colorimetric assay.Amount of [ 3H]thymidine incorporating in groups of EG-BSA and AGE-BSA were 39450.16±8870.68 and 33667.85±10581.70 which comparing with their control groups (56373.63±2317.97 and 56542.04±1961.23) were decreased 30.00% and 40.40% (P<0.01).The [Ca 2+ ]i concentration of pericytes in groups of EG-BSA and AGE-BSA were (129.55±30.41) nmol/L and (179.71± 56.69) nmol/L which comparing with their control groups [(79.70±6.94) nmol/L and (83.96±6.39) nmol/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 [Ca 2+ ]i concentration in pericytes,especially the AGE-BSA.