【摘要】 背景:体内实验证实成纤维细胞生长因子能有效保护庆大霉素致肾小管上皮细胞的损伤,但对体外培养细胞的作用如何不多见。目的:在建立庆大霉素肾毒性体外细胞模型的基础上,观察不同浓度碱性成纤维细胞生长因子对庆大霉素肾毒性的保护作用。方法:采用酶加网筛方法分离纯化昆明小鼠肾小管上皮细胞,调整细胞浓度为1×108L-1,将细胞悬液移入96孔细胞培养板,分组培养:空白对照组:正常培养;庆大霉素组:10,30,50μL/孔(即400,1200,2000U/孔)记为G1、G2、G3;碱性成纤维细胞生长因子组:20,50,80μL/孔(即90,225,360ng/孔)记为B1、B2、B3;庆大霉素加碱性成纤维细胞生长因子干预组:先加碱性成纤维细胞生长因子12h后,再加庆大霉素12h培养,分9个剂量组,即G1B1、G1B2、G1B3、G2B1、G2B2、G2B3、G3B1、G3B2、G3B3,每组4复孔。观察细胞形态及数量变化。结果与结论:庆大霉素对肾小管上皮细胞的损伤呈剂量依赖性,中、高浓度组的上皮细胞皱缩,变圆,肿胀,贴壁差,内部胞质破坏严重,结构紊乱,低浓度组细胞数量改变不明显,并且开始有成纤维细胞出现;碱性成纤维细胞生长因子各组细胞饱满、折光性强,数量明显增多,50μL/孔浓度以上效果显著,与80μL/孔差异无显著性意义;庆大霉素加碱性成纤维细胞生长因子干预组中低浓度庆大霉素组细胞未见明显损害,细胞数量反而增多,中浓度庆大霉素组损害的细胞崩解减少、细胞皱缩和贴壁差的程度有所减轻,高浓度碱性成纤维细胞生长因子干预后细胞形态良好,但高浓度庆大霉素所致细胞肿胀、坏死损伤任何浓度的碱性成纤维细胞生长因子干预都无法改善。50μL/孔碱性成纤维细胞生长因子对中、低浓度庆大霉素所致肾毒性有拮抗作用,对高浓度庆大霉素所致肾毒性无保护作用。更多还原

【Abstract】 BACKGROUND:In vivo experiments have confirmed that fibroblast growth factor can effectively protect gentamicin-induced renal tubular epithelial cell injury,but the effect on the in vitro cultured cells is still rare. OBJECTIVE:To explore the mechanisms of basic fibroblast growth factor (bFGF) at different concentrations on preventing nephrotoxity mediated by genamicin on the primarily cultivated renal tubular epithelial cell models. METHODS:By use of enzyme and mesh screening,renal tubular epithelial cells were isolated from Kunming mice and purified,adjusting the cell concentration of 1×108/L,then cell suspension was moved to a 96-well culture plate and divided into different groups for culture:blank control group:normal culture; gentamicin group:10,30,50 μL/hole (ie,400,1 200,2 000 U/holes) are recorded as G1,G2,G3; bFGF group:20,50,80 μL/hole (ie,90,225,360 ng/hole) are recorded as B1,B2,B3; gentamicin plus bFGF intervention group:after adding bFGF 12 hours,then added gentamicin 12 hours,assigned into 9 dose subgroups,namely,G1B1,G1B2,G1B3,G2B1,G2B2,G2B3,G3B1,G3B2,G3B3,each subgroup contained four-hole complex. Cell morphology and quantity was observed. RESULTS AND CONCLUSION:Gentamicin showed a dose-dependent effect on the renal tubular epithelial cell injury,epithelial cells in the medium and high concentration groups exhibited shrinkage,rounded,swelling,poor adhesion,severely damaged cytoplasm and structural disorder. In the low concentration group,the number change of cells was not obvious,and fibroblasts began to appear; In the bFGF groups,cells were full,exhibited strong refraction,the cell number increased significantly,these manifestations were significant in 50 μL/hole concentration,and there was no significant difference compared with 80 μL/hore concentration; in case of gentamicin plus bFGF intervention,cells with low concentrations of gentamicin had no obvious damage to cells,which increased,the damaged cells collapse was reduced in the group of low concentration of gentamicin,cell shrinkage and poor adhesion were slightly relieved,high concentrations of bFGF intervention could yield to good cell morphology,but high concentrations of gentamicin caused cell swelling and necrosis of injury,which could not be improved by any concentrations of bFGF intervention. 50 μL/hole bFGF has antagonistic effect on the nephrotoxicity mediated by medium and low concentrations of gentamicin,but has no protection on high concentration of gentamicin-induced nephrotoxicity.更多还原