摘要
目的 :探讨L ( ) 表没食子基儿茶素没食子酸酯 (L epigallocatechinsgallicacidester,L EGCG)对大鼠肾小球系膜细胞 (glomerularmesangialcells ,GMCs)增生影响及可能的机制。方法 :首先应用不同剂量 (1 0 0 ,2 0 0 ,5 0 0mg/L)EGCG观察其在不同时间 (2 4 ,4 8,72h)对GMCs增生的影响 ,筛选出EGCG对GMCs增生影响的最佳剂量与最佳时间 ;在此基础上 ,将GMCs分为LPS组、对照组、EGCG组、激素组与激素 +EGCG组分瓶培养 ,实验末收集培养细胞 ,涂片 ,应用免疫组化法检测GMCs的增生细胞核抗原 (proliferatingcellnuclearantigen ,PCNA)阳性表达率 ,TUNEL(terminaldeoxynucleotidyltransferasemediateddUTPnickendlabeling,TUNEL)法检测GMCs有效凋亡率 ;应用生化法对培养上清进行 OH ,MDA(malonydialdehyde ,MDA)及SOD (superoxideDismutase ,SOD)活性测定。结果 :2 0 0mg/LEGCG对GMCs增生的抑制效果在 4 8h最佳 ;EGCG组细胞上清中 OH ,MDA较LPS组降低 (P <0 .0 1 ) ,SOD活性较LPS组增高 (均P <0 .0 1 ) ;EGCG组培养细胞PCNA阳性率低于脂多糖组 ,有效凋亡率高于脂多糖组 (P <0 .0 1 )。PCNA阳性率与 OH ,MDA呈正相关 ,与SOD活性呈负相关 (P <0 .0 1 )。结论 :EGCG可能通过减轻 OH对SOD的活性抑制 。
Objective To determine the effect of L epigallocatechins gallic acid ester ( L EGCG) on glomerular mesangial cells (GMCs) proliferation , and to make clear the possible mechanism. Methods We investigated the effect of different doses of L EGCG on GMCs proliferation at 24 h, 48 h and 72 h respectively, and screened the best dosage and time point. GMCs were divided into 5 groups: control, L EGCG group, LPS group, dexamethasone group, and L EGCG plus dexamethasone group. At the end of the experiments, the cultured GMCs and its suspension were collected , the positive rate of proliferating cell nuclear antigen( PCNA) was taken count of by immunohistochemical assay, the rate of effective apoptosis of GMCs by terminal deoxynucleotidyl transferase mediated dUTP nick endlabeling (TUNEL) assay, and OH, malonydialdehyde (MDA), superoxide dismutase (SOD) by biochemical assay. Results L EGCG of 200 mg/L had the most strong inhibitory effect on GMCs at 48 h. The levels of OH,and MDA in the L EGCG group was significantly lower than those in the LPS group ( P <0.01), while the level of SOD in the L EGCG group was significantly higher than that in the LPS group ( P <0.01); the PCNA positive rate of GMCs in the L EGCG group was significantly lower than that in the LPS group ( P <0.01), and the effective apoptosis of GMCs in the L EGCG group were significantly higher than that in the LPS group ( P <0.01). The PCNA positive rate of GMCs was positively correlated with the level of OH and MDA, while negatively correlated with the level of SOD.Conclusion The suppressive effect of L EGCG on the abnormal proliferation of GMCs might be that L EGCG could weaken the suppressive effect of OH on the SOD activity, decrease the accumulation of MDA, and increase the apoptosis of GMCs.
出处
《湖南医科大学学报》
CSCD
北大核心
2003年第3期281-284,共4页
Bulletin of Hunan Medical University
基金
湖南省科委资助项目 (项目编号 :OOSSY3 0 2 2 )
湖南省教育厅资助项目 (项目编号 :0 2A0 15 )
关键词
L-EGCG
脂多糖
诱导
大鼠
系膜细胞增生
细胞凋亡
L epigallocatechins gallic acid ester
glomerular mesangial cells
apoptosis
proliferating cell nuclear antigen
