摘要
目的 研究维生素E(VitE)对大鼠肾小球系膜细胞 (GMCs)单核细胞趋化蛋白 1(MCP 1)、一氧化氮 (NO)表达的影响。方法 将研究大鼠分对照组、脂多糖 (LPS)组、5 0mg/LVitE +LPS组、10 0mg/LVitE +LPS组、2 0 0mg/LVitE +LPS组共 5组培养肾小球系膜细胞 ,分 2 4、4 8、72h 3个时间段收集培养上清及培养细胞 ;实验末应用生化法测定培养上清NO水平 ,应用ELISA技术检测培养上清MCP 1水平 ,应用RT PCR检测MCP 1mRNA表达。结果 2 4h末LPS组GMCsNO分泌均较对照组降低 (P <0 .0 1) ,其他各组GMCsNO分泌均较对照组增高 (P均 <0 .0 1) ,MCP 1分泌与表达均较对照组增高 (P均 <0 .0 1) ;4 8h末各组NO分泌均较对照组降低 ,LPS组在 4 8h末与 72h末MCP 1分泌较其他各组明显增高 (P均 <0 .0 1) ;与LPS组相比 ,4 8h末与 72h末不同浓度VitE组MCP 1分泌降低 ,与对照组比较各组在不同时间段MCP 1mRNA表达明显增高 (P均 <0 .0 1) ,MCP 1mRNA表达在 4 8、72h较LPS组明显降低 (P均 <0 .0 1) ,且发现各组MCP 1mRNA表达在4 8、72h无明显差异 ,10 0mg/LVitE组与 2 0 0mg/LVitE组较 5 0mg/LVitE组MCP 1mRNA表达明显降低 (P均 <0 .0 1) ,但两组间无明显差异 ,经相关分析表明 ,NO分泌与MCP 1分泌与表达明显负相关 (γ =0 .6 13、0 .
Objective To study the effect of vitamin E(VitE) on expression of monocyte chemo-attractant protein-1(MCP-1) and nitric oxide (NO) in renal glomerular mesangial cells (GMCs)of rats. Methods These rats were divided into 5 groups: control group, lipo-polysaccharide (LPS) group, VitE (50 mg/L, 100 mg/L,200 mg/L,respectively)+LPS groups. After 24 h, 48 h and 72 h,GMCs culture, the supernatant and the GMCs suspension were collected respectively. The levels of NO (nitric oxide, by biochemical methods) and MCP-1( by ELISA methods) in supernatant were detected, and MCP-1 mRNA expression of GMCs was detected by RT-PCR technique.Results 1.The levels of NO secretion: at the end of 24 h culture, the level of NO secretion in GMCs of LSP group markedly decreased compared with those of control groups (all P<0.01), while those of other 3 VitE+LPS groups markedly increased (all P<0.01); after 48 h culture, the levels of NO secretion markedly decreased in all experimental groups, much lower than controls, especially in LSP group compared with control groups (all P<0.01). 2. Secretion level of MCP-1: ①after 24 h,48 h and 72 h of culture, the secretion of MCP-1 in all experimental groups markedly increased compared with those of the control groups(all P<0.01);②secretion of MCP-1 in all experimental groups reached their peak at the time of 48 h of culture and remained high level over 72 h;③the secretion of MCP-1 at 48 h and 72 h in LSP group was markedly higher than those of other expe-rimental groups (VitE+LSP groups,all P<0.01). 3.Expressions of MCP-1 mRNA: ①compared with control groups, the expressions of MCP-1 mRNA (detected in 24 h,48 h and 72 h respectively) in all experimental groups elevated markedly (all P< 0.01), while 48 h and 72 h the expressions of MCP-1 mRNA in LSP group was higher than other VitE+LSP groups(all P< 0.01);②there were no significant changes of MCP-1 mRNA expressions in all experimental groups at 48 h and 72 h. ③Expressions of MCP-1 mRNA in VitE(50 mg/L)+LSP group was higher than VitE(100 mg/L)+LSP and VitE(200 mg/L)+LPS groups(all P<0.01). Correlation analysis showed that there was negative correlation between NO secretion and MCP-1 expression(γ=0.613, 0.682,respectively P<0.01). Conclusions VitE may have an effect on inhibiting the expression and secretion of MCP-1 by promoting the secretion of NO. On the other hand, VitE may influence MCP-1 expression through the translation of MCP-1 mRNA, lessen immune injury of GMCs, and delay chronic deterioration of glomerular diseases.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2004年第9期732-735,共4页
Journal of Applied Clinical Pediatrics
基金
国家自然科学基金资助项目 (39970 776)
深圳市卫生局科研资助项目 (2 0 0 0 60 0 3)
