论文部分内容阅读
目的 探讨维生素E(VitaminE ,VitE)对大鼠肾小球系膜细胞 (GMCs)增生的影响及可能机制。方法 应用不同剂量 ( 5 0mg/L、10 0mg/L、2 0 0mg/L)VitE在不同时间 ( 2 4h、48h、72h)对GMCs增生的影响 ;选定最佳剂量与最佳时间 ,将GMCs分为脂多糖 (LPS)组、对照组、VitE组、激素组与激素+VitE组共 5组分瓶培养 ,后三组加LPS诱导实验末 ,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 (PCNA)阳性表达率 ,TUNEL法检测GMCs有效凋亡率 ;应用生化法对培养上清液进行羟自由基 ( OH)、总超氧化物歧化酶 (tSOD)、丙二醛 (MDA)的测定。结果 10 0mg/LVitE组在 48h( 3 3 3± 2 3 )对GMCs增生的抑制效果好于其他浓度与时间组 ;VitE组细胞上清中 OH( 2 0 5±3 8)、MDA( 5 7± 0 6)低于LPS组 ( 2 93± 42 ,19 3± 6 0 ) ,tSOD( 10 4± 3 1)活性高于LPS组 ( 15 6± 2 1) (P均 <0 0 1) ;VitE组培养细胞PCNA阳性率 [( 3 3 3± 8 8) % ]低于脂多糖组 [( 46 8± 5 3 ) % ],有效凋亡率高于脂多糖组 (P均 <0 0 1)。PCNA阳性率与 OH、MDA正相关 (r =0 880 ,0 70 2 ,P均 <0 0 1) ,与tSOD活性负相关 (r=- 0 682 ,P <0 0 1)。结论 VitE可以上调tSOD的活性 ,抑制系膜细胞 OH的分泌与MDA的过度积聚
Objective To investigate the effect of vitamin E (VitE) on the proliferation of rat mesangial cells (GMCs) and its possible mechanism. Methods The effects of different doses (50 mg / L, 100 mg / L, 200 mg / L) of VitE on the proliferation of GMCs at different times (24 hours, 48 hours and 72 hours) were studied. The best dosage and the best time GMCs were divided into five groups: LPS group, control group, VitE group, hormone group and hormone + VitE group. The last three groups were cultured with LPS, and cultured cells and smears were collected. Immunohistochemistry The positive rate of PCNA expression in GMCs was detected by enzyme linked immunosorbent assay (ELISA). The effective rate of apoptosis of GMCs was detected by TUNEL assay. The contents of hydroxyl radical (OH), total superoxide dismutase (tSOD) Determination of dialdehyde (MDA). Results The inhibitory effect of 10 0 mg / LVitE group on GMCs hyperplasia at 48h (3 3 3 ± 2 3) was better than that in other concentrations and time groups. OH (20 05 ± 8 8), MDA ± 10 6) was lower than that in LPS group (93 ± 42, 19 3 ± 60), tSOD (104 ± 3 1) was higher than that in LPS group (P <0.01). The positive rate of PCNA in vitE group was lower than that in lipopolysaccharide group [(463 ± 8 8)%] (46 8 ± 5 3%), the effective apoptosis rate was higher than that in lipopolysaccharide group (P <0.01) ). The positive rate of PCNA was positively correlated with OH and MDA (r = 0 880, 0 70 2, P <0 01), and negatively correlated with tSOD activity (r = - 0 682, P 0 01). Conclusion VitE can up-regulate the activity of tSOD and inhibit the secretion of OH and the excessive accumulation of MDA in mesangial cells