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肾脏病与透析肾移植杂志 ›› 2011, Vol. 20 ›› Issue (3): 247-252.

• 论文 • 上一篇    下一篇

氟伐他汀改善嘌呤霉素作用下足细胞β1整合素的表达

  

  • 出版日期:2011-06-30 发布日期:2011-08-08

Fluvastatin on expression of β1 integrin in PAN-treated podocytes

  • Online:2011-06-30 Published:2011-08-08

摘要:

目的:探讨氟伐他汀对嘌呤霉素刺激下足细胞黏附分子β1整合素表达的影响及可能机制。 方法: 体外培养的永生化人足细胞分为以下4组:①正常对照组;②DMSO(1mg/ml)对照组;③嘌呤霉素(PAN, 1.0ug/ml)组:PAN分别培养足细胞12、24、48、72h;④PAN(1.0ug/ml) + 氟伐他汀(fluvastatin,FLV,1×10-8~1×10-5 M)组,嘌呤霉素分别与不同浓度氟伐他汀共同培养足细胞48h。用Western 印迹和DCFHDA(2’,7’-二氯荧光素-3’,6’-二乙酸酯 )分别检测以上4组足细胞β1整合素和活性氧簇(ROS,reactive oxygen species)的表达。MTT(3-(4,5-二甲基-2-噻唑)-2,5-二苯基溴化四唑,简称噻唑蓝)法检测足细胞活力,其分组为:①正常对照组;②DMSO对照组;③PAN(1.0ug/ml)组;④PAN(1.0ug/ml)+ FLV(1×10-8~1×10-5 M)组;⑤FLV(1×10-8~1×10-5 M)组,以上5组分别培养12、24、48、72h。  结果: 与正常对照组相比,PAN分别作用24、48、72h后,β1整合素表达逐渐下降、ROS表达则明显升高(P<0.05)。氟伐他汀干预后,β1整合素表达水平升高、ROS表达水平下降,以氟伐他汀浓度为1×10-7M作用48h效果最明显(P<0.05)。相关性分析发现,PAN组以及PAN+FLV组足细胞β1整合素水平与细胞内ROS表达水平成负相关。MTT结果表明DMSO以及氟伐他汀浓度为1×10-8M和1×10-7M对足细胞的存活无明显影响(P>0.05),而氟伐他汀浓度为1×10-6M和1×10-5M、PAN组、PAN+FLV组,在24h均明显抑制足细胞的活力(P<0.05)。与PAN组相比,嘌呤霉素与1×10-7M氟伐他汀共同作用48h足细胞活力有明显的改善(P<0.05)。 结论: 嘌呤霉素可致体外培养足细胞的β1整合素表达下降,氟伐他汀上调β1整合素的表达,保护足细胞。此机制与氟伐他汀抑制足细胞内ROS的表达相关。

关键词: 氟伐他汀,足细胞,嘌呤霉素 , 活性氧簇

Abstract:

Objective:To investigate the effects and possible mechanism of fluvastatin(FLV) on expression of β1 integrin in PAN-treated podocytes.  Methodology:Cultured human podocytes were divided into 4 groups:They were ①normal control group; ②DMSO(1mg/ml) control group; ③PAN (1.0ug/ml) group:PAN treated podocytes for 12h, 24h, 48h, and 72h respectively; and ④PAN(1.0ug/ml) +FLV(1×10-8~1×10-5 M) group: podocytes were treated with PAN plus different concentrations of FLV for 48h. The expression of β1 integrin and reactive oxygen species (ROS) in all groups was detected by Western Bolt and DCFHDA (2’7’-Dichlorofluoresecein 3’6’-diacetate) respectively. Podocytes were also divided into 5 groups to determine the cellular viability by MTT colorimetry at 12h, 24h, 48h, and 72h respectively.  Results:Compared with the normal control group, the expression of β1 integrin decreased significantly in podocytes treated with PAN at the time of 24h, 48h, and 72h(P<0.05), while the synthesis of ROS increased definitely(P<0.05). Fluvastatin significantly upregulated the expression of β1 integrin and down regulated ROS at the concentration of 1×10-7 M(P<0.05).The expression of β1 integrin had an inverse correlation with the synthesis of ROS in PAN group and PAN+FLV group. The results of MTT revealed that DMSO and lower concentration of fluvastatin (1×10-8M and 1×10-7M) had no significant effect on viability of podocyte(P>0.05), while lower podocyte viability was found in higher concentration of fluvastatin (1×10-6M and 1×10-5M) group, PAN group and PAN+FLV group at 24h(P<0.05). Compared with the PAN group, the viability of podocyte was improved significantly in PAN+FLV group when the concentration of fluvastatin was 1×10-7 M(P<0.05)at the time of 48h. Conclusion:The expression of β1 integrin was decreased in PAN-treated podocytes, which was improved by treating with fluvastatin . The mechanism may be associated with inhibiting the activity of ROS.