氟伐他汀改善嘌呤霉素作用下足细胞β1整合素的表达

Fluvastatin on expression of β1 integrin in PAN-treated podocytes

目的:探讨氟伐他汀(FLV)对嘌呤霉素(PAN)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四组:(1)正常对照组;(2)二甲基亚砜(DMSO,1mg/ml)对照组;(3)PAN(1.0μg/ml)组:PAN分别培养足细胞12h、24h、48h、72h;(4)PAN(1.0μg/ml)+FLV(1×10-8～1×10-5M)组,PAN分别与不同浓度FLV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)分别检测以上四组足细胞β1整合素和活性氧簇(reactive oxygen species,ROS)的表达。噻唑蓝[3-(4,5-二甲基-2-噻唑)-2,5-二苯基溴化四唑,简称MTI]法检测足细胞活力。结果:与正常对照组相比,PAN分别作用24h、48h、72h后,β1整合素表达逐渐下降、ROS表达则明显升高(P<0.05)。FLV干预后,β1整合素表达水平升高、ROS表达水平下降,以FLV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FLV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FLV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,PAN与FLV1×10-7M共同作用48h足细胞活力有明显的改善(P<0.05)。结论:PAN可致体外培养足细胞的β1整合素表达下降,FLV上调β1整合素的表达,保护足细胞。此机制与FLV抑制足细胞内ROS的表达相关。

Objective：To investigate the effects and possible mechanism of fluvastatin（FLV） on expression of β1 integrin in PAN - treated podoeytes. Methodology： Cultured human podocytes were divided into 4 groups ： They were （ 1 ） normal control group; （2）DMSO（I mg/ml） control group; （3）PAN （1.0 p,g/ml） group：PAN treated podocytes for 12h, 24h, 48h, and 72h respectively; and （4）PAN（1.0 μg/ml） ＋ FLV（1 ×10^-8-1 ×10^-5 M） group： podoeytes were treated with PAN plus different concentrations of FLV for 48h. The expression of 131 integrin and reactive oxygen species （ROS） in all groups was detected by Western Bolt and DCFHDA （27＇-Dichlorofluoresecein 3＇6＇-diacetate） respectively. Podocytes were also divided into 5 groups to determine the cellular viability by MTF eolorimetry 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.