复方茵丹汤对HepG2细胞NF-κB、ICAM-1、Egr-1基因沉默后MRP2和RXRα表达的影响

Effect of Compound Yindan Decoction on the expression MRP2 and RXR after silencing of NF-κB、ICAM-1 and EGR-1 genes in siRNA-mediated HepG2 cells

目的:观察HepG2细胞中核因子κB(NF-κB)、细胞间黏附分子-1(ICAM-1)、早期生长反应因子(Egr-1)基因沉默后复方茵丹汤对多药耐药相关蛋白2(MRP2)和人维甲酸X受体α(RXRα)表达的影响。方法:运用RT-PCR和Western blot检测复方茵丹汤对HepG2细胞中NF-κB、ICAM-1、Egr-1沉默与否,对NF-κB、ICAM-1、Egr-1、MRP2和RXRα基因和蛋白水平的影响。结果:与空白对照组比较,复方茵丹汤组和UDCA组NF-κB、ICAM-1、Egr-1基因和蛋白水平明显降低(P<0.01,P<0.05),MRP2和RXRα基因和蛋白水平明显升高(P<0.01,P<0.05),复方茵丹汤对HepG2细胞NF-κB、ICAM-1、Egr-1、MRP2和RXRα表达的诱导作用明显强于UDCA组(P<0.01,P<0.05)。对各组进行NF-κB、ICAM-1、Egr-1基因沉默后,与正常培养的HepG2细胞比较,转染siRNANF-κB、siRNA-ICAM-1和siRNA-Egr-1的细胞,其NF-κB、ICAM-1、Egr-1的基因和蛋白表达水平均显著降低(P<0.01,P<0.05),MRP2和RXRα的基因和蛋白水平显著升高(P<0.01)。与siRNA-NF-κB、siRNA-ICAM-1和siRNA-Egr-1转染组比较,siRNA-NF-κB、siRNA-ICAM-1和siRNA-Egr-1转染+复方茵丹汤组的NF-κB、ICAM-1、Egr-1基因和蛋白的表达水平均显著降低(P<0.01,P<0.05),MRP2和RXRα的基因和蛋白水平显著升高(P<0.01)。结论:复方茵丹汤在体外可通过下调NF-κB、ICAM-1、Egr-1基因和蛋白的表达来上调MRP2和RXRα基因和蛋白的水平。

AIM:To observe the effects of Compound Yindan Decoction on multidrug resistance-related protein 2(MRP2)and retinoic acid X receptor alpha(RXRalpha)in HepG2 cells by SiRNA interference with nuclear factorκB(NF-κB),intercellular adhesion molecule-1(ICAM-1)and early growth response factor(Egr-1)gene silencing.Methods:RT-PCR and Western blot were used to detect the effects of Yindan Decoction on the silence of NF-κB,ICAM-1 and Egr-1 in HepG2 cells,the gene and protein levels of NF-κB,ICAM-1,Egr-1,MRP2 and RXRalpha.Results:Compared with the blank control group,the levels of NF-κB,ICAM-1,Egr-1 gene and protein in Yindan Decoction group and UDCA group decreased significantly(P<0.01,P<0.05),while the levels of MRP2 and RXRalpha gene and protein increased significantly(P<0.01,P<0.05).Yindan Decoction induced the expressions of NF-κB,ICAM-1,Egr-1,MRP2 and RXRalpha in HepG2 cells.The use of UDCA group was significantly stronger than that of UDCA group(P<0.01,P<0.05).After silencing of NF-κB,ICAM-1 and Egr-1 genes in each group,compared with HepG2 cells in normal culture,the gene and protein levels of siRNA-NF-κB,siRNA-ICAM-1 and siRNA-Egr-1 cells transfected with siRNA-NF-κB,siRNA-ICAM-1 and siRNA-Egr-1 decreased significantly(P<0.01,P<0.05),while the gene and protein levels of MRP2 and RXRalpha increased significantly(P<0.01,P<0.05).Compared with siRNA-NF-κB,siRNA-ICAM-1 and siRNA-Egr-1 transfection groups,the expression levels of NF-κB,ICAM-1 and Egr-1 genes and proteins in siRNA-NF-κB,siRNA-ICAM-1 and siRNA-Egr-1 transfection plus compound Yindan Decoction group were significantly decreased(P<0.01,P<0.05),while the gene and protein levels of MRP2 and RXRalpha were significantly increased(P<0.01).Conclusion:Compound Yindan Decoction can up-regulate the levels of MRP2 and RXRalpha genes and proteins by down-regulating the expression of NF-κB,ICAM-1,Egr-1 genes and proteins in vitro.