槲皮素对多柔比星致乳大鼠心肌细胞抗氧化酶活性及Fas/FasL表达变化的影响

目的探讨槲皮素对多柔比星致乳大鼠心肌细胞损伤的保护作用及机制。方法原代培养乳大鼠心肌细胞,随机分为正常对照组、多柔比星1.72μmol·L-1损伤组和多柔比星+槲皮素25,50及100μmol·L-1组。槲皮素与心肌细胞培养3h后,加入多柔比星继续培养24h,正常对照组仅加等量DMEM培养液。倒置显微镜下观察细胞生长状态,比色法检测培养液中谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)的活性,用RT-PCR和Western印迹法检测Fas和FasL的mRNA和蛋白的表达。结果与正常对照组相比,多柔比星损伤组心肌细胞生长状态差,GSH-Px和SOD的活性降低,Fas和FasL的mRNA和蛋白的表达均升高。槲皮素25,50及100μmo·lL-1均可拮抗多柔比星所致的上述变化:GSH-Px分别为(76±3),(73±4),(71±3)vs(69±3)kU·L-1;SOD活性分别为(31±2),(29±2),(29±2)vs(26±2)kU.L-1;Fas mRNA:0.61±0.11,1.04±0.12,1.29±0.11 vs 1.61±0.16;FasL mRNA:0.81±0.07,1.24±0.10,1.57±0.09vs1.79±0.11;Fas蛋白:1.08±0.12,1.54±0.10,1.89±0.11 vs 2.15±0.15;FasL蛋白:1.51±0.08,1.70±0.12,2.20±0.09 vs 2.41±0.26。结论槲皮素可减轻多柔比星致乳大鼠心肌细胞凋亡,Fas和FasL蛋白表达的减少是其可能机制之一。

Effects of quercetin on antioxidase activity and Fas/FasL expression in neonate rat cardiomyocytes injured by doxorubicin

OBJECTIVE To study the protective effect and mechanism of quercetin(Que) on doxorubicin(Dox)-induced damage to cardiomyocytes in neonate rats in terms of Fas/FasL and oxidative stress. METHODS Cultured neonatal rat cardiomyocytes were randomly divided into 5 groups: normal control group, Dox 1.72 μmol·L^-1 group, Dox+Que 25, 50 and 100 μmol·L^-1 groups. The cells in Dox+Que groups were pretreated with Que for 3 h and then co-incubated with Dox for 24 h. To observe the cell growth condition by inverted microscope, the activity of glutathion peroxidase (GSH-Px) and superoxide dismutase (SOD) was detected by chromatometry; the mRNA and protein expression of Fas and FasL was detected by RT-PCR and Western blotting, respectively. RESULTS Compared with normal control group, the activity of GSH-Px and SOD in Dox group decreased; and the mRNA and protein expression of Fas and FasL was up-regulated. Que 25，50 and 100 μmol·L^-1 could act as an antagonist of changes induced by Dox：GSH-Px, (76±3)，(73±4)，(71±3) vs (69±3) kU·L^-1；SOD, (31±2)，(29±2)，(29±2) vs（26±2）kU·L^-1；Fas mRNA：0.61±0.11, 1.04±0.12, 1.29±0.11 vs 1.61±0.16; FasL mRNA：0.81±0.07, 1.24±0.10, 1.57±0.09 vs 1.79±0.11; Fas protein：1.08±0.12, 1.54±0.10, 1.89±0.11 vs 2.15±0.15; FasL protein：1.51±0.08, 1.70±0.12, 2.20±0.09 vs 2.41±0.26. CONCLUSIONQue may significantly inhibit the apoptosis induced by Dox in the cultured myocardial cells. The potential mechanism is that Que can down-regulate Fas and FasL protein expression.