柔红霉素对大鼠心肌细胞脑利钠肽表达影响的研究

Effects of daunorubicin on the expression of brain natriuretic peptide gene in myocardial cells:an in vitro experiment with cultured myocardial cells from rats

目的研究柔红霉素对大鼠心肌细胞脑利钠肽表达的影响。方法体外分离培养心肌细胞,实验分正常对照组、柔红霉素组(柔红霉素0.2 mg/L 加入培养液)及柔红霉素+1、6-二磷酸果糖组(柔红霉素0.2 mg/L 及1、6-二磷酸果糖2 mg/ml 分别加入培养液),用四唑盐比色试验、RT-PCR 及 Western 印迹等方法检测各组细胞增殖及脑利钠肽表达。结果应用柔红霉素后大鼠心肌细胞脑利钠肽 mRNA 表达与内参 GAPDH 表达平均灰度比值从(0.94±0.25)减少到(0.32±0.18,P<0.01),蛋白印迹与内参 GAPDH 平均光度比值从(0.82±0.13)减少到(0.40±0.09,P<0.01),加入1、6-二磷酸果糖干预后其 mRNA 表达与内参 GAPDH 平均灰度比值及蛋白印迹与内参 GAPDH 平均光度比值分别为(0.36±0.22)和(0.42±0.11),均无明显增加。结论柔红霉素对心肌细胞脑利钠肽表达呈抑制作用,1、6-二磷酸果糖不能逆转其抑制作用。

Objective To investigate the effects ofdaunorubicin（DNR） on the gene expression of brain natriuretic peptide（BNP） in myocardial cells. Methods Myocardial cells were isolated from neonatal Wistar rats, cultured, and added with DNR of the concentrations of 0, 0.02, 0.2, and 2 mg/L respectively so as to calculate the maximum non-toxic concentration of DNR. The cultured rat myocardial cells were divided into 3 groups,DNR group, added with DNR 0.2 rag/L, DNR ＋ fructose-1 ,6-diphosphate （FDP） group, added with DNR 0. 2 mg/L and FDP 2 mg/ml, and control group. The viability of the myocardial cells was examined by MTT method. RT-PCR and Western blotting were used to detect the mRNA expression and protein expression of the BNP gene. Results Toxicity test showed that the maximum non-toxic concentration of DNR was 0. 2 mg/L. Western blotting showed that the average grey level ratio of BNP/ GAPDH of the DNR group was 0.40 ± 0.09, not significantly different from that of the DNR ＋ FDP group （0.42±0. 11, but significantly lower than that of the control group （0. 82±0. 13, P 〈0.01 ）. Conclusion DNR suppresses the expression of BNP gene in myocardial cells and FDP cannot reverse the suppressing effect.