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目的研究肝细胞生长因子(HGF)对60Coγ射线照射体外培养的大鼠心肌细胞蛋白表达的影响。方法体外培养新生大鼠心肌细胞,分为未照射组、单纯照射组、HGF处理照射组,照射组细胞分别用60Coγ射线20Gy单剂量照射心肌细胞,HGF处理的照射组在照射前3h用终浓度为40ng/ml的HGF孵育。在照射后48h:(1)用蛋白检测试剂盒检测各组心肌细胞中的总蛋白含量;(2)流式细胞仪检测各组的细胞周期变化;(3)带绿色荧光蛋白基因的腺病毒(AdGFP)感染各组心肌细胞,感染后48h用流式细胞仪检测各组心肌细胞中所含的绿色荧光蛋白的荧光强度,间接比较各组心肌细胞中绿色荧光蛋白的生成量。结果照射后48h,照射组心肌细胞中总蛋白含量较未照射组减低(P<0.01),HGF处理的照射组心肌细胞中总蛋白含量较单纯照射组增高(P<0.05)。照射后96h、感染AdGFP后48h,照射组心肌细胞的绿色荧光强度明显弱于未照射组(P<0.01),HGF处理的照射组心肌细胞的绿色荧光强度较单纯照射组增强(P<0.01)。结论60Coγ射线单剂量照射抑制体外培养的心肌细胞蛋白合成,HGF可部分逆转60Coγ射线对心肌细胞蛋白生成的抑制作用。
Objective To investigate the effect of hepatocyte growth factor (HGF) on the protein expression of cardiomyocytes cultured in vitro by 60Co γ ray irradiation. Methods The neonatal rat cardiomyocytes were cultured in vitro and divided into three groups: unirradiated group, simple irradiation group and HGF-treated irradiation group. The irradiated cells were irradiated with single dose of 20Gy 60Coγ ray, respectively. The HGF- Incubate for 40ng / ml HGF. 48h after irradiation: (1) The total protein content in each group of cardiomyocytes was detected by protein detection kit; (2) the cell cycle changes of each group were detected by flow cytometry; (3) the adenovirus with green fluorescent protein (AdGFP). The fluorescence intensity of green fluorescent protein (GFP) contained in cardiomyocytes of each group was detected by flow cytometry at 48 h after infection, and the amount of green fluorescent protein in each group was indirectly compared. Results 48h after irradiation, the total protein content of myocardial cells in irradiation group was lower than that in non-irradiation group (P <0.01). The total protein content in HGF-treated irradiation group was higher than that of irradiation group (P <0.05). At 96h after irradiation, the green fluorescence intensity of myocardial cells in irradiated group was significantly weaker than that in non-irradiated group (P <0.01) 48h after infection with AdGFP. The green fluorescence intensity of HGF-treated irradiation group was higher than that of pure irradiation group (P <0.01) . Conclusion Single dose of 60Coγ ray can inhibit cardiomyocyte protein synthesis in vitro. HGF can partially reverse the inhibitory effect of 60Coγ-ray on cardiomyocyte protein production.