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目的研究锰对肉鸡心肌细胞MnSOD基因的转录调节。方法将144只1日龄肉公鸡随机分为3组,分别饲喂不添加锰的基础饲粮(对照组)或添加100、200mg/kg锰(试剂级硫酸锰)的饲粮组,饲喂21d。采用全谱直读型ICP发射光谱仪分析心肌锰含量;用RT-PCR技术分析心肌细胞MnSOD基因的mRNA水平;用电泳迁移率变动分析法(EMSA)检测转录因子Sp-1和AP-2的DNA结合活性。结果心肌锰含量(P<0.002)和MnSOD mRNA水平(P<0.0001)随饲粮锰添加水平的升高而显著升高。心肌细胞转录因子Sp-1的DNA结合活性随饲粮锰添加水平的升高显著升高(P<0.0001),而其AP-2的DNA结合活性则随饲粮锰添加水平的升高显著降低(P<0.002)。结论锰可以调节控肉鸡心肌细胞MnSOD基因的转录。
Objective To study the transcriptional regulation of MnSOD gene in broiler cardiomyocytes by manganese. Methods One hundred and fourteen day-old broilers were randomly divided into three groups and fed with the basal diet without manganese supplementation (control group) or with 100,200 mg / kg manganese (reagent grade manganese sulfate) 21d. The content of manganese in myocardium was analyzed by full-spectrum direct-reading ICP emission spectrometer. The mRNA level of MnSOD gene in cardiomyocytes was analyzed by RT-PCR. The DNAs of Sp-1 and AP-2 were detected by electrophoretic mobility shift assay (EMSA) Binding activity. Results Manganese content (P <0.002) and MnSOD mRNA level (P <0.0001) were significantly increased with the increase of dietary Mn levels. The DNA binding activity of cardiomyocyte transcription factor Sp-1 was significantly increased with dietary manganese supplementation (P <0.0001), while DNA-binding activity of AP-2 was significantly decreased with dietary manganese supplementation (P <0.002). Conclusion Manganese can regulate the transcription of MnSOD gene in broiler cardiomyocytes.