Effects Of Copper On CuZn-SOD Activities And Expression Of CuZn-SOD Gene MRNA In Hepatic Cells In Pigs

The experiment was conducted to evaluate the effects of different copper levels on CuZn-SOD activities and expression of CuZn-SOD gene mRNA in hepatic cells in newborn pigs. In the first experiment, hepatic cells were cultured with 10% blood serum media supplemented with 0, 15.6, 31.2, 46.8, 62.4, 78, 93.6umol/L Cu in 24 aperture culture plate . The results were as follows: CuZn-SOD activities in hepatic cells was significantly increased in culture media added to 0,15.6,31.2,46.8, 62.4, 78, 93.6nmol/L Cu and were highest at 62.4umol/Lcu(P<0.05).In the second experiment, to meaure the expression of CuZn-SOD gene mRNA in hepatic cells, the quantitative reverse transcription-polymerase chain reaction(RT-PCR) was constructed. Hepatic cells was cultured with 10% blood serum media supplemented with 0, 31.2, 62.4, 93.6umol/L Cu in 24 aperture culture plate. The total RNA in hepatic cells was extracted by the Trizol way, CuZn-SOD cDNA was amplified,being ligated with pGEM vector, and the expression of the CuZn-SOD gene mRNA was measured with the RT-PCR. The results were as follows: sequence of the cloned CuZn-SOD gene was identical to Genbank. The expression of CuZn-SOD gene mRNA was enhanced in culture media added to 31.2, 62.4, 93.6umol/L Cu. The optimal copper concentration was 62.4umol/L Cu.We can confer from the above results suggests that as the center of the CuZn-SOD activity Cu can not only increase the activities ,but also control the expression of the gene and enhance the transcription,As the results the increase of transcription offer condition for the translation, which is very important to resist oxidation and offer molecule thereunder for the mechanism of resisting-oxidation of Cu.