| Extracted by supercritical carbon dioxide, the germ oil of Hordeum Vulgare L. var nudum Hook, f was analysed the composition and content of the fatty acid and vitamin through the gas chromatography. Five kinds of fatty acid were determined in the oil, and the content of unsaturated fatty acids in the oil was 79.1% (oleic acid 17%, linoleic acid 55%, Hnolenic acid 7.1%); the content of Vitamin E in the oil is 63.7mg/Kg, and Vitamin D is 36.3mg/Kg; at the same time, we discussed the technology in producing soft capsule beverage of the Hordeum Vulgare L. var nudum Hook, /germ oil.The crude extraction from Hordeum Vulgare L. var nudum Hook, /germ were dialyzed for clearing away big molecule. The dialytic peptides were characterized by chromatography on Sephadex G-25, Sephadex G-15. Some active fractions were separated by reversed-phase high-performance liquid chromatography on Supelco DiscoveryTM C18 column and a linear gradient increasing buffer B from 0% to 60% within 40 min (mobile phase A was water, 0.10%(v/v) TFA, and mobile phase B was 0.10%(v/v) TFA in acetonitrile).Amino acid analysis of the active fractions was performed with amino acid analyzer. The amino acid composition of the peptides reveals mainly ten amino acids: Asp, Glu, Gly, Thr, Ser, Ala, lie, Leu, Phe, Lys. We use high-performance liquid chromatography-mass spectrometry method for characterization of peptide, founding that DNA binding peptide's molecular weight was mainly between 350-1300.The peptide fraction were used to cultured HeLa cells, under the experimental conditions adopted, the result showed that the DNA binding peptides (DBP) has a distinctness inhibitory effect on proliferation of HeLa cells and that permeabilization with calcium phosphate of the plasmatic membrane increases this effect. The peptide is probably internalized by the cells and involved in inhibition of cellular proliferation. Cell numbers were monitored by direct counting using a Coulter Counter ZM hemocytometer. And cell density was assessed by methylene blue exclusion.By phosphorylating the CTD of the biggest subunit of RNA polymerase II, the cyclin-dependent kinases 7 is critical to DNA transcription initiation. The expression pattern of CDK.7 during cultured with DNA binding peptide was determined using an RT-PCR approach. The level of CDK7 gene expression was downregulated by DNA binding peptide in our result. To determine the expression pattern of the CDK7 proteins when HeLa were cultured with extracellular DNA binding peptide, we used western blot and immunohistochemical approach. All the results show that the level of CDK7 protein is also downregulated by DNA binding peptide. To further investigate the changing of CDK7 concentration in HeLa cell after culture with DBP, we found that CDK7 concentration in nuclear decrease more seriously than in cytoplasmic through confocal microscope detecting.We determined the fluctuateing of ERK.1/2 protein concentration in the cell by western blot, and total ERK1/2 was decrease expose to DBP. For detecting whether DBP would down-regulate ERK1/2 activity, we detect the phosphorylation of ERK1/2, which normally reflected ERK.1/2 catalytic activity. In parallel with decreased total ERK1/2, the phosphorylation decreased too. Under the same treatment, the NF-kB p65 protein was determined by western blot and immunichemistroy. The results show that NF-kB decreases in HeLa cell exposed to DBP. Through confocal microscope detecting, we also found that NF-kB p65 distribute throughout cytoplasm and nucleus.Synthetic CTD heptapeptide of 3 repeats is the substrates for CDK.7 in vitro, we use it to culture HeLaparallel with DBP. Although RT-PCR detect CDK7 mRNA with little apparent change in concentration when exposure to synthetic heptapeptides, western blot detected CDK7 protein increasing, similar to the immunocytochemistry results. At the same time, we found that our synthetic CTD peptide don't have significantly effect on ERKl/2 and NF-kB p65 signal pathway.
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