A human thioredoxin preparation of gene-modified hepatocytes.docVIP

 PAGE \* MERGEFORMAT 15 A human thioredoxin preparation of gene-modified hepatocytes Of: Li Hua, Wang root tree, Zhang Jian, Jiang Nan, Guchang Chang, Yang Yang, Chen planning [Abstract] [aim] prepared a human thioredoxin (hTrx) gene-modified hepatocytes. [Methods] Reverse transcriptase polymerase chain reaction amplified hTrx cDNA, preparation of recombinant retroviral gene modification hTrx large rat liver cells. albumin immunohistochemistry SABC method of liver cell activity in identification, MTT colorimetric tests ability to detect oxidative stress. [Results] hTrx open reading frame of cloned cDNA and preparation of the recombinant retrovirus infection of eukaryotic cells may secrete the hTrx fusion expression and biological activity. hTrx prepared gene-modified rat liver cells by immunohistochemistry SABC method showed normal function of liver cells, MTT assay for measuring oxidative stress has a strong capacity ( P lt;0.05), and can effectively proliferation. [Conclusion] successfully prepared a strong anti-oxidative stress hTrx genetically modified liver cells. [Keywords:] liver cells; human thioredoxin; genetically modified Abstract: [Objective] To prepare the human thioredoxin (hTrx) gene-modified hepatocytes. [Methods] hTrx cDNA was amplified by reverse transcription-polymerase chain reaction (RT-PCR), recombinant retrovirus was applied to primary cultured rat hepatocyte for infection to generate hTrx gene-modified rat hepatocytes, whose viability and antioxidative capacity were examined with albumin-immunohistochemical staining and MTT assay, respectively. [Results] The hTrx open reading frames cDNA was cloned and assembled with recombinant retrovirus, then eukaryotic cells infected by this virus were capable of expressing bioactive hTrx in the form of fusion proteins. Immunohistochemistry demonstrated normal function of the hTrx gene-modified hepatocytes, which possessed strong antioxidative capacity as shown by MTT assay and could be p