The Effect Of High Expression Of Mouse Proxisome Proliferator Activated Receptor γ₁ On Free Fatty Acid-Induced β TC3 Cells Impairment

Objective: To clone mouse proxisome proliferator activated receptor γ/ (mPPAR γ/) for expressing stably in pancreatic β TC3 cells . Then to observe the effect of high expression of mPPARγ1 on free fatty acid (FFA)-induced β TC3 cells impairment.Methods: mPPARγ1 cDNA complete sequence was amplified by RT-PCR from the total RNA of epididymis fat pad of Chinese Kunming mice and cloned into plasmid pcDNA3.1 to generate the recombinant plasmid pcDNA3.1/ mPPAR γ1 which was confirmed to contain the amplified target gene segments with sequencing. The recombinant plasmid pcDNA3. 1/ raPPAR γ1 was used to stably transfect pancreatic β TC3 cells with lipofectamine and the expression was detected with semi-quatitive RT-PCR. Then the cell viability of wild β TC3 cells was compared with that of the β TC3 cells high expressing mPPAR γ1 with MTT viability assay after they were exposed long to high-level FFA.Results: The sequencing results for the amplified target gene showed that the sequence of mPPAR γ1 gene from the epididymis fat pad of Chinese Kunming mice is similar to that of mouse PPAR γ1 gene in Genbank, only the codon coding Asn at the site of 421 amino acid changed from AAU to AAC. mPPAR γ1 efficiently expressed in β TC3 cells in vitro. The cell viability of wild βTC3 cells reduced after exposed to high-level FFA for long time. Higher the level of FFA was, more obvious the reductionof the cell viability was. However, at the same condition, the cell viability of the β TC3 cells high expressing mPPARγ1 has no distinct change.Conclusion: The high expression of PPAR γ1 could protect βTC3 cells from FFA-induced impairment.