Effect Of TGF-β₁ On Thyroid Function In Primary Porcine Thyrocytes

ObjectiveTo observe the effect of TGF-β₁ on thyroid function in primary porcine thyrocytes.To approach its mechanism.MethodsTGF-β₁ injured model was made with purified cultured Porcine Thyrocytes ,and the experiment was divided according to the final concentration of TGF-β₁ into 0(control group),2,5,10μg/L. The injured time of TGF-β₁ was 72h. Treated with different dose of TGF-β₁, the cells were observed with a convert microscope in shape, the apoptosis was evaluated by Hoechst 33258 staining techniques.and the cell survival ratio was measured with MTT method and the TPO activity was measured with upgrade guaiacol method. The TPO mRNA expression levels were determined by RT-PCR. To detect T3,T4 by chemiluminescence method .ResultsThyroid cells in primary culture results were observed: After 48h of incubation, the thyroid cell adhesion, which are mostly round or hexagonal, the cell growth showed the trend of aggregation, and can see the typical follicular structure. Add 2,5,10μg / L TGF-β₁ after 72h of culture cells after staining with Hoechst 33258 staining was observed under fluorescence microscope can be seen in the nucleus appears dense concentration of thyroid blue dye staining, showing crescent, round, And the budding and foaming phenomenon can be seen, apoptotic bodies ,floating cells and cell debris can be seen, too.The effect of TGF-β₁ on primary porcine thyroid cell viability: Compared with the control group, 2μg / L, 5μg / L, 10μg / L group were significantly decreased cell survival, the difference was statistically significant (P <0.05);TGF-β₁ on TPO activity: Compared with control group, 2μg / L, 5μg / L, 10μg / L group after TGF-β₁ transfection, TPO activity was significantly decreased, the difference was statistically significant, showed a significant negative correlation (r =- 0.979, P <0.05) ; RT-PCR Results: Compared with the control group, the expression of TPO mRNA has a obviously lower, the difference was statistically significant (P <0.05), and dose - effect relationship.T₃,T₄ levels: With the added concentration of TGF-β₁ was increased, level of T₃,T₄ significantly reduced, the difference between groups was statistically significant (P <0.05);ConclusionsTGF-β₁ on thyroid cells in primary culture had the inhibitory effect,and dose-response relationship,which may be reduced with TPO gene expression in thyroid cells and inhibition of TPO activity and induce cell apoptosis.