Effect of Ca2＋-Calcineurin on fibroblast-to-myofibroblast differentiation

Objective To explore the effect of Ca2＋-Calcineurin on fibroblast-to-myofibroblast differentiation. Methods The lung fibroblasts were isolated from the lungs of SD rats at postnatal day 3. Use immunohistochemical technique to observe the expression of Vimentin. The fluorescence intensity of intracellular Ca2＋ was detected respectively with Fluo-3/AM loading by the laser confocal microscopy. The expression of α-SMA and p-Smad2 were measured by Western blotting. Results ①It can observe the expression of vimentin in primary lung fibroblasts. （2）TGF-β1 can increase of cytosolic Ca2＋ of primary lung fibroblasts.③TGF-β1 stimulated group was obvious increased the expression of α-SMA compared to Control group,CsA as the inhibitor of calcineurin can＇t inhibit the expression of α- SMA. TGF-β1 together with CsA stimulated of 1 h,4 h,24 h group were obvious increased the expression of p-Smad2 compared to TGF-β1 stimulated group,and 1 h group was more than the other groups. There were no differentiation between 4 h group and 24 h group. Conclusions There were changes of cytosolic Ca2＋ in rat lung fibroblasts by TGF-β1. CaN may take part in the process of fibroblast-to-myofibroblast differentiation. Key words: Pulmonary fibrosis; Ca2＋ ; Calcineurin