Figure 5. Gene expression in rabbit AFSCs cultured in growth medium with five different conditions for 6 days and tested by qRT-PCR. (A) Collagen I gene expression; (B) MMP-3 gene expression; (C) MMP-13 gene expression; (D) IL-1β gene expression; (E) IL-6 gene expression; (F) TNF-α gene expression. The results indicated that low concentration of bleomycin did not change the expression of collagen I in AF cells, however, high concentration of bleomycin (50 μg/ml) decreased collagen type I expression in AF cells. Adding low concentration of rapamycin (25 nM) in bleomycin treated AF cells didn’t change the collagen I expression, but high concentration of rapamycin (50 nM) increased by 50% the collagen I levels in AF cells (A). In addition, the gene expression of MMP-3 (B) and MMP-13 (C) was increased in a bleomycin concentration-dependent manner. Rapamycin decreased the expression of MMP-3 and MMP-13 in AF cells compared to bleomycin alone. Moreover, inflammatory-related genes, such as IL-1β (D), IL-6 (E) and TNF-α (F) were increased in bleomycin treated AF cells; rapamycin inhibited the expression of these inflammation genes. *p<0.05 compared to control, #P<0.05 compared to bleomycin 50 μg/ml.