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. 2019 Dec 5;14(12):e0225911. doi: 10.1371/journal.pone.0225911

Fig 5. Resveratrol prevents HG-induced collagen type I and DDR2 in vascular adventitial fibroblasts.

Fig 5

(A-F) Sub-confluent quiescent cultures of vascular adventitial fibroblasts in M199 were pre-treated with 50 μM resveratrol for 1 h and subsequently with HG for 12 h. Protein was isolated at 12 h post-HG treatment and subjected to western blot analysis for detection of: (A and B) Collagen type I alpha 1; *p< 0.05 vs. control, ## p< 0.01 vs. HG (One-way ANOVA p< 0.05), and (C and D) DDR2; *p< 0.05 vs. control, ## p< 0.01 vs. HG (One-way ANOVA p< 0.05). (E and F) TGF-β1; ***p< 0.001 vs. control, ### p< 0.001 vs. HG (One-way ANOVA p< 0.05). β-actin was used as loading control. (G and H) Vascular adventitial fibroblasts were transiently transfected with DDR2 overexpression vector. Post-transfection, following revival and serum deprivation, the cells were pre-treated with resveratrol for 1 h, followed by stimulation with HG for 12 h. Collagen type I alpha 1 protein was examined by western blot analysis. ***p< 0.001 vs. control, ### p<0.001 vs. HG, ††† p< 0.001 vs. HG+RESV. (I and J) Vascular adventitial fibroblasts were pre-treated with resveratrol for 1 h, followed by stimulation with HG and exogenously added recombinant TGF-β1 (10ng/mL). Protein expression levels of collagen type I alpha 1 and DDR2 were examined at 12 h by western blot analysis. ** p<0.01 vs. control, ## p<0.01 vs. HG, †† p<0.01 vs. HG+RESV. Data are representative of three independent experiments, n = 3. Error bars represent SD.