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. 2022 Oct 19;13:981817. doi: 10.3389/fphar.2022.981817

FIGURE 1.

FIGURE 1

CBD inhibits collagen transcription and synthesis in vitro without altering SMAD2 pathway. (A) Col1a2 transcriptional activity in NIH-3T3 cells pretreated with CBD for 1 h and stimulated with TGFβ1 for the following 24 h (n = 4) (B) Effect of CBD pretreatment on soluble collagen release by human dermal fibroblast using the Sircol Assay. Fibroblasts were pretreated with CBD for 1 h and stimulated with TGFβ1 for 48 h and collagen was measured in the culture media (n = 3). (C) Effect of CBD on SMAD-dependent transcriptional activity. NIH-3T3 cells were transfected with the CAGA-Luc plasmid, preincubated with the indicated concentrations of CBD for 1 h and stimulated with TGFβ1 (10 ng/ml) for 6 h. Then, cells were lysed for luciferase activity (n = 3). Data are expressed as mean ± S.D. relative to control cells. *p < 0.05; **p < 0.01; ***p < 0.001 vs TGFβ1-treated cells. (D) CBD effect on SMAD2 phosphorylation. NHDF cells were incubated in low serum conditions (1% FBS) for 24 h. Then cells were pretreated with CBD or Rosiglitazone (RGZ) for 1 h and stimulated with TGFβ 1 (10 ng/ml) for 2 h. (E) Viability of NIH3T3 cells treated with different concentrations of CBD after 24 h expressed as percentage taking control as 100% (n = 3). (F) Effect of CBD treatment on Col1a2 transcriptional activity in NIH-3T3 cells (n = 3).