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. 2023 Jun 10;299(7):104909. doi: 10.1016/j.jbc.2023.104909

Figure 6.

Figure 6

Bioconversion of ALA into DHA inhibits HSC activation by reducing TGFBR1 mRNA expression.A, ALA is biotransformed to DHA by desaturase in the endoplasmic reticulum. B and C, ACTA2, COL1A1 mRNA, and protein levels in LX2 cells after cells were pretreated with DMSO or ALA (0, 5, 10, 20 μM) for 6 h, then stimulated with TGF-β1 (5 ng/ml) for 24 h. D and E, ACTA2, COL1A1 mRNA, and protein levels in LX2 cells after cells were pretreated with DMSO or DHA (0, 5, 10, 20 μM) for 6 h, then stimulated with TGF-β1 (5 ng/ml) for 24 h. F, immunoblotting for SMAD2/3 phosphorylation(p-Smad2/3) levels in LX-2 cells after pretreatment with DMSO, ALA, LA, and DHA for 6 h, then stimulated with TGF-β1 (5 ng/ml) for 30 min. G, TGFBR1, COL1A1 mRNA levels in LX2 cells transfected with siNC or siFADS2 treated with folic acid–free medium (FA-), ALA (10 μM), DHA (10 μM) for 24 h. H, TGFBR1, COL1A1 mRNA levels in LX2 cells transfected with siNC or siFADS2 treated with SHIN1 (10 μM) for 24 h. Phosphorylation of Smad2/3 levels in LX2 cells transfected with siNC or siFADS2 treated with FA-, ALA (10 μM), DHA (10 μM) for 24 h, then stimulated with TGF-β1 (5 ng/ml) for 30 min. (J) is a quantitative analysis of (I). The statistical significance (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001) was tested by student’s unpaired t test. ALA, α-linolenic acid; DHA, docosahexaenoic acid; FADS, fatty acid desaturase; HSC, hepatic stellate cell.