FIGURE 1.

Tetrandrine suppresses TGF-β1-induced myofibroblasts differentiation, proliferation, and ECM deposition in primary MLFs. (A) Chemical structure of TET. Its Molecular formation is C₃₈H₄₂N₂O₆. (B) Primary MLFs were incubated with various indicated concentrations of TET for 24 h and subjected to CCK8 assay to assess cell viability. One-way ANOVA with Dunnett’s multiple comparison test: *p < 0.05 comparison to TET = 0 μM group value: *p < 0.05. (C,D) Primary MLFs were pretreated with DMSO or TET (4 μM) for 1 h and stimulated with or without TGF-β1 (10 ng/ml) for 24 h. Representative immunoblot analysis (C) and quantitative analysis (D) show the expression of ECM deposition (fibronectin, COL-I), myofibroblasts transdifferentiation (vimentin, α-SMA). (E,F) Representative Immunofluorescence monitored by confocal microscopy (E) and quantitative analysis (F) show the expression of α-SMA (red) in Primary MLFs. Blue staining indicates nuclei Scale bars: 100 μM. (G,H) Representative Immunofluorescence (G) and quantitative analysis (H) of EdU -positive cell proportion show the proliferation marker (EdU: 5-ehtynal-2′-deoxyuridine) in myofibroblast. Scale bars: 100 μM. (I-K) Representative immunoblot analysis (I) and quantitative analysis (J,K) show the expression of TGF-β/smad pathway markers (p-smad2, smad2, p-smad3, smad3) in primary MLFs. p values were determined by two-way ANOVA with Tukey’s multiple comparison test (n = 5): *p < 0.05.