Figure 5.

Angiotensin II induces CTGF production and markers of liver fibrosis. Angiotensin II was used to increase CTGF production and induce fibrosis in liver spheroids. Blue bars present the mRNA level data, red bars, results of IHC analysis, and green bars, data acquired using ELISA. (A) Angiotensin II caused an increase in CTGF expression (n = 4; 3 experiments with the PHH donor 1 and 1 experiment with PHH donor 2; NPC from donor 3 in all 4 experiments) and production (n = 4; 3 experiments with the PHH donor 1 and 1 experiment with PHH donor 2; NPC from donor 3 in all 4 experiments). (B) Expression and protein production of COL1A1 increased after angiotensin II treatment (n = 4). (C) TGF-β1 was increased on the protein level (n = 4), however the transcriptional level was unaffected (n = 4). (D) Pro-collagen I α1 released from the cells increased during the last 2 days of culture of the angiotensin II-treated spheroids (n = 4). (E) Similarly, the CTGF concentration in the culture medium was increased in all 4 experiments, although not to a significant extent (n = 4). (F,G) LOX and IL6 mRNA expression increased due to angiotensin II treatment (n = 4; 3 experiments with PHH donor 1 and 1 experiment with PHH donor 2; NPC from donor 3 in all 4 experiments). Nuclei in the IHC images are shown in blue. Every dot in the plots represents data from 1 experiment. Every dot in the red graphs is the average of at least 10 analyzed sections of spheroids per experiment. The size of the spheroids is marked with a white scale bar of 100 μm. Data are shown as mean ± SEM. Statistical analysis—Student’s t-test; * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant. Ang-II—angiotensin II.