Figure 3 Icariin restored mitochondrial homeostasis in UUO
mice. (A) Representative immunofluorescence staining and quantitative
of TOMM20 showed that icariin increased mitochondrial mass. (B)
Representative transmission electron microscope images of mitochondria
in renal tubular epithelial cells. (C) Quantification of the percentage
of damaged mitochondria. * p < 0.05, ** p < 0.01.
Icariin inhibited profibrotic phenotype and inflammatory
response in TGF-β1-exposed HK-2 cells
To further validate the protective role of icariin in tubulointerstitial
fibrosis, TGF-β1-treated HK-2 cells were used to detect the effects of
icariin in vitro . Firstly, we assessed the cytotoxic effect of
icariin on HK-2 cells and observed that the cytotoxic concentration
exceeded 200μM (Figure 4A). Based on published literatures and
preliminary experiments, we determined that the effective concentration
of icariin was 50µM. Then, we measured whether icariin (50µM) could
inhibit profibrotic effects of TGF-β1 in HK-2 cells. Similar to the
findings in the in vivo model, icariin dramatically inhibited
profibrotic phenotype of TGF-β1-treated HK-2 cells, as demonstrated by
the reduction of collagen I and α-SMA expression levels (Figure 4B).
Additionally, quantitative real-time PCR of IL-1β, TNF-α and IL-6
demonstrated that icariin reduced the mRNA expression of inflammatory
factors
in
HK-2 cells (Figure 4C-E). These results validated the anti-inflammatory
and antifibrosis properties of icariin in vitro .