Villából kórházat? A József Attiláról elnevezett kórház története

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Inhibitory effect of pirfenidone on TGF-β2-induced fibroblastic phenotypes in SRA01/04 cells.

<p>There was no apparent morphological change in SRA01/04 cells after 24 hours’ treatment with 0.5 mg/ml PFD only (B) compared with control cells(A). In the presence of 12.5 ng/ml TGF-β2, the TGF-β2-induced morphological changes in SRA01/04 cells (C) were detected, including elongated and spindle-like shapes, which were noticeably suppressed by co-treatment with 0.5 mg/ml PFD (D).</p

Cell viability of SRA01/04 cells after treated with pirfenidone for 24 hours in a trypan blue exclusion test.

<p>After 24 hours’ treatment with PFD (0, 0.3, 0.5, and 1 mg/ml), the percentages of living cells were more than 90% in all groups. No statistically significant difference was found between PFD treated and control groups (<i>P</i>>0.05). Data are the mean ± SD of triplicates from an experiment that was repeated with similar results.</p

Inhibitory effect of pirfenidone on TGF-β2-induced migration of SRA01/04 cells.

<p>Light microscope images show decreased migratory ability of cells at 24 hours, after scratches were applied to the cells with 0, 0.25, or 0.5 mg/mL pirfenidone in the absence or presence of TGF-β2. *P<0.05 and **P<0.01 versus the corresponding value for control cells. Data in each bar are the mean ± SD of cells that migrated through the membrane in three separate experiments. Magnification, ×40.</p

Cytotoxicity of pirfenidone in a LDH assay.

<p>After 24 or 48 hours’ treatment with PFD (0, 0.25, and 0.5 mg/ml), no statistically significant difference was found between the percentage of cell-mediated lysis in PFD treated groups and control group (<i>P</i>>0.05). Data are the mean ± SD of triplicates from an experiment that was repeated with similar results.</p

Inhibitory effect of pirfenidone on TGF-β2-induced epithlial-mesenchymal transition in SRA01/04 cells.

<p>Compared with control cells (A), immunocytofluorescence demonstrated that the TGF-beta2 significantly up-regulated the mesenchymal phenotypic marker fibronectin (FN, green) in SRA01/04 cells (C). Either in the absence (B) or presence (D) of TGF-beta2, FN was significantly down-regulated after 24 hours’ treatment with 0.5 mg/ml PFD. The nuclei stained by DAPI (blue). Magnification, ×400.</p

Human Primer Sequences Used for Realtime RT-PCR.

<p>Human Primer Sequences Used for Realtime RT-PCR.</p

Inhibitory effect of pirfenidone on expression of TGFβ2 and SMADs mRNA in SRA01/04 cells.

<p>Real-time RT-PCR showed that pirfenidone reduced the levels of TGFβ2, SMAD3, and SMAD4 when compared with control cells. **P<0.01 versus the corresponding value for control cells. Data are the mean ± SD of triplicates from an experiment that was repeated with similar results.</p

Inhibitory effect of pirfenidone on expression of TGFβ2 and SMADs protein in SRA01/04 cells.

<p>Western blot showed that pirfenidone reduced the protein levels of TGFβ2, SMAD3 and SMAD4 when compared with control cells. *P<0.05 and **P<0.01 versus the corresponding value for control cells. Data are mean ± SD of results from three independent cultures.</p

Expression of TGFβ2 and SMADs protein in SRA01/04 cells after treated with pirfenidone by immunocytochemical detection under confocal microscopy.

<p>TGFβ2 (A), SMAD3 (B), and SMAD4 (C) in the cytoplasm (red) and nuclei (blue) of HLECs were stained by immunocytochemistry after treatment with 0.25 (A2, B2, C2) and 0.5 mg/mL (A3, B3, C3) pirfenidone, respectively. (A0, B0, C0) Corresponding negative controls. Magnification, ×400.</p