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Silica promotes the transdifferentiation of rat circulating fibrocytes in vitro.

Yao W, Liu S, Li J, Hao C - Mol Med Rep (2015)

Bottom Line: The mRNA expression levels of collagen I, collagen III and α‑smooth muscle actin (SMA) in cFbs were analyzed by reverse transcription‑quantitative polymerase chain reaction.The results indicated that in the cell model, the free silica effectively increased the protein and mRNA expression levels of collagen‑I, collagen‑III and α‑SMA.The free silica significantly promoted the transdifferentiation of cFbs into myofibroblasts in a dose‑and time-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Department of Public Health, Zhengzhou University, Zhengzhou, Henan 450001, P.R. China.

ABSTRACT
To investigate the effects of silica on circulating fibrocytes (cFbs), the present study established a primary culture model of rat alveolar macrophages and cFbs in vitro. Macrophages were treated with free silica, and their supernatant was used to stimulate cFbs. The mRNA expression levels of collagen I, collagen III and α‑smooth muscle actin (SMA) in cFbs were analyzed by reverse transcription‑quantitative polymerase chain reaction. The intracellular and extracellular protein expression levels of collagen I, collagen III and α‑SMA were detected by ELISA and immunofluorescence staining. The results indicated that in the cell model, the free silica effectively increased the protein and mRNA expression levels of collagen‑I, collagen‑III and α‑SMA. The free silica significantly promoted the transdifferentiation of cFbs into myofibroblasts in a dose‑and time-dependent manner.

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(A and B) Expression levels of collagen I, collagen III and α-SMA in cFbs treated with the supernatant of alveolar macrophages treated with various concentrations of SiO2 for 24 h, as determined by immunofluorescence (magnification, ×100). (C and D) The staining intensity of collagen I, collagen III and α-SMA was quantified by densitometric analysis. Values are expressed as the mean ± standard deviation of three independent experiments (n=6). *P<0.05, vs. the control group. cFbs, circulating fibrocytes; α-SMA, α smooth muscle actin; SiO2, crystalline silica.
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f3-mmr-12-04-5828: (A and B) Expression levels of collagen I, collagen III and α-SMA in cFbs treated with the supernatant of alveolar macrophages treated with various concentrations of SiO2 for 24 h, as determined by immunofluorescence (magnification, ×100). (C and D) The staining intensity of collagen I, collagen III and α-SMA was quantified by densitometric analysis. Values are expressed as the mean ± standard deviation of three independent experiments (n=6). *P<0.05, vs. the control group. cFbs, circulating fibrocytes; α-SMA, α smooth muscle actin; SiO2, crystalline silica.

Mentions: The present study investigated the effects of SiO2 on collagen I, collagen III and α-SMA protein expression at various concentrations and incubation times by immunohistochemistry. As shown in Fig. 3A, collagen I, collagen III and α-SMA was significantly increased in the cFbs following treatment with the supernatant of AM treated with SiO2 (0, 20, 40, 60 and 80 µg/ml) for 24 h. A dose-dependent increase in the expression levels of collagen I, collagen III and α-SMA was observed in the cFbs treated with the supernatant of AM treated with 20, 40 and 60 µg/ml SiO2, as compared with that in the control group (n=6; P<0.05), whereas in the 80 µg/ml SiO2 group, the expression of the respective proteins was similar to that in the 60 µg/ml SiO2 group (P>0.05; Fig. 3C). Fig. 3B shows representative staining for collagen I, collagen III and α-SMA in cFbs following treatment for various durations with the supernatant of AM treated with 80 µg/ml SiO2. The protein expression levels of collagen I, collagen III began to increase on day 3, and those of α-SMA on day 13. As compared with the control group, the expression levels of collagen I and collagen III were significantly upregulated on days 6, 9, 12, 15 and 18 (P<0.05), whereas the expression levels of α-SMA were significantly upregulated on days 16, 19, 22, 25 and 28 (P<0.05; Fig. 3D). These results suggested that SiO2 stimulated the expression levels of collagen I, collagen III and α-SMA in a dose- and time-dependent manner.


Silica promotes the transdifferentiation of rat circulating fibrocytes in vitro.

Yao W, Liu S, Li J, Hao C - Mol Med Rep (2015)

(A and B) Expression levels of collagen I, collagen III and α-SMA in cFbs treated with the supernatant of alveolar macrophages treated with various concentrations of SiO2 for 24 h, as determined by immunofluorescence (magnification, ×100). (C and D) The staining intensity of collagen I, collagen III and α-SMA was quantified by densitometric analysis. Values are expressed as the mean ± standard deviation of three independent experiments (n=6). *P<0.05, vs. the control group. cFbs, circulating fibrocytes; α-SMA, α smooth muscle actin; SiO2, crystalline silica.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581811&req=5

f3-mmr-12-04-5828: (A and B) Expression levels of collagen I, collagen III and α-SMA in cFbs treated with the supernatant of alveolar macrophages treated with various concentrations of SiO2 for 24 h, as determined by immunofluorescence (magnification, ×100). (C and D) The staining intensity of collagen I, collagen III and α-SMA was quantified by densitometric analysis. Values are expressed as the mean ± standard deviation of three independent experiments (n=6). *P<0.05, vs. the control group. cFbs, circulating fibrocytes; α-SMA, α smooth muscle actin; SiO2, crystalline silica.
Mentions: The present study investigated the effects of SiO2 on collagen I, collagen III and α-SMA protein expression at various concentrations and incubation times by immunohistochemistry. As shown in Fig. 3A, collagen I, collagen III and α-SMA was significantly increased in the cFbs following treatment with the supernatant of AM treated with SiO2 (0, 20, 40, 60 and 80 µg/ml) for 24 h. A dose-dependent increase in the expression levels of collagen I, collagen III and α-SMA was observed in the cFbs treated with the supernatant of AM treated with 20, 40 and 60 µg/ml SiO2, as compared with that in the control group (n=6; P<0.05), whereas in the 80 µg/ml SiO2 group, the expression of the respective proteins was similar to that in the 60 µg/ml SiO2 group (P>0.05; Fig. 3C). Fig. 3B shows representative staining for collagen I, collagen III and α-SMA in cFbs following treatment for various durations with the supernatant of AM treated with 80 µg/ml SiO2. The protein expression levels of collagen I, collagen III began to increase on day 3, and those of α-SMA on day 13. As compared with the control group, the expression levels of collagen I and collagen III were significantly upregulated on days 6, 9, 12, 15 and 18 (P<0.05), whereas the expression levels of α-SMA were significantly upregulated on days 16, 19, 22, 25 and 28 (P<0.05; Fig. 3D). These results suggested that SiO2 stimulated the expression levels of collagen I, collagen III and α-SMA in a dose- and time-dependent manner.

Bottom Line: The mRNA expression levels of collagen I, collagen III and α‑smooth muscle actin (SMA) in cFbs were analyzed by reverse transcription‑quantitative polymerase chain reaction.The results indicated that in the cell model, the free silica effectively increased the protein and mRNA expression levels of collagen‑I, collagen‑III and α‑SMA.The free silica significantly promoted the transdifferentiation of cFbs into myofibroblasts in a dose‑and time-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Department of Public Health, Zhengzhou University, Zhengzhou, Henan 450001, P.R. China.

ABSTRACT
To investigate the effects of silica on circulating fibrocytes (cFbs), the present study established a primary culture model of rat alveolar macrophages and cFbs in vitro. Macrophages were treated with free silica, and their supernatant was used to stimulate cFbs. The mRNA expression levels of collagen I, collagen III and α‑smooth muscle actin (SMA) in cFbs were analyzed by reverse transcription‑quantitative polymerase chain reaction. The intracellular and extracellular protein expression levels of collagen I, collagen III and α‑SMA were detected by ELISA and immunofluorescence staining. The results indicated that in the cell model, the free silica effectively increased the protein and mRNA expression levels of collagen‑I, collagen‑III and α‑SMA. The free silica significantly promoted the transdifferentiation of cFbs into myofibroblasts in a dose‑and time-dependent manner.

Show MeSH
Related in: MedlinePlus