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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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mRNA expression levels of collagen I, collagen III and α-SMA in cFbs treated with the supernatant of AM treated with various concentrations of SiO2. The cFbs were treated with the supernatant of AM treated with SiO2 at concentrations of 0, 20, 40, 60 or 80 µg/ml for 24 h, and the control group was not treated. (A) The cFbs were harvested, total RNA was extracted and the mRNA expression levels were determined by reverse transcription-quantitative polymerase chain reaction. (B) The mRNA expression levels of collagen I, collagen III and α-SMA were quantified and normalized relative to internal β-actin mRNA. The bands were quantified by densitometric analysis. Values are expressed as the mean ± standard deviation from three independent experiments (n=6). *P<0.01, vs. the control group. cFbs, circulating fibrocytes; α-SMA, α smooth muscle actin; SiO2, crystalline silica; AM, alveolar macrophages.
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f5-mmr-12-04-5828: mRNA expression levels of collagen I, collagen III and α-SMA in cFbs treated with the supernatant of AM treated with various concentrations of SiO2. The cFbs were treated with the supernatant of AM treated with SiO2 at concentrations of 0, 20, 40, 60 or 80 µg/ml for 24 h, and the control group was not treated. (A) The cFbs were harvested, total RNA was extracted and the mRNA expression levels were determined by reverse transcription-quantitative polymerase chain reaction. (B) The mRNA expression levels of collagen I, collagen III and α-SMA were quantified and normalized relative to internal β-actin mRNA. The bands were quantified by densitometric analysis. Values are expressed as the mean ± standard deviation from three independent experiments (n=6). *P<0.01, vs. the control group. cFbs, circulating fibrocytes; α-SMA, α smooth muscle actin; SiO2, crystalline silica; AM, alveolar macrophages.

Mentions: To evaluate the effects of SiO2 on collagen I, collagen III and α-SMA mRNA expression in cFbs, the cFbs were treated with the supernatant of SiO2-treated AM (20, 40, 60 and 80 µg/ml for 24 h). Treatment with the supernatant of AM treated with various concentrations of SiO2 increased the mRNA expression levels of collagen I, collagen III and α-SMA in the cFbs. This increase in expression levels became apparent at 20 µg/ml SiO2 and markedly apparent at 80 µg/ml SiO2. The expression of collagen I, collagen III and α-SMA was significantly and dose-dependently upregulated following treatment with 20, 40 and 60 µg/ml SiO2, as compared with that in the control group (P<0.05), while no statistically significant difference was observed between the 60 µg/ml and 80 µg/ml groups (P>0.05; Fig. 5A and B). Following treatment of the cFbs with the supernatant of AM treated with 80 µg/ml SiO2 for 28 days, the mRNA expression levels of collagen I, collagen III and α-SMA were determined by RT-qPCR. As shown in Fig. 6, increases in collagen I and collagen III expression were detected on day three, and increases in α-SMA expression was observed on day 13 of incubation, whereas in the control and 0 µg/ml groups, elevated collagen I and collagen III expression was detected on day 6, and elevated α-SMA expression on day 19. A significant upregulation of the expression levels of collagen I and collagen III in the treated groups was observed on days 3, 6, 9, 12, 15 and 18 (P<0.05). The expression levels of α-SMA were significantly upregulated on days 13, 16, 19, 22, 25, 28, as compared with those in the control group (P<0.05). These results suggested that SiO2 was able to stimulate collagen I, collagen III and α-SMA activity at the transcriptional level, and increase the mRNA synthesis of collagen I, collagen III and α-SMA.


Silica promotes the transdifferentiation of rat circulating fibrocytes in vitro.

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

mRNA expression levels of collagen I, collagen III and α-SMA in cFbs treated with the supernatant of AM treated with various concentrations of SiO2. The cFbs were treated with the supernatant of AM treated with SiO2 at concentrations of 0, 20, 40, 60 or 80 µg/ml for 24 h, and the control group was not treated. (A) The cFbs were harvested, total RNA was extracted and the mRNA expression levels were determined by reverse transcription-quantitative polymerase chain reaction. (B) The mRNA expression levels of collagen I, collagen III and α-SMA were quantified and normalized relative to internal β-actin mRNA. The bands were quantified by densitometric analysis. Values are expressed as the mean ± standard deviation from three independent experiments (n=6). *P<0.01, vs. the control group. cFbs, circulating fibrocytes; α-SMA, α smooth muscle actin; SiO2, crystalline silica; AM, alveolar macrophages.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5-mmr-12-04-5828: mRNA expression levels of collagen I, collagen III and α-SMA in cFbs treated with the supernatant of AM treated with various concentrations of SiO2. The cFbs were treated with the supernatant of AM treated with SiO2 at concentrations of 0, 20, 40, 60 or 80 µg/ml for 24 h, and the control group was not treated. (A) The cFbs were harvested, total RNA was extracted and the mRNA expression levels were determined by reverse transcription-quantitative polymerase chain reaction. (B) The mRNA expression levels of collagen I, collagen III and α-SMA were quantified and normalized relative to internal β-actin mRNA. The bands were quantified by densitometric analysis. Values are expressed as the mean ± standard deviation from three independent experiments (n=6). *P<0.01, vs. the control group. cFbs, circulating fibrocytes; α-SMA, α smooth muscle actin; SiO2, crystalline silica; AM, alveolar macrophages.
Mentions: To evaluate the effects of SiO2 on collagen I, collagen III and α-SMA mRNA expression in cFbs, the cFbs were treated with the supernatant of SiO2-treated AM (20, 40, 60 and 80 µg/ml for 24 h). Treatment with the supernatant of AM treated with various concentrations of SiO2 increased the mRNA expression levels of collagen I, collagen III and α-SMA in the cFbs. This increase in expression levels became apparent at 20 µg/ml SiO2 and markedly apparent at 80 µg/ml SiO2. The expression of collagen I, collagen III and α-SMA was significantly and dose-dependently upregulated following treatment with 20, 40 and 60 µg/ml SiO2, as compared with that in the control group (P<0.05), while no statistically significant difference was observed between the 60 µg/ml and 80 µg/ml groups (P>0.05; Fig. 5A and B). Following treatment of the cFbs with the supernatant of AM treated with 80 µg/ml SiO2 for 28 days, the mRNA expression levels of collagen I, collagen III and α-SMA were determined by RT-qPCR. As shown in Fig. 6, increases in collagen I and collagen III expression were detected on day three, and increases in α-SMA expression was observed on day 13 of incubation, whereas in the control and 0 µg/ml groups, elevated collagen I and collagen III expression was detected on day 6, and elevated α-SMA expression on day 19. A significant upregulation of the expression levels of collagen I and collagen III in the treated groups was observed on days 3, 6, 9, 12, 15 and 18 (P<0.05). The expression levels of α-SMA were significantly upregulated on days 13, 16, 19, 22, 25, 28, as compared with those in the control group (P<0.05). These results suggested that SiO2 was able to stimulate collagen I, collagen III and α-SMA activity at the transcriptional level, and increase the mRNA synthesis of collagen I, collagen III and α-SMA.

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