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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) Microscopic images of macrophages treated with SiO2 for 24 h and of cFbs treated with macrophage supernatant. (B) Growth curve of cFbs. The survival rate of macrophages stimulated with SiO2 at 0, 20, 40, 60, 80, 100 or 120 µg/ml (n=6 per treatment group) as determined by MTT assay. (C) Assessment the effects of SiO2 on the proliferative rate of the cFbs. The cFbs were seeded and cultured with 80 µg/ml SiO2 treated AM supernatant for 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 days. Viable cells were detected by MTT assay and the cells were counted. Values are expressed as the mean ± standard deviation. *P<0.05, vs. the control group (0 µg/ml SiO2-treated group). cFbs, circulating fibrocytes; AMs, alveolar macrophages; SiO2, crystalline silica.
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f1-mmr-12-04-5828: (A) Microscopic images of macrophages treated with SiO2 for 24 h and of cFbs treated with macrophage supernatant. (B) Growth curve of cFbs. The survival rate of macrophages stimulated with SiO2 at 0, 20, 40, 60, 80, 100 or 120 µg/ml (n=6 per treatment group) as determined by MTT assay. (C) Assessment the effects of SiO2 on the proliferative rate of the cFbs. The cFbs were seeded and cultured with 80 µg/ml SiO2 treated AM supernatant for 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 days. Viable cells were detected by MTT assay and the cells were counted. Values are expressed as the mean ± standard deviation. *P<0.05, vs. the control group (0 µg/ml SiO2-treated group). cFbs, circulating fibrocytes; AMs, alveolar macrophages; SiO2, crystalline silica.

Mentions: The primary culture model of rat AMs and cFbs in vitro is shown in Fig. 1A. Following treatment of macrophages with various concentrations of SiO2 (0, 20, 40, 60, 80, 100 and 120 µg/ml) for 24 h, the cell viability was detected using an MTT assay. As shown in Fig. 1B, compared with the control group (0 µg/ml SiO2), treatment with SiO2 at concentrations of 100 and 120 µg/ml significantly inhibited the growth of AMs, with a significant decrease in cellular activity (P<0.05). To further confirm the effects of SiO2 on the proliferative rate of cFbs, the cFbs were cultured with AM supernatant, treated with 80 µg/ml SiO2 and subjected to an MTT assay at 1 day intervals. The cFbs were seeded and cultured for 1–10 days, and harvested and counted each day. As shown in the growth curve (Fig. 1C), the doubling time of the cells was 96 h, and the proliferative activity of the cFbs reached a peak between the fourth and eighth day.


Silica promotes the transdifferentiation of rat circulating fibrocytes in vitro.

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

(A) Microscopic images of macrophages treated with SiO2 for 24 h and of cFbs treated with macrophage supernatant. (B) Growth curve of cFbs. The survival rate of macrophages stimulated with SiO2 at 0, 20, 40, 60, 80, 100 or 120 µg/ml (n=6 per treatment group) as determined by MTT assay. (C) Assessment the effects of SiO2 on the proliferative rate of the cFbs. The cFbs were seeded and cultured with 80 µg/ml SiO2 treated AM supernatant for 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 days. Viable cells were detected by MTT assay and the cells were counted. Values are expressed as the mean ± standard deviation. *P<0.05, vs. the control group (0 µg/ml SiO2-treated group). cFbs, circulating fibrocytes; AMs, alveolar macrophages; SiO2, crystalline silica.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1-mmr-12-04-5828: (A) Microscopic images of macrophages treated with SiO2 for 24 h and of cFbs treated with macrophage supernatant. (B) Growth curve of cFbs. The survival rate of macrophages stimulated with SiO2 at 0, 20, 40, 60, 80, 100 or 120 µg/ml (n=6 per treatment group) as determined by MTT assay. (C) Assessment the effects of SiO2 on the proliferative rate of the cFbs. The cFbs were seeded and cultured with 80 µg/ml SiO2 treated AM supernatant for 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 days. Viable cells were detected by MTT assay and the cells were counted. Values are expressed as the mean ± standard deviation. *P<0.05, vs. the control group (0 µg/ml SiO2-treated group). cFbs, circulating fibrocytes; AMs, alveolar macrophages; SiO2, crystalline silica.
Mentions: The primary culture model of rat AMs and cFbs in vitro is shown in Fig. 1A. Following treatment of macrophages with various concentrations of SiO2 (0, 20, 40, 60, 80, 100 and 120 µg/ml) for 24 h, the cell viability was detected using an MTT assay. As shown in Fig. 1B, compared with the control group (0 µg/ml SiO2), treatment with SiO2 at concentrations of 100 and 120 µg/ml significantly inhibited the growth of AMs, with a significant decrease in cellular activity (P<0.05). To further confirm the effects of SiO2 on the proliferative rate of cFbs, the cFbs were cultured with AM supernatant, treated with 80 µg/ml SiO2 and subjected to an MTT assay at 1 day intervals. The cFbs were seeded and cultured for 1–10 days, and harvested and counted each day. As shown in the growth curve (Fig. 1C), the doubling time of the cells was 96 h, and the proliferative activity of the cFbs reached a peak between the fourth and eighth day.

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