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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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Related in: MedlinePlus

cFbs were identified by flow cytometry after 6 weeks of culture. Representative flow cytometric dot plots of circulating fibrocytes. (A) Isotype control for collagen 1 and CD45 using unstained cells. (B) Positive population for collagen 1 and CD45. COLI, collagen I; PE, phycoerythrin; FITC, fluorescein isothiocyanate; cFbs, circulating fibrocytes.
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f2-mmr-12-04-5828: cFbs were identified by flow cytometry after 6 weeks of culture. Representative flow cytometric dot plots of circulating fibrocytes. (A) Isotype control for collagen 1 and CD45 using unstained cells. (B) Positive population for collagen 1 and CD45. COLI, collagen I; PE, phycoerythrin; FITC, fluorescein isothiocyanate; cFbs, circulating fibrocytes.

Mentions: cFbs were identified by flow cytometry, and the purity of the cFbs was assayed by staining with collagen I and CD45 antibodies after six weeks of culture. The results demonstrated that 99% of the cultured cells co-expressed collagen I and CD45 (Fig. 2).


Silica promotes the transdifferentiation of rat circulating fibrocytes in vitro.

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

cFbs were identified by flow cytometry after 6 weeks of culture. Representative flow cytometric dot plots of circulating fibrocytes. (A) Isotype control for collagen 1 and CD45 using unstained cells. (B) Positive population for collagen 1 and CD45. COLI, collagen I; PE, phycoerythrin; FITC, fluorescein isothiocyanate; cFbs, circulating fibrocytes.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2-mmr-12-04-5828: cFbs were identified by flow cytometry after 6 weeks of culture. Representative flow cytometric dot plots of circulating fibrocytes. (A) Isotype control for collagen 1 and CD45 using unstained cells. (B) Positive population for collagen 1 and CD45. COLI, collagen I; PE, phycoerythrin; FITC, fluorescein isothiocyanate; cFbs, circulating fibrocytes.
Mentions: cFbs were identified by flow cytometry, and the purity of the cFbs was assayed by staining with collagen I and CD45 antibodies after six weeks of culture. The results demonstrated that 99% of the cultured cells co-expressed collagen I and CD45 (Fig. 2).

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