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The Anti-fibrotic Effects and Mechanisms of MicroRNA-486-5p in Pulmonary Fibrosis.

Ji X, Wu B, Fan J, Han R, Luo C, Wang T, Yang J, Han L, Zhu B, Wei D, Chen J, Ni C - Sci Rep (2015)

Bottom Line: Notably, miR-486-5p levels were decreased in the serum samples of patients with silicosis, as well as in the lung tissues of patients with silicosis and idiopathic pulmonary fibrosis (IPF).At day 28, miR-486-5p expression significantly decreased both the distribution and severity of lung lesions compared with the silica group (P < 0.01).In addition, miR-486-5p had a similar effect in the BLM group (P < 0.001).

View Article: PubMed Central - PubMed

Affiliation: Department of Occupational Medicine and Environmental Health, School of Public Health, Nanjing Medical University, Nanjing, China.

ABSTRACT
To identify microRNAs (miRNAs, miRs) with potential roles in lung fibrogenesis, we performed genome-wide profiling of miRNA expression in lung tissues from a silica-induced mouse model of pulmonary fibrosis using microarrays. Seventeen miRNAs were selected for validation via qRT-PCR based on the fold changes between the silica and the control group. The dysregulation of five miRNAs, including miR-21, miR-455, miR-151-3p, miR-486-5p and miR-3107, were confirmed by qRT-PCRs in silica-induced mouse model of pulmonary fibrosis and were also confirmed in a bleomycin (BLM)-induced mouse lung fibrosis. Notably, miR-486-5p levels were decreased in the serum samples of patients with silicosis, as well as in the lung tissues of patients with silicosis and idiopathic pulmonary fibrosis (IPF). In addition, as determined by luciferase assays and Western blotting, SMAD2, a crucial mediator of pulmonary fibrosis, was identified to be one of target genes of miR-486-5p. To test the potential therapeutic significance of this miRNA, we overexpressed miR-486-5p in animal models. At day 28, miR-486-5p expression significantly decreased both the distribution and severity of lung lesions compared with the silica group (P < 0.01). In addition, miR-486-5p had a similar effect in the BLM group (P < 0.001). These results indicate that miR-486-5p may inhibit fibrosis.

No MeSH data available.


Related in: MedlinePlus

MiR-486-5p is down-regulated in the lung tissues of patients with silicosis and IPF, as well as serum from patients with silicosis.(A) A heat map representing the differentially expressed microRNAs in the lungs of the C57BL/6 mice in response to silica at the indicated time points. The up-regulated microRNAs are indicated in progressively brighter shades of red, and the down-regulated microRNAs are indicated in progressively brighter shades of green. (B) Real-time PCR validation of the microarray results from the lungs of mice with silica-induced pulmonary fibrosis. The mice were injected intratracheally with either saline or silica (50 mg/kg of silica in 0.05 ml of sterile saline) and sacrificed on days 0, 3, 7, 14, 28 and 56 after injection (n = 6 mice in each group). The data are expressed as the mean ± SD. *P < 0.05, #P < 0.01. (C) The mice were injected intratracheally with either saline or bleomycin (1.5 U/kg of bleomycin in 0.05 ml of sterile saline) and sacrificed on days 0, 3, 7, 14 and 28 after injection. The five validated miRNAs were consistent with the microarray results from the BLM-induced pulmonary fibrosis model as determined via a real-time PCR analysis. U6 was used as an internal control (n = 6 mice in each group). The data are expressed as the mean ± SD. *P < 0.05, #P < 0.01. (D) The serum concentrations of the five validated miRNAs were measured in 60 patients with silicosis (n = 20 per stage I, II, III) and 20 normal control subjects by qRT-PCR. The data are expressed as the mean ± SD. *P < 0.05. (E) miR-486-5p expression in the lungs of patients with either silicosis (n = 5) or IPF (n = 5) was measured by qRT-PCR. The data are expressed as the mean ± SD. *P < 0.05 compared with the normal control lungs (n = 2).
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f1: MiR-486-5p is down-regulated in the lung tissues of patients with silicosis and IPF, as well as serum from patients with silicosis.(A) A heat map representing the differentially expressed microRNAs in the lungs of the C57BL/6 mice in response to silica at the indicated time points. The up-regulated microRNAs are indicated in progressively brighter shades of red, and the down-regulated microRNAs are indicated in progressively brighter shades of green. (B) Real-time PCR validation of the microarray results from the lungs of mice with silica-induced pulmonary fibrosis. The mice were injected intratracheally with either saline or silica (50 mg/kg of silica in 0.05 ml of sterile saline) and sacrificed on days 0, 3, 7, 14, 28 and 56 after injection (n = 6 mice in each group). The data are expressed as the mean ± SD. *P < 0.05, #P < 0.01. (C) The mice were injected intratracheally with either saline or bleomycin (1.5 U/kg of bleomycin in 0.05 ml of sterile saline) and sacrificed on days 0, 3, 7, 14 and 28 after injection. The five validated miRNAs were consistent with the microarray results from the BLM-induced pulmonary fibrosis model as determined via a real-time PCR analysis. U6 was used as an internal control (n = 6 mice in each group). The data are expressed as the mean ± SD. *P < 0.05, #P < 0.01. (D) The serum concentrations of the five validated miRNAs were measured in 60 patients with silicosis (n = 20 per stage I, II, III) and 20 normal control subjects by qRT-PCR. The data are expressed as the mean ± SD. *P < 0.05. (E) miR-486-5p expression in the lungs of patients with either silicosis (n = 5) or IPF (n = 5) was measured by qRT-PCR. The data are expressed as the mean ± SD. *P < 0.05 compared with the normal control lungs (n = 2).

Mentions: To validate the results of our miRNA profiling experiment, we confirmed the differential expression of these miRNAs via real-time quantitative reverse transcription polymerase chain reactions (qRT-PCRs). Seventeen miRNAs with raw expression values greater than 100 were selected from among the 62 miRNA molecules with altered expression levels (Fig. 1A). Five miRNAs, miR-21, miR-455, miR-151-3p, miR-486-5p and miR-3107, were validated to be dysregulated in the lung tissues from mice with silica-induced fibrosis (n = 6 per group) (Fig. 1B). Because fibrotic diseases may share common pathogenic mechanisms181920, we sought to validate our findings in another experimental model of pulmonary fibrosis (induced by BLM), as well as in human fibrotic diseases. We found that miR-21, miR-455, miR-151-3p, miR-486-5p and miR-3107 were differentially expressed in a BLM-induced mouse model of pulmonary fibrosis (Fig. 1C). In serum samples obtained from patients with silicosis, miR-486-5p expression was significantly decreased in all cohorts (stages I, II and III, P < 0.05) compared with the control subjects (Fig. 1D). The lung tissues obtained from the patients with either silicosis or IPF also exhibited decreased miR-486-5p expression (Fig. 1E).


The Anti-fibrotic Effects and Mechanisms of MicroRNA-486-5p in Pulmonary Fibrosis.

Ji X, Wu B, Fan J, Han R, Luo C, Wang T, Yang J, Han L, Zhu B, Wei D, Chen J, Ni C - Sci Rep (2015)

MiR-486-5p is down-regulated in the lung tissues of patients with silicosis and IPF, as well as serum from patients with silicosis.(A) A heat map representing the differentially expressed microRNAs in the lungs of the C57BL/6 mice in response to silica at the indicated time points. The up-regulated microRNAs are indicated in progressively brighter shades of red, and the down-regulated microRNAs are indicated in progressively brighter shades of green. (B) Real-time PCR validation of the microarray results from the lungs of mice with silica-induced pulmonary fibrosis. The mice were injected intratracheally with either saline or silica (50 mg/kg of silica in 0.05 ml of sterile saline) and sacrificed on days 0, 3, 7, 14, 28 and 56 after injection (n = 6 mice in each group). The data are expressed as the mean ± SD. *P < 0.05, #P < 0.01. (C) The mice were injected intratracheally with either saline or bleomycin (1.5 U/kg of bleomycin in 0.05 ml of sterile saline) and sacrificed on days 0, 3, 7, 14 and 28 after injection. The five validated miRNAs were consistent with the microarray results from the BLM-induced pulmonary fibrosis model as determined via a real-time PCR analysis. U6 was used as an internal control (n = 6 mice in each group). The data are expressed as the mean ± SD. *P < 0.05, #P < 0.01. (D) The serum concentrations of the five validated miRNAs were measured in 60 patients with silicosis (n = 20 per stage I, II, III) and 20 normal control subjects by qRT-PCR. The data are expressed as the mean ± SD. *P < 0.05. (E) miR-486-5p expression in the lungs of patients with either silicosis (n = 5) or IPF (n = 5) was measured by qRT-PCR. The data are expressed as the mean ± SD. *P < 0.05 compared with the normal control lungs (n = 2).
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4569899&req=5

f1: MiR-486-5p is down-regulated in the lung tissues of patients with silicosis and IPF, as well as serum from patients with silicosis.(A) A heat map representing the differentially expressed microRNAs in the lungs of the C57BL/6 mice in response to silica at the indicated time points. The up-regulated microRNAs are indicated in progressively brighter shades of red, and the down-regulated microRNAs are indicated in progressively brighter shades of green. (B) Real-time PCR validation of the microarray results from the lungs of mice with silica-induced pulmonary fibrosis. The mice were injected intratracheally with either saline or silica (50 mg/kg of silica in 0.05 ml of sterile saline) and sacrificed on days 0, 3, 7, 14, 28 and 56 after injection (n = 6 mice in each group). The data are expressed as the mean ± SD. *P < 0.05, #P < 0.01. (C) The mice were injected intratracheally with either saline or bleomycin (1.5 U/kg of bleomycin in 0.05 ml of sterile saline) and sacrificed on days 0, 3, 7, 14 and 28 after injection. The five validated miRNAs were consistent with the microarray results from the BLM-induced pulmonary fibrosis model as determined via a real-time PCR analysis. U6 was used as an internal control (n = 6 mice in each group). The data are expressed as the mean ± SD. *P < 0.05, #P < 0.01. (D) The serum concentrations of the five validated miRNAs were measured in 60 patients with silicosis (n = 20 per stage I, II, III) and 20 normal control subjects by qRT-PCR. The data are expressed as the mean ± SD. *P < 0.05. (E) miR-486-5p expression in the lungs of patients with either silicosis (n = 5) or IPF (n = 5) was measured by qRT-PCR. The data are expressed as the mean ± SD. *P < 0.05 compared with the normal control lungs (n = 2).
Mentions: To validate the results of our miRNA profiling experiment, we confirmed the differential expression of these miRNAs via real-time quantitative reverse transcription polymerase chain reactions (qRT-PCRs). Seventeen miRNAs with raw expression values greater than 100 were selected from among the 62 miRNA molecules with altered expression levels (Fig. 1A). Five miRNAs, miR-21, miR-455, miR-151-3p, miR-486-5p and miR-3107, were validated to be dysregulated in the lung tissues from mice with silica-induced fibrosis (n = 6 per group) (Fig. 1B). Because fibrotic diseases may share common pathogenic mechanisms181920, we sought to validate our findings in another experimental model of pulmonary fibrosis (induced by BLM), as well as in human fibrotic diseases. We found that miR-21, miR-455, miR-151-3p, miR-486-5p and miR-3107 were differentially expressed in a BLM-induced mouse model of pulmonary fibrosis (Fig. 1C). In serum samples obtained from patients with silicosis, miR-486-5p expression was significantly decreased in all cohorts (stages I, II and III, P < 0.05) compared with the control subjects (Fig. 1D). The lung tissues obtained from the patients with either silicosis or IPF also exhibited decreased miR-486-5p expression (Fig. 1E).

Bottom Line: Notably, miR-486-5p levels were decreased in the serum samples of patients with silicosis, as well as in the lung tissues of patients with silicosis and idiopathic pulmonary fibrosis (IPF).At day 28, miR-486-5p expression significantly decreased both the distribution and severity of lung lesions compared with the silica group (P < 0.01).In addition, miR-486-5p had a similar effect in the BLM group (P < 0.001).

View Article: PubMed Central - PubMed

Affiliation: Department of Occupational Medicine and Environmental Health, School of Public Health, Nanjing Medical University, Nanjing, China.

ABSTRACT
To identify microRNAs (miRNAs, miRs) with potential roles in lung fibrogenesis, we performed genome-wide profiling of miRNA expression in lung tissues from a silica-induced mouse model of pulmonary fibrosis using microarrays. Seventeen miRNAs were selected for validation via qRT-PCR based on the fold changes between the silica and the control group. The dysregulation of five miRNAs, including miR-21, miR-455, miR-151-3p, miR-486-5p and miR-3107, were confirmed by qRT-PCRs in silica-induced mouse model of pulmonary fibrosis and were also confirmed in a bleomycin (BLM)-induced mouse lung fibrosis. Notably, miR-486-5p levels were decreased in the serum samples of patients with silicosis, as well as in the lung tissues of patients with silicosis and idiopathic pulmonary fibrosis (IPF). In addition, as determined by luciferase assays and Western blotting, SMAD2, a crucial mediator of pulmonary fibrosis, was identified to be one of target genes of miR-486-5p. To test the potential therapeutic significance of this miRNA, we overexpressed miR-486-5p in animal models. At day 28, miR-486-5p expression significantly decreased both the distribution and severity of lung lesions compared with the silica group (P < 0.01). In addition, miR-486-5p had a similar effect in the BLM group (P < 0.001). These results indicate that miR-486-5p may inhibit fibrosis.

No MeSH data available.


Related in: MedlinePlus