Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Pulmonary activation of vitamin D 3 and preventive effect against interstitial pneumonia

doi: 10.3164/jcbn.19-48

Figure Lengend Snippet: Expression of the vitamin D receptor and vitamin D-metabolizing enzymes in human fibroblasts, and VD 3 -dependent suppression of the induction of IL-1β gene expression in pulmonary fibroblasts by bleomycin. (A) To assess gene expression in MRC-5 and MRC-5 SV1 TG1 cells, RT-PCR was performed using specific primers for the vitamin D receptor (VDR) or vitamin D-metabolizing enzymes, including CYP27A1 (27A1), CYP2R1 (2R1), and CYP27B1 (27B1). (B) MRC-5 SV1 TG1 cells were treated with 25 µg/ml bleomycin for 48 h and RT-PCR was performed using specific primers for αSMA, IL-1β, and GAPDH. (C, D) Band intensities of the PCR products were converted to numerical data by image analysis software, and the data for αSMA and IL-1β were normalized by the GAPDH value.

Article Snippet: A normal human fetal pulmonary fibroblast cell line (MRC-5) and an immortalized cell line derived from MRC-5 (MRC-5 SV1 TG1) (KAC Co., Ltd., Kyoto, Japan) were maintained in α-minimal essential medium (α-MEM) containing 10% fetal calf serum (FCS) and antibiotics (Life Technologies Japan Ltd., Tokyo, Japan) at 37°C under 5% CO 2 .

Techniques: Expressing, Gene Expression, Reverse Transcription Polymerase Chain Reaction, Software

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Pulmonary activation of vitamin D 3 and preventive effect against interstitial pneumonia

doi: 10.3164/jcbn.19-48

Figure Lengend Snippet: Pretreatment of pulmonary fibroblasts with VD 3 suppresses bleomycin-induced expression of pro-fibrotic genes, fibrosis markers, and inflammatory markers. (A) MRC-5 SV1 TG1 cells underwent pretreatment with VD 3 or 1,25(OH) 2 D 3 overnight and then were incubated with 25 µg/ml bleomycin for 24 h, after which RT-PCR was performed using specific primers for αSMA, COLA2, SPP1, IL-1β, TGF-β1, and β-actin (ACTB). (B–D) Band intensities of the PCR products were converted to numerical data by image analysis software, and the data for SPP1, IL-1β, and TGF-β1 were normalized by the β-actin value.

Article Snippet: A normal human fetal pulmonary fibroblast cell line (MRC-5) and an immortalized cell line derived from MRC-5 (MRC-5 SV1 TG1) (KAC Co., Ltd., Kyoto, Japan) were maintained in α-minimal essential medium (α-MEM) containing 10% fetal calf serum (FCS) and antibiotics (Life Technologies Japan Ltd., Tokyo, Japan) at 37°C under 5% CO 2 .

Techniques: Expressing, Incubation, Reverse Transcription Polymerase Chain Reaction, Software