2018PubMedRequires access

Study on the role of Hsa-miR-382-5p in epidural fibrosis.

Lin Ca, Duan Ky, X-W Wang, Zhang Zs

Open publisher page 9 citations

Abstract

OBJECTIVE: To investigate the role and potential mechanism of human serum albumin (hsa)-micro ribonucleic acid (miR)-382-5p in epidural fibrosis formation after laminectomy. MATERIALS AND METHODS: Cells were transfected with miR-382-5p mimic or miR-382-5p inhibitor. Then, 3-(4,5)-dimethylthiahiazol (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay was employed to detect the effect of miR-382-5p on proliferation, and Real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to examine the expressions of miR-382-5p and fibrosis-related proteins after treatment with transforming growth factor beta (TGF-β). Luciferase assay and immunofluorescent staining were done to confirm whether collagen I A1 is a target of miR-382-5p. RESULTS: MTT assay demonstrated that miR-382-5p had no significant effect on fibroblast proliferation. Expressions of miR-382-5p and fibrosis-related proteins were remarkably increased after TGF-β treatment. Collagen I A1 was acknowledged as a target of miR-382-5p. MiR-382-5p mimic statistically enhanced the level of collagen I A1, and miR-382-5p enhanced the expressions of collagen I A1. CONCLUSIONS: Increased miR-382-5p promotes epidural fibrosis by increasing collagen I A1 expression, and miR-382-5p may be a potential novel molecular target for the treatment of epidural fibrosis.

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What this paper is about

OBJECTIVE: To investigate the role and potential mechanism of human serum albumin (hsa)-micro ribonucleic acid (miR)-382-5p in epidural fibrosis formation after laminectomy. MATERIALS AND METHODS: Cells were transfected with miR-382-5p mimic or miR-382-5p inhibitor. Then, 3-(4,5)-dimethylthiahiazol (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay was employed to detect the effect of miR-382-5p on proliferation, and Real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to examine the expressions of miR-382-5p and fibrosis-related proteins after treatment with transforming growth factor beta (TGF-β). Luciferase assay and immunofluorescent staining were done to confirm whether collagen I A1 is a target of miR-382-5p. RESULTS: MTT assay demonstrated that miR-382-5p had no significant effect on fibroblast proliferation. Expressions of miR-382-5p and fibrosis-related proteins were remarkably increased after TGF-β treatment. Collagen I A1 was acknowledged as a target of miR-382-5p. MiR-382-5p mimic statistically enhanced the level of collagen I A1, and miR-382-5p enhanced the expressions of collagen I A1. CONCLUSIONS: Increased miR-382-5p promotes epidural fibrosis by increasing collagen I A1 expression, and miR-382-5p may be a potential novel molecular target for the treatment of epidural fibrosis.

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Available abstract

OBJECTIVE: To investigate the role and potential mechanism of human serum albumin (hsa)-micro ribonucleic acid (miR)-382-5p in epidural fibrosis formation after laminectomy. MATERIALS AND METHODS: Cells were transfected with miR-382-5p mimic or miR-382-5p inhibitor. Then, 3-(4,5)-dimethylthiahiazol (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay was employed to detect the effect of miR-382-5p on proliferation, and Real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to examine the expressions of miR-382-5p and fibrosis-related proteins after treatment with transforming growth factor beta (TGF-β). Luciferase assay and immunofluorescent staining were done to confirm whether collagen I A1 is a target of miR-382-5p. RESULTS: MTT assay demonstrated that miR-382-5p had no significant effect on fibroblast proliferation. Expressions of miR-382-5p and fibrosis-related proteins were remarkably increased after TGF-β treatment. Collagen I A1 was acknowledged as a target of miR-382-5p. MiR-382-5p mimic statistically enhanced the level of collagen I A1, and miR-382-5p enhanced the expressions of collagen I A1. CONCLUSIONS: Increased miR-382-5p promotes epidural fibrosis by increasing collagen I A1 expression, and miR-382-5p may be a potential novel molecular target for the treatment of epidural fibrosis.

Key concepts: Fibrosis, Chemistry, Blot, MTT assay, Fibroblast, Transfection, Transforming growth factor, Luciferase

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