Differential Expression Profiling of Salivary Exosomal microRNAs in a Single Case of Periodontitis - A Pilot Study

서지정보

ㆍ발행기관 : 대한구강생물학회 ㆍ수록지정보 : International Journal of Oral Biology / 43권 / 4호
ㆍ저자명 : Sung Nam Park, Young Woo Son, Eun Joo Choi, Hyung-Keun You, Min Seuk Kim

목차

Introduction
Materials and Methods
Clinical subject and ethics statement
Saliva collection and isolation of salivary exosomes
Reagents
Cell culture and transfection of miRNA mimics
Affymetrix miRNA arrays
MicroRNA target prediction and pathway analysis
Reverse-transcription polymerase chain-reaction(PCR)
Western blot
Statistical analysis
Results
Clinical characteristics
Characteristics of differentially expressed miRNAsin salivary exosomes
Overexpression of miRNA mimics (hsa-miR-4487,-4532, and 7108-5p) in human gingival fibroblastsresults in induction of IL-6 expression and p38activation
Discussion
References

영어 초록

Exosomes are Nano-sized lipid vesicles secreted from mammalian cells containing diverse cellular materials such as proteins, lipids, and nucleotides. Multiple lines of evidence indicate that in saliva, exosomes and their contents such as microRNAs (miRNAs) mediate numerous cellular responses upon delivery to recipient cells. The objective of this study was to characterize the different expression profile of exosomal miRNAs in saliva samples, periodically isolated from a single periodontitis patient. Unstimulated saliva was collected from a single patient over time periods for managing periodontitis. MicroRNAs extracted from each phase were investigated for the expression of exosomal miRNAs. Salivary exosomal miRNAs were analyzed using Affymetrix miRNA arrays and prediction of target genes and pathways for its different expression performed using DIANA-mirPath, a web-based, computational tool. Following the delivery of miRNA mimics (hsa-miR-4487, -4532, and -7108-5p) into human gingival fibroblasts, the expression of pro-inflammatory cytokines and activation of the MAPK pathway were evaluated through RT-PCR and western blotting. In each phase, 13 and 43 miRNAs were found to be differently expressed (|FC| ≥ 2). Among these, hsa-miR-4487 (|FC|=9.292005) and hasmiR- 4532 (|FC|=18.322697) were highly up-regulated in the clinically severe phase, whereas hsa-miR-7108-5p (|FC|= 12.20601) was strongly up-regulated in the clinically mild phase. In addition, the overexpression of miRNA mimics in human gingival fibroblasts resulted in a significant induction of IL-6 mRNA expression and p38 phosphorylation. The findings of this study established alterations in salivary exosomal miRNAs which are dependent on the severity of periodontitis and may act as potential candidates for the treatment of oral inflammatory diseases.

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