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通过RNA测序和定量PCR证明诺丽对牙龈来源干细胞的细胞活力和成骨分化的影响。

Effects of noni on cellular viability and osteogenic differentiation of gingiva-derived stem cells demonstrated by RNA sequencing and quantitative PCR.

作者信息

Song Young-Min, Lee Hyun-Jin, Min Sae-Kyung, Park Yoon-Hee, Oh Jae-Kwen, Kim Ji-Youn, Park Jun-Beom

机构信息

Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.

Ebiogen, Seoul 04785, Republic of Korea.

出版信息

Exp Ther Med. 2022 Jan;23(1):32. doi: 10.3892/etm.2021.10954. Epub 2021 Nov 8.

DOI:10.3892/etm.2021.10954
PMID:34824640
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8611496/
Abstract

Noni fruit () has been widely used in traditional medicine across tropical and subtropical regions, and is now being paid more attention in Western medicine. The present study aimed to investigate the effects of noni extract on the change in the cellular morphology, maintenance of cellular viability and enhancement of osteogenic differentiation of stem cells. Stem cells obtained from gingiva were cultured where noni extracts existed at concentrations ranging from 10-200 ng/ml. Evaluations of cell morphology and cellular viability were performed. Alkaline phosphatase activity assays were performed to assess the osteogenic differentiation. Alizarin Red S staining was performed to evaluate the calcium deposits in the culture, with the addition of noni extract. Global gene expression was analyzed via next-generation mRNA sequencing. Gene ontology and pathway analyses were performed to determine the associated mechanisms. Validation procedures were performed via quantitative (q)PCR analysis. The addition of noni at concentrations ranging from 10-200 ng/ml did not produce significant morphological changes. There were significantly higher values of cellular viability, with the highest value at 100 ng/ml compared with the control (P<0.05). Furthermore, significantly higher values of alkaline phosphatase activity was noted in the 10 and 100 ng/ml groups compared with the 0 ng/ml group on day 7 (P<0.05). Alizarin Red S staining revealed calcium deposits in each group. In addition, the highest value for Alizarin Red S staining was observed at 100 ng/ml compared with the unloaded control (P<0.05). qPCR analysis demonstrated that the mRNA expression levels of RUNX2, BSP, OCN and COL1A1 increased following treatment with noni. Taken together, the results of the present study suggest that noni extract has enhancing effects on gingiva-derived mesenchymal stem cells, by enhancing cellular viability and osteogenic differentiation.

摘要

诺丽果（）在热带和亚热带地区的传统医学中已被广泛使用，目前在西医中也受到越来越多的关注。本研究旨在探讨诺丽提取物对干细胞细胞形态变化、细胞活力维持以及成骨分化增强的影响。从牙龈中获取的干细胞在存在浓度范围为10 - 200 ng/ml的诺丽提取物的条件下进行培养。对细胞形态和细胞活力进行评估。进行碱性磷酸酶活性测定以评估成骨分化。在添加诺丽提取物的情况下进行茜素红S染色以评估培养物中的钙沉积。通过下一代mRNA测序分析全局基因表达。进行基因本体论和通路分析以确定相关机制。通过定量（q）PCR分析进行验证程序。添加浓度范围为10 - 200 ng/ml的诺丽提取物未产生明显的形态变化。细胞活力值显著更高，与对照组相比，100 ng/ml时的值最高（P<0.05）。此外，在第7天，与0 ng/ml组相比，10和100 ng/ml组的碱性磷酸酶活性值显著更高（P<0.05）。茜素红S染色显示每组均有钙沉积。此外，与未加载对照组相比，在100 ng/ml时观察到茜素红S染色的最高值（P<0.05）。qPCR分析表明，用诺丽处理后RUNX2、BSP、OCN和COL1A1的mRNA表达水平增加。综上所述，本研究结果表明，诺丽提取物通过增强细胞活力和成骨分化对牙龈来源的间充质干细胞具有增强作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4912/8611496/69e67856323b/etm-23-01-10954-g10.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4912/8611496/2d3e9b5b291a/etm-23-01-10954-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4912/8611496/2a8f043499b2/etm-23-01-10954-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4912/8611496/4652a994df0c/etm-23-01-10954-g03.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4912/8611496/60854c7174e4/etm-23-01-10954-g08.jpg
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