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绿色光生物调节对人脂肪间充质干细胞的抗炎作用。

Anti-inflammatory effect of green photobiomodulation in human adipose-derived mesenchymal stem cells.

机构信息

Tissue Engineering and Regenerative Medicine Institute, Stem Cell Research Center, Central Tehran Branch, Islamic Azad University, Tehran, Iran.

Department of Biology, Faculty of Science, Central Tehran Branch, Islamic Azad University, Tehran, Iran.

出版信息

Lasers Med Sci. 2022 Dec;37(9):3693-3703. doi: 10.1007/s10103-022-03654-5. Epub 2022 Nov 17.

Abstract

Photo biomodulation (PBM) as a non-invasive and safe treatment has been demonstrated the anti-inflammatory potential in a variety of cell types, including stem cells. However, further investigations using different laser parameters combined with more accurate methods such as quantitative measurement of inflammatory gene expression at the mRNA level are still necessary. The aim of this study was to evaluate the effect of 532 nm green laser on cell proliferation as well as expression of inflammatory genes in human adipose-derived mesenchymal stem cells (hADMSCs) using RNA sequencing (RNA-seq) technique and confirmatory RT-PCR. hADMSCs were cultured in DMEM low glocuse medium with 10% fetal bovine serum until the fourth passage. Cultured cells were divided in two groups: control group (no laser irradiation) and laser group, irradiated with 532 nm laser at 44 m J/cm with an output power of 50 mW and a density of 6 mW/cm, every other day, 7 s each time. The cell viability was assessed using MTT assay 24 h after each irradiation on days 3, 5, and 7 after cell seeding, followed by performing RNA-seq and RT-PCR. The MTT assay showed that PBM increased cell proliferation on day 5 after irradiation compared to day 3 and decreased on day 7 compared to day 5. In addition, gene expression analysis in hADMSCs using RNA-seq revealed down-regulation of inflammatory genes including CSF2, CXCL2, 3, 5, 6, 8, and CCL2, 7. These results indicate that 532 nm PBM with the parameters used in this study has a time-dependent effect on hADMSCs proliferation as well as anti-inflammatory potential.

摘要

光生物调节(PBM)作为一种非侵入性和安全的治疗方法,已在多种细胞类型中显示出抗炎潜力,包括干细胞。然而,仍需要使用不同的激光参数并结合更准确的方法(例如在 mRNA 水平上定量测量炎症基因的表达)进行进一步研究。本研究旨在使用 RNA 测序(RNA-seq)技术和确认性 RT-PCR 评估 532nm 绿光对人脂肪间充质干细胞(hADMSCs)细胞增殖和炎症基因表达的影响。hADMSCs 在含 10%胎牛血清的 DMEM 低糖培养基中培养,直至第 4 代。培养的细胞分为两组:对照组(无激光照射)和激光组,用 532nm 激光照射,输出功率为 50mW,密度为 6mW/cm,每隔一天照射一次,每次照射 7s。在细胞接种后第 3、5 和 7 天,每次照射后 24h 通过 MTT 测定法评估细胞活力,然后进行 RNA-seq 和 RT-PCR。MTT 测定法显示,与第 3 天相比,PBM 在照射后第 5 天增加了细胞增殖,而与第 5 天相比,在第 7 天减少了细胞增殖。此外,使用 RNA-seq 对 hADMSCs 的基因表达分析显示,炎症基因 CSF2、CXCL2、3、5、6、8 和 CCL2、7 的表达下调。这些结果表明,在本研究中使用的参数下,532nm PBM 对 hADMSCs 的增殖和抗炎潜力具有时间依赖性的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd05/9668707/916d9eac0d12/10103_2022_3654_Fig1_HTML.jpg

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