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双酚A和光生物调节疗法对脂肪组织来源干细胞中自噬相关基因诱导的影响。

The Effect of bisphenol A and Photobiomodulation Therapy on Autophagy-Related Genes Induction in Adipose Tissue-Derived Stem Cells.

作者信息

Ebrahimi-Kia Yasaman, Noori-Zadeh Ali, Rajaei Farzad, Darabi Shahram, Darabi Leila, Ghasemi Hamidabadi Hatef

机构信息

Student Research Committee, Qazvin University of Medical Sciences, Qazvin, Iran.

Department of Clinical Biochemistry, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.

出版信息

J Lasers Med Sci. 2022 Apr 4;13:e15. doi: 10.34172/jlms.2022.15. eCollection 2022.

Abstract

As adipose tissue-derived stem cells (ADSCs) can divide rapidly and be prepared non-invasively, they have extensively been used in regenerative medicine. On the other hand, a new method of therapy, known as photobiomodulation (PHT), has been used to treat many diseases, such as inflammatory conditions, wound healing and pain. Besides, exposure to chemical substances such as bisphenol A (BPA), at low levels, can lead to autophagy. This study investigated the effects of BPA and PHT on the expression of autophagy-related genes, including LC3, NRF2, P62, in rat ADSCs as a model. ADSCs isolation and purification were confirmed by immunocytochemistry (ICC). The cells were then treated with different concentrations of BPA and also subjected to PHT. Reverse transcription polymerase chain reaction (RT-PCR) was used for the evaluation of LC3, NRF2 and P62 gene expressions. Oil red O staining was used for adipogenic vacuole formation. ICC showed that the isolated cells were CD 49-positive but CD 31 and CD 34-negative. The viability test indicated that the number of live cells after 24 hours in the BPA groups at concentrations of 0, 1, 50, 100 and 200 μM was 100%, 93%, 81%, 72%, and 43% respectively. The difference in cell viability between groups 50, 100 and 200 μM was significant as compared with the control groups ( < 0.05). Moreover, in the group with 1 μM concentration of BPA, the expressions of LC3, NRF2 and P62 genes were upregulated. However, in the treatment group at the concentration of 200 μM of BPA, the LC3 gene was expressed, but NRF2 and P62 genes were downregulated. BPA and PHT induce autophagy and adiposeness in ADSCs in a dose-dependent manner.

摘要

由于脂肪组织来源的干细胞(ADSCs)能够快速分裂且可通过非侵入性方式获取,它们已在再生医学中得到广泛应用。另一方面,一种名为光生物调节(PHT)的新治疗方法已被用于治疗多种疾病,如炎症、伤口愈合和疼痛。此外,低水平接触双酚A(BPA)等化学物质会导致自噬。本研究以大鼠ADSCs为模型,研究了BPA和PHT对自噬相关基因(包括LC3、NRF2、P62)表达的影响。通过免疫细胞化学(ICC)确认了ADSCs的分离和纯化。然后用不同浓度的BPA处理细胞,并对其进行PHT处理。采用逆转录聚合酶链反应(RT-PCR)评估LC3、NRF2和P62基因的表达。用油红O染色观察脂肪生成液泡的形成。ICC显示分离出的细胞CD 49阳性,但CD 31和CD 34阴性。活力测试表明,在0、1、50、100和200 μM浓度的BPA组中,24小时后活细胞数量分别为100%、93%、81%、72%和43%。与对照组相比,50、100和200 μM组之间的细胞活力差异显著(<0.05)。此外,在1 μM浓度的BPA组中,LC3、NRF2和P62基因的表达上调。然而,在200 μM浓度的BPA治疗组中,LC3基因表达,但NRF2和P62基因下调。BPA和PHT以剂量依赖的方式诱导ADSCs中的自噬和脂肪生成。

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