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羟基红花黄色素A对缺氧和血清剥夺条件下间充质干细胞增殖和凋亡的影响。

Effects of HSYA on the proliferation and apoptosis of MSCs exposed to hypoxic and serum deprivation conditions.

作者信息

Song Xiaoqing, Su Lining, Yin Haifeng, Dai Jin, Wei Huiping

机构信息

Biology Office, Basic Medical College of Hebei North University, Zhangjiakou, Hebei 075000, P.R. China.

出版信息

Exp Ther Med. 2018 Jun;15(6):5251-5260. doi: 10.3892/etm.2018.6125. Epub 2018 May 3.

Abstract

As a primary active ingredient of safflor yellow, hydroxysafflor yellow A (HSYA) exhibits notable antioxidative and neuroprotective effects. The aim of the present study was to investigate the protective effects of HSYA in mesenchymal stem cells (MSCs) exposed to hypoxia (5% O) and serum deprivation (H/SD), and to explore the mechanisms underlying HSYA-mediated protection. Under H/SD conditions, HSYA was applied to protect MSCs against injury. Cell viability, proliferation, apoptosis and reactive oxygen species (ROS) levels were determined using an 5-ethynyl-2'-deoxyuridine assay, MTT assay, Hoechst 33342/propidium iodide and 2',7'-dichlorodihydrofluorescein diacetate staining, respectively. The results revealed that 160 mg/l HSYA significantly reduced apoptosis and ROS levels compared with the H/SD group; however, HSYA demonstrated minimal effects on cell proliferation. A western blot assay demonstrated that HSYA reduced cleaved caspase-3 expression and cytC release from the mitochondria to the cytoplasm when compared with the H/SD group. In addition, western blotting and RT-qPCR analyses revealed that HSYA treatment significantly increased the expression of hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF). In conclusion, the results of the current study demonstrated that HSYA exerts protective effects against H/SD-induced apoptosis in MSCs potentially via activation of the HIF-1α/VEGF signaling pathway and stabilization of the mitochondrial membrane.

摘要

作为红花黄色素的主要活性成分,羟基红花黄色素A(HSYA)具有显著的抗氧化和神经保护作用。本研究旨在探讨HSYA对暴露于低氧(5%氧气)和血清剥夺(H/SD)环境中的间充质干细胞(MSCs)的保护作用,并探究HSYA介导保护作用的潜在机制。在H/SD条件下,应用HSYA保护MSCs免受损伤。分别使用5-乙炔基-2'-脱氧尿苷检测法、MTT检测法、Hoechst 33342/碘化丙啶以及2',7'-二氯二氢荧光素二乙酸酯染色法测定细胞活力、增殖、凋亡和活性氧(ROS)水平。结果显示,与H/SD组相比,160mg/l的HSYA显著降低了细胞凋亡和ROS水平;然而,HSYA对细胞增殖的影响极小。蛋白质免疫印迹分析表明,与H/SD组相比,HSYA降低了裂解的半胱天冬酶-3的表达以及细胞色素C从线粒体向细胞质的释放。此外,蛋白质免疫印迹和RT-qPCR分析显示,HSYA处理显著增加了缺氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)的表达。总之,本研究结果表明,HSYA可能通过激活HIF-1α/VEGF信号通路和稳定线粒体膜,对H/SD诱导的MSCs凋亡发挥保护作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/00f6/5996714/c512ce9f2d5a/etm-15-06-5251-g00.jpg

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