白藜芦醇通过抑制铁死亡保护骨髓间充质干细胞

口腔生物医学 ›› 2023, Vol. ›› Issue (3): 162-168.

白藜芦醇通过抑制铁死亡保护骨髓间充质干细胞

兰冬梅¹,姚超¹,李雪¹,刘海霞²,祁胜财³

1. 安徽理工大学医学院，淮南 232001
2. 同济大学医学院附属上海市第十人民医院口腔科，上海 200072
3. 复旦大学附属口腔医院（筹），上海市口腔医院上海 200000

收稿日期:2022-07-28 修回日期:2022-08-30 出版日期:2023-09-25 发布日期:2023-09-21
通讯作者: 祁胜财 E-mail:dentistqi@163.com
基金资助:
上海市科学技术委员会;上海市科学技术委员会

Resveratrol protects bone marrow mesenchymal stem cells by inhibiting ferroptosis

Received:2022-07-28 Revised:2022-08-30 Online:2023-09-25 Published:2023-09-21

摘要/Abstract

摘要： 目的：探索白藜芦醇（Res）在体外对氧化应激状态下大鼠骨髓间充质干细胞（BMSCs）的保护作用。方法：使用200 μmol/L双氧水处理大鼠BMSCs，使其进入氧化应激状态，采用CCK8、活性氧（ROS）荧光探针、实时荧光定量PCR、Western blot和碱性磷酸酶（ALP）染色等方法分别检测不同浓度的Res对大鼠BMSCs细胞活力、细胞内活性氧水平、成骨分化和铁死亡的影响。结果：Res在1～25 μmol/L浓度范围内对氧化应激状态下的大鼠BMSCs细胞活力有促进作用（P＜0.05）。10 μmol/L Res显著降低了氧化应激状态下的大鼠BMSCs细胞内ROS水平，增加了核红细胞相关因子2（NRF2）、过氧化氢酶（CAT）、超氧化物歧化酶1（SOD-1）和超氧化物歧化酶2（SOD-2）等抗氧化相关基因的表达（P＜0.05）。10 μmol/L Res促进了氧化应激状态下的大鼠BMSCs成骨相关标志物的表达（P＜0.05），明显增强了其ALP染色，促进了铁死亡相关标志物谷胱甘肽过氧化物酶（GPX4）和胱氨酸转运蛋白（XCT）的表达（P＜0.05），降低了脂酰辅酶A合成酶长链家族成员4（ACSL4）、前列腺素内过氧化物合成酶2（PTGS2）和铁蛋白自噬核受体共激活因子4（NCOA4）的表达（P＜0.05）。结论：Res能够降低氧化应激诱导的大鼠BMSCs细胞内活性氧和铁死亡水平，并促进氧化应激状态下大鼠BMSCs成骨分化，维持氧化应激状态下的大鼠BMSCs细胞活性。

关键词: 白藜芦醇, 氧化应激, 成骨分化, 铁死亡

Abstract: Objective:?To explore the protective effect of Res on BMSCs under oxidative stress in vitro. Methods:? Rat BMSCs were treated with 200 μmol/L hydrogen peroxide to undergo oxidative stress. CCK8, ROS fluorescent probe, qRT-PCR, Western blot and ALP staining examined the effects of different concentration of Res on the cell viability, intracellular ROS levels, osteogenic differentiation and ferroptosis in rat BMSCs under oxidative stress. Results:?Res between 1 and 25 μmol/L promoted the cell viability of rat BMSCs stimulated by hydrogen peroxide (P<0.05). 10 μmol/L Res significantly reduced the ROS levels of hydrogen peroxide -stimulated rat BMSCs and increased the expression of antioxidant-related genes, such as NRF2, CAT, SOD-1 and SOD-2 (P＜0.05). 10 μmol/L Res promoted the expression of osteogenic-related markers of hydrogen peroxide -stimulated rat BMSCs (P＜0.05), significantly enhanced the ALP staining effect, promoted the expression of ferroptosis-related markers (GPX4, XCT) (P＜0.05), decreased the expression of acyl-CoA synthetase long-chain family member(ACSL4), prostaglandin-endoperoxide synthase 2 (PTGS2) and nuclear receptor coactivator 4 (NCOA4) (P＜0.05). Conclusions:?Res could reduce the level of ROS and ferroptosis of oxidative stress-induced rat BMSCs, maintain the cell viability and promote the osteogenic differentiation of rat BMSCs under oxidative stress.

Key words: Resveratrol, oxidative stress, osteogenic differentiation, ferroptosis

兰冬梅姚超李雪刘海霞祁胜财. 白藜芦醇通过抑制铁死亡保护骨髓间充质干细胞[J]. 口腔生物医学, 2023, (3): 162-168.

参考文献

[1]. Khosla S, Hofbauer LC. Osteoporosis treatment: recent developments and ongoing challenges. Lancet Diabetes Endocrinol. 2017;5(11):898-907.
[2]. Ebeling PR, Nguyen HH, Aleksova J, et al. Secondary Osteoporosis. Endocr Rev. 2022;43(2):240-313.
[3]. Atashi F, Modarressi A, Pepper MS. The role of reactive oxygen species in mesenchymal stem cell adipogenic and osteogenic differentiation: a review. Stem Cells Dev. 2015;24(10):1150-1163.
[4]. Bartell SM, Kim HN, Ambrogini E, et al. FoxO proteins restrain osteoclastogenesis and bone resorption by attenuating H2O2 accumulation. Nat Commun. 2014;5:3773.
[5]. Deng S, Dai G, Chen S, et al. Dexamethasone induces osteoblast apoptosis through ROS-PI3K/AKT/GSK3beta signaling pathway. Biomed Pharmacother. 2019;110:602-608.
[6]. Yang F, Yang L, Li Y, et al. Melatonin protects bone marrow mesenchymal stem cells against iron overload-induced aberrant differentiation and senescence. J Pineal Res. 2017;63(3).
[7]. Park E, Chung SW. ROS-mediated autophagy increases intracellular iron levels and ferroptosis by ferritin and transferrin receptor regulation. Cell Death Dis. 2019;10(11):822.
[8]. Ma H, Wang X, Zhang W, et al. Melatonin Suppresses Ferroptosis Induced by High Glucose via Activation of the Nrf2/HO-1 Signaling Pathway in Type 2 Diabetic Osteoporosis. Oxid Med Cell Longev. 2020;2020:9067610.
[9]. Galiniak S, Aebisher D, Bartusik-Aebisher D. Health benefits of resveratrol administration. Acta Biochim Pol. 2019;66(1):13-21.
[10]. Zhu SY, Zhuang JS, Wu Q, et al. Advanced oxidation protein products induce pre-osteoblast apoptosis through a nicotinamide adenine dinucleotide phosphate oxidase-dependent, mitogen-activated protein kinases-mediated intrinsic apoptosis pathway. Aging Cell. 2018;17(4):e12764.
[11]. Agidigbi TS, Kim C. Reactive Oxygen Species in Osteoclast Differentiation and Possible Pharmaceutical Targets of ROS-Mediated Osteoclast Diseases. Int J Mol Sci. 2019;20(14).
[12]. Mohsin S, Baniyas MM, AlDarmaki RS, et al. An update on therapies for the treatment of diabetes-induced osteoporosis. Expert Opin Biol Ther. 2019;19(9):937-948.
[13]. Ensrud KE. Bisphosphonates for Postmenopausal Osteoporosis. JAMA. 2021;325(1):96.
[14]. Yang G, Chang CC, Yang Y, et al. Resveratrol Alleviates Rheumatoid Arthritis via Reducing ROS and Inflammation, Inhibiting MAPK Signaling Pathways, and Suppressing Angiogenesis. J Agric Food Chem. 2018;66(49):12953-12960.
[15]. Boissy P, Andersen TL, Abdallah BM, et al. Resveratrol inhibits myeloma cell growth, prevents osteoclast formation, and promotes osteoblast differentiation. Cancer Res. 2005;65(21):9943-9952.