一氧化氮通过VDAC1调控钙稳态诱导人原代黑素细胞凋亡。

IF 3.7 2区 生物学 Q3 CELL BIOLOGY
Jiawei Lu, Yifei Feng, Yidan Wang, Yongkai Yu, Wene Zhao, Xuechen Cao, Ziyu Li, Yan Lu
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引用次数: 0

摘要

白癜风是一种自身免疫性皮肤病,起源于黑色素细胞(MCs)的损伤和损失。研究发现白癜风患者一氧化氮(NO)代谢异常,但NO导致MC丢失的机制尚不清楚。收集白癜风患者和健康对照者的皮肤样本,评估三种一氧化氮合酶(NOS)亚型的表达。将角质形成细胞、MCs和成纤维细胞暴露于细胞因子混合物(IFN-γ、TNF-α、IL-1β和LPS)中,以模拟白癜风的促炎微环境。然后检测各细胞类型的NOS激活和NO生成能力。以硝普钠(SNP)作为NO供体,评价其对MCs的细胞毒作用。在SNP暴露后,测量细胞活力、台盼蓝染色率和乳酸脱氢酶释放。透射电镜观察MCs的超微结构变化。流式细胞术检测细胞凋亡率、细胞内钙浓度和线粒体膜电位。利用BAPTA-AM进行细胞内钙螯合,利用小干扰RNA沉默MCs中VDAC1的表达。诱导型NOS (iNOS)在白癜风活动性病变中表达显著上调。在促炎条件下,角质形成细胞和成纤维细胞通过iNOS激活产生增加的NO。暴露于高no环境的MCs表现出剂量依赖性的细胞损伤。snp处理的MCs表现出凋亡的超微结构特征,包括核染色质浓缩和线粒体肿胀。流式细胞术和促凋亡标志物分析证实了线粒体介导的MCs凋亡。钙螯合和VDAC1沉默均可通过恢复钙稳态和线粒体膜电位来减轻MCs的凋亡。no诱导的MCs凋亡是由钙超载和线粒体功能障碍介导的。iNOS激活可能是白癜风中黑色素细胞丢失的一个因素,也是一个潜在的治疗靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Nitric oxide induces apoptosis of human primary melanocytes by regulating calcium homeostasis via VDAC1.

Vitiligo is an autoimmune skin disease that originates from damage and loss of melanocytes (MCs). Studies have identified abnormal nitric oxide (NO) metabolism in vitiligo patients, but the mechanisms by which NO contributes to MC loss remain unclear. Skin samples from patients with vitiligo and healthy controls were collected to evaluate the expression of three nitric oxide synthases (NOS) isoforms. Keratinocytes, MCs, and fibroblasts were exposed to a cytokine cocktail (IFN-γ, TNF-α, IL-1β and LPS) to simulate the pro-inflammatory microenvironment of vitiligo. Then NOS activation and the capacity for NO production were examined in each cell type. Sodium nitroprusside (SNP) was used as the NO donor to evaluate its cytotoxic effects on MCs. Cell viability, trypan blue staining rate, and lactate dehydrogenase release were measured following SNP exposure. Ultrastructural changes in MCs were observed using transmission electron microscopy. Apoptosis rate, intracellular calcium concentration, and mitochondrial membrane potential were assessed using flow cytometry. BAPTA-AM was used for intracellular calcium chelation and small interfering RNA was used to silence VDAC1 expression in MCs. Inducible NOS (iNOS) expression was significantly upregulated in patients in the active vitiligo lesions. Under pro-inflammatory conditions, keratinocytes and fibroblasts produced increased NO via iNOS activation. MCs exposed to a high-NO environment displayed dose-dependent cellular damage. SNP-treated MCs showed ultrastructural features of apoptosis, including condensed nuclear chromatin and swollen mitochondria. Flow cytometry and analysis of pro-apoptotic markers confirmed mitochondrial-mediated apoptosis in MCs. Both calcium chelation and VDAC1 silencing alleviated the apoptosis of MCs by restoring calcium homeostasis and mitochondrial membrane potential. NO-induced apoptosis in MCs was mediated by calcium overload and mitochondrial dysfunction. iNOS activation may represent a contributor to melanocyte loss in vitiligo and a potential therapeutic target.

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来源期刊
Molecular and Cellular Biochemistry
Molecular and Cellular Biochemistry 生物-细胞生物学
CiteScore
8.30
自引率
2.30%
发文量
293
审稿时长
1.7 months
期刊介绍: Molecular and Cellular Biochemistry: An International Journal for Chemical Biology in Health and Disease publishes original research papers and short communications in all areas of the biochemical sciences, emphasizing novel findings relevant to the biochemical basis of cellular function and disease processes, as well as the mechanics of action of hormones and chemical agents. Coverage includes membrane transport, receptor mechanism, immune response, secretory processes, and cytoskeletal function, as well as biochemical structure-function relationships in the cell. In addition to the reports of original research, the journal publishes state of the art reviews. Specific subjects covered by Molecular and Cellular Biochemistry include cellular metabolism, cellular pathophysiology, enzymology, ion transport, lipid biochemistry, membrane biochemistry, molecular biology, nuclear structure and function, and protein chemistry.
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