Li Xiaosha, Tang Shiyang, Wang Haizhen, Li Xin
Department of Dermatology, The Second Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, China.
School of Chinese Medicine, Hunan University of Chinese Medicine, Changsha, China.
Pharmacology. 2025;110(3):141-150. doi: 10.1159/000541177. Epub 2024 Oct 16.
Melanocyte ferroptosis has been proven to contribute to the development of vitiligo. Tanshinone IIA (TSA), a Chinese herbal extract, has been shown to inhibit vitiligo progression. Whether TSA regulates ferroptosis in melanocytes remains unclear.
Hydrogen peroxide (H2O2) was used to induce melanocytes to stimulate vitiligo cell model in vitro. Cell proliferation was examined by 5-ethynyl-2'-deoxyuridine assay. The levels of malondialdehyde, reactive oxygen species, glutathione peroxidase, and iron were detected by corresponding commercial kit. The protein levels of ferroptosis-related markers and Nrf2 pathway-related markers were examined using western blot and immunofluorescence staining. Cell viability and cytotoxicity were analyzed using Cell Counting Kit-8 assay and lactate dehydrogenase detection. Mitochondrial morphology was examined using a transmission electron microscope.
After H2O2 treatment, melanocyte proliferation was reduced, while oxidative stress and ferroptosis were enhanced. TSA treatment could inhibit ferroptosis in H2O2-induced melanocytes. Besides, TSA could activate Nrf2 pathway and promote Nrf2 nuclear translocation, and Nrf2-specific inhibitor (ML385) also reversed the inhibitory effect of TSA on H2O2-induced melanocyte ferroptosis.
Our data showed that TSA alleviated H2O2-induced melanocyte ferroptosis via activating Nrf2 pathway.
黑素细胞铁死亡已被证明与白癜风的发展有关。丹参酮IIA(TSA)是一种中草药提取物,已被证明可抑制白癜风的进展。TSA是否调节黑素细胞中的铁死亡尚不清楚。
用过氧化氢(H2O2)诱导黑素细胞,在体外建立白癜风细胞模型。通过5-乙炔基-2'-脱氧尿苷检测法检测细胞增殖。用相应的商业试剂盒检测丙二醛、活性氧、谷胱甘肽过氧化物酶和铁的水平。使用蛋白质免疫印迹法和免疫荧光染色法检测铁死亡相关标志物和Nrf2通路相关标志物的蛋白水平。使用细胞计数试剂盒-8检测法和乳酸脱氢酶检测法分析细胞活力和细胞毒性。用透射电子显微镜观察线粒体形态。
H2O2处理后,黑素细胞增殖减少,而氧化应激和铁死亡增强。TSA处理可抑制H2O2诱导的黑素细胞铁死亡。此外,TSA可激活Nrf2通路并促进Nrf2核转位,Nrf2特异性抑制剂(ML385)也可逆转TSA对H2O2诱导的黑素细胞铁死亡的抑制作用。
我们的数据表明,TSA通过激活Nrf2通路减轻H2O2诱导的黑素细胞铁死亡。