Yang Jun, Gao Yichen, Mao Han, Kuang Xinqi, Tian Fang
Tianjin Key Laboratory of Retinal Functions and Diseases, Tianjin Branch of National Clinical Research Center for Ocular Disease, Eye Institute and School of Optometry, Tianjin Medical University Eye Hospital, Tianjin, 300384, China.
Tianjin Key Laboratory of Retinal Functions and Diseases, Tianjin Branch of National Clinical Research Center for Ocular Disease, Eye Institute and School of Optometry, Tianjin Medical University Eye Hospital, Tianjin, 300384, China.
J Ethnopharmacol. 2024 Oct 28;333:118444. doi: 10.1016/j.jep.2024.118444. Epub 2024 Jun 6.
ETHNOPHARMACOLOGICAL RELEVANCE: Qiju Dihuang Pill (QDP) is a traditional Chinese medicine prescription for the treatment of eye diseases. Novel literature reports that copper-induced cell death, called as cuproptosis, is a copper-dependent and differs distinctly from other types of cell death. AIM OF THE STUDY: The present study aims to investigate whether QDP could protect lens epithelial cells via alleviating copper-induced death in diabetic cataract. MATERIALS AND METHODS: The different concentration of QDP medicated serum was administrated on high glucose (HG)-induced human lens epithelial cells (HLECs). The copper concentration was tested using Elabscience Copper Assay kit. The proliferation was detected using CCK-8 and EdU assays. The molecular binding was identified using RIP-PCR and luciferase reporter assay. RESULTS: Results indicated that HG culture condition triggered the copper concentration and repressed the proliferation of HLECs. Then, the elesclomol-Cu (Es-Cu) administration up-regulated the copper concentration and inhibited the proliferation, and cuproptosis inhibitor tetrathiomolybdate (TTM) could specifically reverse the consequence. QDP treatment reduced the copper concentration and cuproptosis-related genes (SLC31A1, FDX1). MeRIP-Seq and RIP-PCR confirmed that QDP reduced the stability of SLC31A1 mRNA through mA modified site, and copper actually synergized the molecular binding efficiency. Rescue assay verified the role of QDP and SLC31A1 on HLECs' cuproptosis characteristic. CONCLUSION: This research identified the protective role of QDP on HG-induced HLECs in DC through decreasing mA/SLC31A1-mediated cuproptosis in DC. This finding provides novel insights into mechanisms for QDP and sheds light on the multifaceted role of traditional prescription on DC.
民族药理学相关性:杞菊地黄丸(QDP)是一种用于治疗眼部疾病的中药方剂。新的文献报道,铜诱导的细胞死亡,即铜死亡,是一种依赖铜的细胞死亡方式,与其他类型的细胞死亡有明显区别。 研究目的:本研究旨在探讨QDP是否能通过减轻糖尿病性白内障中铜诱导的细胞死亡来保护晶状体上皮细胞。 材料与方法:将不同浓度的QDP含药血清作用于高糖(HG)诱导的人晶状体上皮细胞(HLECs)。使用Elabscience铜检测试剂盒检测铜浓度。采用CCK-8和EdU检测法检测细胞增殖。通过RIP-PCR和荧光素酶报告基因检测法鉴定分子结合情况。 结果:结果表明,HG培养条件会引发铜浓度升高并抑制HLECs的增殖。然后,依拉莫司汀-铜(Es-Cu)处理上调了铜浓度并抑制了增殖,而铜死亡抑制剂四硫代钼酸盐(TTM)可以特异性地逆转这一结果。QDP处理降低了铜浓度以及与铜死亡相关的基因(SLC31A1、FDX1)。MeRIP-Seq和RIP-PCR证实,QDP通过mA修饰位点降低了SLC31A1 mRNA的稳定性,并且铜实际上增强了分子结合效率。挽救实验验证了QDP和SLC31A1对HLECs铜死亡特征的作用。 结论:本研究确定了QDP通过降低糖尿病性白内障中mA/SLC31A1介导的铜死亡对HG诱导的HLECs具有保护作用。这一发现为QDP的作用机制提供了新的见解,并揭示了传统方剂在糖尿病性白内障中的多方面作用。