Changchun University of Chinese Medicine, Changchun, 130117, China.
Department of Urology, The Affiliated Hospital of Changchun University of Chinese Medicine, Changchun, 130021, China.
Free Radic Biol Med. 2024 Nov 1;224:366-381. doi: 10.1016/j.freeradbiomed.2024.08.043. Epub 2024 Sep 2.
Exploring the therapeutic effect and mechanism of isorhamnetin in the treatment of DMED.
Using a high glucose environment to induce endothelial cells damage in the corpus cavernosum, and combining with intervention agents such as ferroptosis inhibitors to observe the process of cell damage and repair, evaluating cell status through CCK-8 and DAPI; To establish the STZ-induced diabetes rat model and detect the erectile function and tissue changes; Perform transcriptomic sequencing on rat models and samples treated with isorhamnetin to analyze differentially expressed genes and their GO functions; Identify critical pathways by combining with the ferroptosis database; Flow cytometry was used to detect ROS and mitochondrial membrane potential, and RT-PCR was used to verify gene expression, Seahorse detects mitochondrial oxygen consumption rate, revealing the mechanism of action of isorhamnetin.
Ferroptosis inhibitors and isorhamnetin can effectively reverse the damage of corpus cavernosum endothelial cells induced by high glucose and ferroptosis agonists. Isorhamnetin has the ability to reinstate the erectile function of diabetic rats, while enhancing the quantity of endothelial cells and refining the morphology of collagen fibers. Immunohistochemistry revealed that ferroptosis existed in the penis tissue of diabetes rats. Transcriptomic analysis showed that isorhamnetin improves gene expression in DM rats by regulating genes such as GFER, IGHM, GPX4 and HMOX1, involving multiple pathways and biological processes. Flow cytometry and RT-PCR confirmed that isorhamnetin can reduce reactive oxygen species levels, restore essential gene expression, improve mitochondrial membrane potential, and alleviate oxidative stress and ferroptosis. Seahorse detection found that isorhamnetin can restore mitochondrial oxygen consumption rate.
Isorhamnetin attenuates high glucose damage to cavernous endothelial cells by inhibiting ferroptosis and oxidative stress, restores erectile function and improves tissue morphology in diabetic rats, and its multi-pathway and multi-targeting regulatory mechanism suggests that it is promising to be an effective drug for the treatment of DMED.
探讨山奈酚治疗 DMED 的疗效及机制。
采用高糖环境诱导海绵体血管内皮细胞损伤，结合铁死亡抑制剂等干预剂观察细胞损伤与修复过程，通过 CCK-8 和 DAPI 评估细胞状态；建立 STZ 诱导的糖尿病大鼠模型，检测勃起功能及组织变化；对山奈酚处理的大鼠模型及样本进行转录组测序，分析差异表达基因及其 GO 功能；结合铁死亡数据库识别关键通路；通过流式细胞术检测 ROS 和线粒体膜电位，RT-PCR 验证基因表达，通过 Seahorse 检测线粒体耗氧率，揭示山奈酚的作用机制。
铁死亡抑制剂和山奈酚可有效逆转高糖和铁死亡激动剂诱导的海绵体血管内皮细胞损伤。山奈酚具有恢复糖尿病大鼠勃起功能的能力，同时增加内皮细胞数量，改善胶原纤维形态。免疫组化显示糖尿病大鼠阴茎组织存在铁死亡。转录组分析表明，山奈酚通过调节 GFER、IGHM、GPX4 和 HMOX1 等基因改善 DM 大鼠的基因表达，涉及多个通路和生物学过程。流式细胞术和 RT-PCR 证实，山奈酚可降低活性氧水平，恢复关键基因表达，改善线粒体膜电位，减轻氧化应激和铁死亡。Seahorse 检测发现，山奈酚可恢复线粒体耗氧率。
山奈酚通过抑制铁死亡和氧化应激减轻高糖对海绵体血管内皮细胞的损伤，恢复糖尿病大鼠的勃起功能，改善组织形态，其多通路、多靶点的调控机制提示其有望成为治疗 DMED 的有效药物。
