Liu Junyi, Tang Yongying, Li Jiang, Zhang Hong, Zha Xu, Chen Quan, Li Jinghua, Zhao Xueying
Department of Ophthalmology, The Second Affiliated Hospital of Kunming Medical University, Kunming, 650000, China.
Department of Ophthalmology, Kunming Children's Hospital, Kunming, 650034, China.
Cell Biochem Biophys. 2025 Mar;83(1):391-402. doi: 10.1007/s12013-024-01470-y. Epub 2024 Aug 8.
Cataracts are characterized as a disease affecting lens opacity. Endoplasmic reticulum (ER) stress can cause lens epithelial cell (LEC) dysfunction, affecting normal lens transparency and function, but the role of Tribbles 3 (TRB3), an inducible gene of ER stress, in cataracts is poorly understood. This study explored how TRB3 promotes cataract progression through ER stress. We administered a subcutaneous injection of sodium selenite at a dosage of 3.46 mg/kg to rats to create an animal model of cataracts. Additionally, we exposed rat LEC cells to 0.01 μM tunicamycin (TM) for 24 h to establish a cell model of ER stress. The detection of related genes and proteins was performed via RT‒qPCR and Western blot techniques. Flow cytometry, along with JC-1, TUNEL, and HE staining, was employed to assess damage to cells and lens tissues. This study revealed that TRB3 was abnormally highly expressed in both a cataract rat model and an ER stress cell model. Knocking down TRB3 has a similar effect as treatment with an ER stress inhibitor, effectively reversing the ER stress and apoptosis induced by TM. This effect includes increasing the mitochondrial membrane potential in LEC cells, lowering reactive oxygen species (ROS) levels, increasing ATP production, suppressing the expression of the apoptosis-related proteins Bax and C-caspase-3, increasing Bcl-2 expression, and decreasing apoptosis. Furthermore, TRB3 knockdown improved the pathological conditions of rat lenses and inhibited mitochondrial dysfunction and cell apoptosis to relieve the development of cataracts in rats. Mechanistically, CHOP promotes the expression of TRB3 by binding to the TRB3 promoter, thereby activating ER stress, leading to mitochondrial dysfunction and cell apoptosis in LEC cells and accelerating the development of cataracts. According to our findings, targeting TRB3 expression inhibition could emerge as a novel approach for cataract therapy.
白内障的特征是一种影响晶状体透明度的疾病。内质网(ER)应激可导致晶状体上皮细胞(LEC)功能障碍,影响正常晶状体的透明度和功能,但内质网应激诱导基因Tribbles 3(TRB3)在白内障中的作用尚不清楚。本研究探讨了TRB3如何通过内质网应激促进白内障进展。我们以3.46mg/kg的剂量给大鼠皮下注射亚硒酸钠,以建立白内障动物模型。此外,我们将大鼠LEC细胞暴露于0.01μM衣霉素(TM)中24小时,以建立内质网应激细胞模型。通过RT-qPCR和蛋白质免疫印迹技术检测相关基因和蛋白质。采用流式细胞术以及JC-1、TUNEL和苏木精-伊红(HE)染色来评估细胞和晶状体组织的损伤。本研究表明,TRB3在白内障大鼠模型和内质网应激细胞模型中均异常高表达。敲低TRB3具有与内质网应激抑制剂治疗类似的效果,可有效逆转TM诱导的内质网应激和细胞凋亡。这种作用包括增加LEC细胞中的线粒体膜电位、降低活性氧(ROS)水平、增加ATP生成、抑制凋亡相关蛋白Bax和C-半胱天冬酶-3的表达、增加Bcl-2表达以及减少细胞凋亡。此外,敲低TRB3改善了大鼠晶状体的病理状况,抑制了线粒体功能障碍和细胞凋亡,从而缓解了大鼠白内障的发展。机制上,C/EBP同源蛋白(CHOP)通过与TRB3启动子结合促进TRB3的表达,从而激活内质网应激,导致LEC细胞中的线粒体功能障碍和细胞凋亡,并加速白内障的发展。根据我们的研究结果,靶向抑制TRB3表达可能成为一种新的白内障治疗方法。