Deng Xinran, Zhang Yan, He Xiwei, Li Li, Yue Zhongbin, Liang Yong, Huang Yue
Pengzhou People's Hospital Ophthalmology Department, Chengdu City, 611930, Sichuan Province, 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 Bioenerg Biomembr. 2024 Dec;56(6):619-630. doi: 10.1007/s10863-024-10044-9. Epub 2024 Nov 14.
Oxidative stress-induced lens epithelial cells (LECs) death plays a pivotal role in pathogenesis of age-related cataract (ARC), causing significant visual impairment. Apoptosis of porcine granulosa cells mediated by MMP2 is linked to DNA damage. The current study aimed to investigate the potential mechanism of MMP2 in DNA damage, apoptosis and senescence of lens epithelial cells caused by oxidative stress. HLE-B3 cells were treated with different doses of HO for 24 h, and CCK-8 was used to detect cell viability. Furthermore, western blotting was used to detect the expressions of MMP2, Bcl2, Bax, cleaved caspase3, γ-H2AX, p16, p21, and TIMP2. DCFH-DA staining was used to assess ROS levels. Moreover, EdU staining was used to detect cell proliferation, and flow cytometry was used to detect cell apoptosis. Then, 15A3 immunofluorescence staining and γ-H2AX staining were used to detect DNA damage. In addition, SA-β-gal staining was used to observe cell senescence. The present findings suggest that oxidative stress triggers damage to LECs viability and elevates the expression of MMP2. Furthermore, MMP2 interference attenuates HO-induced active damage, apoptosis, DNA damage, and cellular senescence in LECs. Additionally, TIMP2 expression is down-regulated in HO-induced LECs, which suppresses the expression of MMP2 induced by HO. These findings highlight the crucial role of MMP2 and TIMP2 in the modulation of oxidative stress-induced cellular responses in LECs. Collectively, TIMP2 alleviates HO-induced lens epithelial cell viability damage, apoptosis, DNA damage and cell senescence in LECs by inhibiting MMP2.
氧化应激诱导的晶状体上皮细胞（LECs）死亡在年龄相关性白内障（ARC）的发病机制中起关键作用，可导致严重的视力损害。基质金属蛋白酶2（MMP2）介导的猪颗粒细胞凋亡与DNA损伤有关。本研究旨在探讨MMP2在氧化应激引起的晶状体上皮细胞DNA损伤、凋亡和衰老中的潜在机制。用不同剂量的过氧化氢（HO）处理人晶状体上皮细胞系HLE-B3细胞24小时，并用细胞计数试剂盒-8（CCK-8）检测细胞活力。此外，采用蛋白质免疫印迹法检测MMP2、Bcl2、Bax、裂解的半胱天冬酶3、γ-H2AX、p16、p21和组织金属蛋白酶抑制剂2（TIMP2）的表达。用2',7'-二氯二氢荧光素二乙酸酯（DCFH-DA）染色评估活性氧（ROS）水平。此外，用5-乙炔基-2'-脱氧尿苷（EdU）染色检测细胞增殖，用流式细胞术检测细胞凋亡。然后，用15A3免疫荧光染色和γ-H2AX染色检测DNA损伤。此外，用衰老相关β-半乳糖苷酶（SA-β-gal）染色观察细胞衰老。本研究结果表明，氧化应激触发对LECs活力的损伤并提高MMP2的表达。此外，MMP2干扰减弱HO诱导的LECs活性损伤、凋亡、DNA损伤和细胞衰老。另外，在HO诱导的LECs中TIMP2表达下调，这抑制了HO诱导的MMP2表达。这些发现突出了MMP2和TIMP2在调节氧化应激诱导的LECs细胞反应中的关键作用。总的来说，TIMP2通过抑制MMP2减轻HO诱导的LECs活力损伤、凋亡、DNA损伤和细胞衰老。
