纳豆激酶对高糖缺氧诱导的视网膜色素上皮细胞损伤的保护作用

目的 探讨纳豆激酶(NK)对高糖缺氧导致的人视网膜色素上皮细胞(ARPE-19)损伤的保护作用。方法 不同浓度的葡萄糖和氯化钴(CoCl₂)诱导培养ARPE-19细胞24 h后，观察细胞形态变化，采用CCK-8法检测ARPE-19细胞活性。Western blot和RT-qPCR检测紧密连接蛋白-1(ZO-1)、缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子A(VEGFA)、白细胞介素(IL)-1β、IL-18的表达情况，筛选最佳葡萄糖和CoCl₂浓度构建体外高糖缺氧的细胞模型。CCK-8法筛选合适的NK浓度，将ARPE-19细胞分为正常糖组(5.5 mmol·L^-1葡萄糖，NC组)、高糖+CoCl₂组(25.0 mmol·L^-1葡萄糖+200μmol·L^-1 CoCl₂,HG+CoCl₂组)和NK处理组(1.0μmol·L^-1 NK+25.0 mmol·L^-1葡...

Protective effect of nattokinase on retinal pigment epithelial cell injury induced by high glucose or hypoxia

Objective　To investigate the protective effect of nattokinase (NK) on human retinal pigment epithelial cell (ARPE-19) injury induced by high glucose or hypoxia. Methods　ARPE-19 cells were cultured with different concentrations of glucose and cobalt chloride (CoCl₂) for 24 hours, then cell morphological changes were observed and cell viability was detected by CCK-8. The expression levels of zonula occludens-1 (ZO-1), hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor A (VEGFA), interleukin (IL)-1β, and IL-18 were measured by Western blot and real-time quantitative polymerase chain reaction (RT-qPCR), respectively, and the optimal glucose and CoCl₂ concentrations were screened to construct the high glucose or hypoxia induced cell models in vitro. The appropriate NK concentration was determined by CCK-8. ARPE-19 cells were divided into the normal control group (5.5 mmol·L^-1 glucose, NC group), high glucose and CoCl₂ group (25.0 mmol·L^-1 glucose + 200 μmol·L^-1 CoCl₂, HG+CoCl₂ group), and NK treatment group (1.0 μmol·L^-1 NK + 25.0 mmol·L^-1 glucose + 200 μmol·L^-1 CoCl₂, NK+HG+CoCl₂ group). The protein and mRNA expression levels of ZO-1, HIF-1α and VEGFA in these groups were detected by Western blot and RT-qPCR. Results　CCK-8 assay results showed that compared with the NC group, the viability of ARPE-19 cells in the intervention groups with CoCl₂ concentration higher than 200 μmol·L^-1 was significantly decreased (both P<0.01), and the cell morphology was significantly changed. Western blot and RT-qPCR results showed that compared with the NC group, the protein and mRNA expression levels of HIF-1α and VEGFA, as well as the mRNA expression levels of IL-1β and IL-18 in the intervention groups (200 μmol·L^-1 CoCl₂) were significantly increased (all P<0.01), while the protein expression level of ZO-1 was significantly decreased (all P<0.05). CCK-8 assay results showed that when NK concentration was higher than 1.0 μmol·L^-1, the viability of ARPE-19 cells was significantly decreased compared with the NC group (all P<0.01). Compared with the HG+CoCl₂ group, the protein and mRNA expression levels of HIF-1α and VEGFA in the NK+HG+CoCl₂ group were significantly decreased (all P<0.001), while the protein expression level of ZO-1 was significantly increased (P<0.05). Conclusion　NK can reduce the protein and mRNA expression levels of HIF-1α and VEGFA in ARPE-19 cells damaged by high glucose or hypoxia, thus protecting these cells.