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[摘要]
探讨了蓝莓黑醋栗枸杞决明子复合物(Blueberry,Blackcurrant,Medlar,Cassia Seed Complex,LHGJ)对丙酮醛(Methylglyoxal,MGO)诱导的人视网膜上皮细胞(Retinal Pigment Epithelium,APRE-19)氧化应激损伤的保护作用。以MGO诱导ARPE-19细胞氧化应激损伤建立细胞模型,给予不同质量浓度LHGJ复合物处理。利用CCK-8法检测细胞活力,采用荧光显微镜检测细胞活性氧(Reactive Oxygen Species,ROS)含量、采用酶联免疫法(Enzyme-linked Immunosorbent Assay,ELISE)测定一氧化氮(Nitric Oxide,NO)含量、比色法测定谷胱甘肽过氧化物酶(Glutathione Peroxidase,GSH-Px)及羟胺法测定超氧化物歧化酶(Superoxide Dismutase,SOD)活性。实验结果表明,ARPE-19细胞可经0.80 mmol/L MGO诱导氧化应激损伤模型。诱导ARPE-19细胞经0.10、1.00 μg/mL LHGJ复合物处理后,细胞活力增加23.03%与21.23%,具有高度显著差异(P<0.001)。0.01、0.10、10.00 μg/mL LHGJ复合物使ROS的荧光强度显著降低1.02、1.28、2.37倍(P<0.05),0.01、0.10、10 μg/mL LHGJ复合物使NO的生成量显著降低13.24、15.23、16.24 μmol/g prot(P<0.05);1.00、10.00 μg/mL LHGJ 复合物使SOD活性显著与极显著升高6.01、7.43 U/mg(P<0.05、P<0.01);1.00与10.00 μg/mL LHGJ复合物显著增加了GSH-Px活力14.32及25.42 U/mg(P<0.05)。LHGJ 复合物对MGO导致的ARPE-19细胞氧化应激损伤具有保护作用,可为LHGJ进一步开发提供理论依据。
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[Abstract]
To study the protective effect of blueberry, blackcurrant, medlar, cassia seed complex (LHGJ) against the oxidative stress injury in human retinal epithelial cells (APRE-19) induced by methylglyoxal (MGO). The ARPE-19 cell model was established through inducing oxidative stress by MGO, and LHGJ solutions at different concentrations were used to treat the cells. Cell viability was determined by the CCK-8 method, the reactive oxygen species (ROS) content in cells was measured by the fluorescence microscopy method, the nitric oxide (NO) content was measured by enzyme-linked immunosorbent assay (ELISA), the glutathione peroxidase (GSH-PX) content was measured by colorimetry, and the superoxide dismutase (SOD) activity was measured by the hydroxylamine method. The results showed that oxidative stress injury in ARPE-19 cells could be induced by 0.80 mmol/L MGO. The induced ARPE-19 cells were then treated with 0.10 or 1.00 μg/mL LHGJ complex, which increased the cell viability by 23.03% and 21.23%, respectively (P<0.001). LHGJ complex solutions at 0.01, 0.10 and 10.00 μg/mL significantly (P<0.05) reduced the fluorescence intensity of ROS (by 1.02, 1.28 and 2.37 times, respectively), LHGJ complex solutions at 0.01, 0.10 and 10.00 μg/mL significantly (P<0.05) decreased NO production (by 13.24, 15.23 and 16.24 μmol/g prot., respectively), LHGJ complex solutions at 1.0 and 10.00 μg/mL significantly (P<0.05) increased the activity of SOD (by 6.01 and 7.43 U/mg, respectively) (P<0.05, P<0.01) and the content of GSH-Px (by 14.32 and 25.42 U/mg, respectively) (P<0.05). LHGJ complex can protect ARPE-19 cells against oxidative stress injury induced by MGO, which provides a theoretical basis for further development of LHGJ complex.
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