蓝莓黑醋栗枸杞决明子复合物对眼部细胞氧化应激损伤的保护作用

Protective Effect of a Complex of Blueberry, Blackcurrant, Medlar and Cassia Obtusifolia against Oxidative Stress Injury in Retinal Epithelial Cells

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。

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.