Department of Spleen-Stomach, Zhenjiang Affiliated Hospital of Nanjing University of Traditional Chinese Medicine, Zhenjiang, China.
Department of Spleen-Stomach, Zhenjiang Hospital of Traditional Chinese Medicine, Zhenjiang, China.
Biomed Res Int. 2019 Jul 25;2019:3103946. doi: 10.1155/2019/3103946. eCollection 2019.
Suitable content of iron is essential for human body, but iron overload is associated with many kinds of diseases including chronic liver damage. Recently, researchers find that iron overload promotes hepatocyte autophagy and apoptosis. However, the mechanism of iron overload in liver damage remains unclear. In this study, Lo2 cells were selected as the research object, iron dextran was a model drug, and astragaloside IV was a therapeutic drug to explore the role of iron overload. MTT assay and Annexin/PI double staining were used to measure cell viability and apoptosis. Ultrastructure was observed by transmission electron microscopy. The expression levels of apoptosis and autophagy-related proteins were determined by real-time PCR and Western Blot. The results showed that iron dextran could significantly inhibit Lo2 cell viability and increase the apoptosis rate, while astragaloside IV could reverse the inhibition of Lo2 cell viability and decrease the apoptosis rate. Transmission electron microscopy showed a significant increase in the number of autophagosomes after administration of iron dextran, and the application of astragaloside IV reduced the production of autophagosomes. LC3II/I was significantly upregulated in the model group but decreased in the astragaloside IV treatment group, and P62 showed the opposite trend. Iron dextran significantly upregulated the expression of Bax and downregulated Bcl2, while astragaloside IV reversed this trend. Finally, the inhibition of hepcidin caused by iron dextran was counteracted by astragaloside IV. In conclusion, the experimental results show that the iron overload model mainly induces excessive autophagy and apoptosis of hepatocytes, thus causing damage to hepatocytes, but astragaloside IV plays a certain therapeutic role in reversing this damage.
适宜的铁含量对人体至关重要，但铁过载与许多疾病有关，包括慢性肝损伤。最近，研究人员发现铁过载可促进肝细胞自噬和凋亡。然而，铁过载导致肝损伤的机制尚不清楚。在本研究中，选择 Lo2 细胞作为研究对象，以右旋糖酐铁为模型药物，以黄芪甲苷 IV 为治疗药物，探讨铁过载的作用。MTT 法和 Annexin/PI 双染法检测细胞活力和凋亡，透射电镜观察超微结构，实时 PCR 和 Western blot 检测凋亡和自噬相关蛋白的表达水平。结果表明，右旋糖酐铁可显著抑制 Lo2 细胞活力，增加细胞凋亡率，而黄芪甲苷 IV 可逆转 Lo2 细胞活力的抑制，降低细胞凋亡率。电镜下观察到铁过载给药后自噬体数量明显增加，而黄芪甲苷 IV 的应用减少了自噬体的产生。LC3II/I 在模型组中明显上调，而在黄芪甲苷 IV 处理组中下调，P62 则呈相反趋势。模型组 Bax 表达明显上调，Bcl2 表达下调，而黄芪甲苷 IV 可逆转这一趋势。最后，黄芪甲苷 IV 可逆转右旋糖酐铁对铁调素的抑制作用。综上所述，实验结果表明，铁过载模型主要诱导肝细胞过度自噬和凋亡，从而导致肝细胞损伤，但黄芪甲苷 IV 在逆转这种损伤方面发挥一定的治疗作用。
