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目的:积雪草是一种传统中医药物,具有很多医学作用,如创伤愈合和抗氧化。中医也发现它可用于白癜风复色和炎症后色素沉着。本研究评估羟基积雪草苷对氧化应激下人黑素细胞作用效果及机制。方法:正常的人黑素细胞予0.01 mmol/L过氧化氢处理,再予不同浓度的羟基积雪草苷(0、10、50、100μg/mL)。评估人黑素细胞树突的回缩速率,流式细胞术及细胞内游离钙浓度([Ca~(2+)]_i)检测线粒体膜电位(MMP),透射电子显微镜(TEM)观察线粒体超微结构,吖啶橙染色检测自噬,蛋白质印迹检测自噬体形成的指标LC3-Ⅱ/LC3-Ⅰ的比值。结果:在过氧化氢引发的氧化应激条件中,羟基积雪草苷在一定浓度下抑制了黑素细胞树突的回缩,提高了膜电位并减少了[Ca~(2+)]_i的积累,减弱了线粒体的损伤,增强了自噬活动。结论:人黑素细胞遭受氧化应激损伤时,羟基积雪草苷可通过自噬激活起到抗氧化作用。因此,羟基积雪草苷可成为氧化应激引起的白癜风的有效治疗手段。
Purpose: Centella is a traditional Chinese medicine that has many medical effects such as wound healing and anti-oxidant. Chinese medicine also found that it can be used for vitiligo reflex and post-inflammatory pigmentation. This study evaluated the effects of asiaticoside on human melanocytes under oxidative stress and its mechanism. Methods: Normal human melanocytes were treated with 0.01 mmol / L H2O2, and then madecassoside at different concentrations (0, 10, 50, 100 μg / mL). Mitochondrial membrane potential (MMP) was measured by flow cytometry and intracellular free calcium concentration ([Ca ~ (2 +)] _i), and mitochondrial ultrastructure was observed by transmission electron microscopy The microstructure and autophagy were detected by acridine orange staining. The ratio of LC3-Ⅱ / LC3-Ⅰ, an indicator of autophagosome formation, was detected by Western blotting. Results: In the hydrogen peroxide-induced oxidative stress, madecassoside inhibited the retraction of melanocytes dendrites at a certain concentration, increased the membrane potential and decreased the [Ca 2+] i Accumulation, weakened mitochondrial damage, increased autophagy activity. CONCLUSION: Madecassoside can play an antioxidant role through activation of autophagy when human melanocytes are damaged by oxidative stress. Therefore, madecassoside can be an effective treatment of vitiligo caused by oxidative stress.