Lin Mei-Huei, Liu Shyun-Yeu, Liu Young-Chau
Department of Biotechnology, Chia Nan University of Pharmacy and Science, Tainan, Taiwan, ROC.
Autophagy. 2008 Oct;4(7):967-8. doi: 10.4161/auto.6821. Epub 2008 Oct 20.
We recently identified an autophagy-inducing areca nut ingredient (AIAI) in the partially purified 30-100 kDa fraction of areca nut extract (ANE), designated as ANE 30-100K. Before disintegration, most ANE 30-100K-treated cells exhibit rounding morphology, cytoplasmic clearance, and nuclear shrinkage, distinct from arecoline- and cisplatin-induced cellular apoptosis. This unique death pattern is verified to be autophagy by LC3-I cleavage, acidic vesicles, and autophagic vacuoles. As analyzed by Molish's Test, Selinowaff's Test, and thin-layer chromatography, most of the ANE 30-100K constituents are carbohydrates, whereas the protein content of this fraction is less than 1% as assessed by protein assay reagent. The cytotoxicity of ANE 30-100K is further shown to be sensitive to cellulase and proteinase K digestion suggesting AIAI in ANE 30-100K to be a proteoglycan (or glycoprotein). Thus, although ANE contains apoptosis-inducing ingredients such as arecoline, it predominantly triggers autophagic cell death by this natural AIAI.
我们最近在槟榔提取物（ANE）部分纯化的30 - 100 kDa组分中鉴定出一种自噬诱导槟榔成分（AIAI），命名为ANE 30 - 100K。在解体之前，大多数经ANE 30 - 100K处理的细胞呈现圆形形态、细胞质清除和核收缩，这与槟榔碱和顺铂诱导的细胞凋亡不同。通过LC3 - I裂解、酸性囊泡和自噬泡证实这种独特的死亡模式为自噬。经Molish试验、Selinowaff试验和薄层色谱分析，ANE 30 - 100K的大多数成分是碳水化合物，而通过蛋白质测定试剂评估该组分的蛋白质含量小于1%。ANE 30 - 100K的细胞毒性进一步显示对纤维素酶和蛋白酶K消化敏感，表明ANE 30 - 100K中的AIAI是一种蛋白聚糖（或糖蛋白）。因此，尽管ANE含有诸如槟榔碱等诱导凋亡的成分，但它主要通过这种天然的AIAI触发自噬性细胞死亡。
