Abstract: This study primarily aimed to explore the optimal protein removal method for polysaccharide of the seed of Livistona chinensis. Using protein clearance rate and polysaccharide retention rate as evaluation indicators, the effects of Sevage method, enzymatic method, and the combination of enzyme and Sevage method on the protein removal from polysaccharide of the seeds were investigated. Additionally, the influence of the obtained polysaccharide of the seeds on RANKL-induced osteoclast differentiation in RAW264.7 cells was explored. The protein removal rates of polysaccharide of the seed using the Sevage method, enzymatic method, and the combined method were 78.85%, 82.40%, and 82.73%, respectively. The polysaccharide retention rates were 72.42%, 92.16%, and 96.72%, respectively. Treatment of RANKL-induced RAW264.7 osteoclast models with polysaccharide of the seed of L. chinensis obtained using the optimal protein removal method revealed that it could reduce the number of osteoclasts generated and decrease their TRAP activity. Therefore, the combination of enzyme and Sevage for protein removal was a relatively effective method that could significantly reduce the protein content in polysaccharide of L. chinensis seed while maintaining a high polysaccharide retention rate. The obtained polysaccharide could also inhibit RANKL-induced osteoclast differentiation in RAW264.7 cells, providing theoretical support for the development of new anti-osteoporosis drugs.

Key words: Livistona chinensis seed, polysaccharide, deproteinization, osteoclast, RANKL

摘要： 以蛋白质清除率和多糖保留率为指标，探讨Sevage法、酶法及两者相结合的方法对蒲葵子多糖脱蛋白的影响，并分析所得多糖对RANKL诱导RAW264.7破骨细胞分化的影响。Sevage法、酶法以及酶与Sevage结合的方法对蒲葵子多糖蛋白的脱除率依次为78.85%、82.40%、82.73%，多糖保留率依次为72.42%、92.16%、96.72%。用最佳的除蛋白方法获得的蒲葵子多糖处理RANKL诱导的RAW264.7破骨细胞模型，发现蒲葵子多糖可以减少破骨细胞生成数量，使得其TRAP活性降低。因此，酶与Sevage结合脱蛋白是相对有效的减少蒲葵子多糖中蛋白含量且多糖保留率较高的方法，所获得的蒲葵子多糖能抑制RANKL诱导RAW264.7向破骨细胞分化，为研发新的抗骨质疏松药物提供理论支撑。

关键词: 蒲葵子, 多糖, 脱蛋白, 破骨细胞, RANKL