富硒羊肚菌多糖的制备及其体外抗肿瘤活性探究

Preparation of Selenium-enriched Morchella Polysaccharide and Its Antitumor Activity in Vitro

本研究旨在优化提取和纯化富硒羊肚菌多糖(Se-MPS),并评估其抗肿瘤活性及其机制。采用单因素和响应面优化法,对富硒羊肚菌粗多糖的提取工艺进行优化。通过柱层析法对粗多糖进行纯化,并对其化学组成进行分析。使用HepG2细胞进行CCK-8试验、集落形成试验、划痕试验和Transwell试验来评价Se-MPS的抗肿瘤活性。采用RT-qPCR法、Western blot法和免疫荧光法评价Se-MPS的潜在抗肿瘤机制。单因素和响应面试验结果显示,富硒羊肚菌粗多糖提取的最佳工艺参数为:提取时间2.5 h、液料比45:1(mL/g)、提取温度82℃,得率为7.95%。纯化后,Se-MPS的总糖含量达到95.86%,硒含量高达76.07±0.32 mg/g。此外,Se-MPS显著降低了HepG2细胞的集落形成效率、细胞迁移率和侵袭细胞数目(P<0.05)。同时,Se-MPS抑制了PI3K/Akt通路传导,下调了葡萄糖转运蛋白2(GLUT2)的mRNA表达,并降低了细胞的葡萄糖摄取量。研究结果表明,通过对提取时间、液料比及提取温度的优化,可改善富硒羊肚菌粗多糖得率。此外,通过调控PI3K/Akt通路,富硒羊肚菌多糖可发挥出较强的抗肿瘤活性。这项研究为富硒羊肚菌多糖的抗肿瘤活性提供了证据,并为相关功能性食品的开发提供了参考。

This study aimed to optimize the extraction and purification processes for selenium-enriched Morchella esculenta(L.)Pers.(also known as Morchella)polysaccharides(Se-MPS)and to evaluate their anti-tumor activities and mechanisms.The extraction process was optimized using single-factor and response surface methodology.The crude polysaccharides were purified through column chromatography and their chemical composition was analyzed.The anti-tumor activity of Se-MPS was evaluated using HepG2 cells through a series of assays,including CCK-8,colony formation,scratching,and Transwell assays.The potential anti-tumor mechanisms of Se-MPS were assessed using RT-qPCR,Western blot,and immunofluorescence methods.Experimental results from single-factor and response surface methodology revealed that the optimal extraction conditions for selenium-enriched Morchella crude polysaccharides were:An extraction time of 2.5 h,a liquid-to-material ratio of 45:1(mL/g),and an extraction temperature of 82℃,yielding a crude polysaccharides yield of 7.95%.After purification,the total sugar content of Se-MPS reached 95.86%,with a selenium concentration of 76.07±0.32 mg/g.Furthermore,Se-MPS significantly reduced colony formation efficiency,cell migration rate,and the number of invasive cells in HepG2 cells(P<0.05).Meanwhile,Se-MPS inhibited the PI3K/Akt signaling pathway,down-regulated mRNA expression of glucose transporter 2(GLUT2),and reduced glucose uptake in cells.The results indicated that optimizing extraction time,liquid-to-material ratio,and extraction temperature could improve the yield of seleniumenriched Morchella crude polysaccharides.Additionally,Se-MPS exhibited strong anti-tumor activity by regulating the PI3K/Akt pathway.This study provides evidence of the anti-tumor activities of Se-MPS and offers insights for the development of related functional foods.