响应面法优化广叶绣球菌蛋白制备工艺及其体外抗消化道肿瘤活性评价  

Optimization for preparation process of the protein from Sparassis latifolia by response surface methodology and evaluation on its anti-gastrointestinal cancer activities in vitro

作　　者：李秀杰 冯彩玲 杨驰[2] 林智 吴建国[1] 吴岩斌[1] LI Xiujie;FENG Cailing;YANG Chi;LIN Zhi;WU Jianguo;WU Yanbin(School of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350012,Fujian,China;Institute of Edible Fungi,Fujian Academy of Agricultural Science,Fuzhou 350012,Fujian,China;Fujian Tianyi Mushroom Industry Co.Ltd.,Youxi 365100,Fujian,China)

机构地区：[1]福建中医药大学药学院,福建福州350012 [2]福建省农业科学院食用菌研究所,福建福州350012 [3]福建天益菌业有限公司,福建尤溪365100

出　　处：《菌物学报》2025年第2期125-137,共13页Mycosystema

基　　金：福建省自然科学基金(2022J01866);国家自然科学基金(81700524)。

摘　　要：本研究制备广叶绣球菌Sparassis latifolia的乙醇提取物(SLEE)、粗多糖(SLCP)和总蛋白(SLTP)并分析各主成分含量,MTT法检测SLEE、SLCP和SLTP对HepG2、MGC-803和HT-293种不同人消化道肿瘤细胞生长的影响。响应面法优化SLTP的制备工艺,测定不同剂量(12.5、25、50、100和200μg/mL)的不同饱和度硫酸铵连续沉淀蛋白对HT-29细胞生长的抑制作用。结果表明SLEE的提取率为(22.75±0.27)%,总三萜含量为(11.40±0.20)%;SLCP的提取率为(7.29±0.23)%,总糖含量为(59.66±0.80)%;SLTP提取率为(8.80±0.57)%,总蛋白含量为(44.59±0.43)%。SLTP对消化道肿瘤细胞的生长抑制作用最强,对HepG2、MGC-803和HT-29细胞的半数抑制浓度IC_(50)值(24 h)分别为44.18、34.27和28.93μg/mL。SLTP最佳提取工艺为:NaCl浓度0.15 mol/L,料液比(g/mL)1:26,浸提时间13 h。在此条件下,SLTP的平均得率为(11.03±0.07)%。30%–50%饱和度的硫酸铵沉淀蛋白(SLSP-50)对HT-29细胞的生长抑制活性最强,半数抑制浓度IC_(50) 值(24 h)为18.84μg/mL。广叶绣球菌蛋白具有显著的体外消化道肿瘤细胞生长抑制活性,是抗肿瘤的主要有效组分之一。本研究为广叶绣球菌资源的充分利用及深加工提供了重要的实验参考依据。The ethanol extract(SLEE),crude polysaccharide(SLCP)and total protein(SLTP)of Sparassis latifolia were prepared and their main constituent content was analyzed.The effects of SLEE,SLCP and SLTP on the growth of three different human gastrointestinal cancer(GI)cell lines,viz.HepG2,MGC-803 and HT-29 cells,were detected by using MTT assay.Response surface methodology was employed to optimize the preparation process of SLTP,and the inhibitory effects of different doses(12.5,25,50,100 and 200μg/mL)of different protein successively precipitated by increasing saturation ammonium sulfate on the growth of HT-29 cells were determined.The results indicated that extraction rate of SLEE was(22.75±0.27)%,with total triterpene content of(11.40±0.20)%;SLCP extraction rate was(7.29±0.23)%,with total sugar content of(59.66±0.80)%;SLTP extraction rate was(8.80±0.57)%,with total protein content of(44.59±0.43)%.SLTP possessed the strongest inhibitory effect on the growth of the GI cancer cell lines,with the IC_(50) values(24 h)of 44.18,34.27 and 28.93μg/mL against HepG2,MGC-803 and HT-29 cells,respectively.The optimal extraction process of SLTP was as follows:the concentration of sodium chloride was 0.15 mol/L,the ratio of solid to liquid was 1:26,and the extraction time was 13 h.Under the optimized conditions,the average yield of SLTP was(11.03±0.07)%.Protein precipitated by 30%–50%saturation ammonium sulfate(SLSP-50)exhibited the most significant inhibitory activity on the growth of HT-29 cells,with the IC_(50) value(24 h)of 18.84μg/mL.The protein of Sparassis latifolia possessed significant inhibitory property on the growth of GI cancer cells in vitro,being one of the main effective ingredients responsible for the anticancer effect.

关 键 词：绣球菌 蛋白 响应面法 工艺优化 抗消化道肿瘤活性 

分 类 号：R284.2[医药卫生—中药学] R285[医药卫生—中医学]