马氏珠母贝黏液糖蛋白组成及其体外抗癌活性研究

Study on the Composition and Anti-cancer Activity in Vitro of Mucinous Glycoproteins of Pinctada martensii

采用透析、醇沉法提取马氏珠母贝(Pinctada martensii)黏液中的糖蛋白,经Sephadex G-150分离纯化后得糖蛋白纯化品,对糖蛋白纯化品MP-FI、MP-FII进行分子量分布、氨基酸组成及傅里叶红外光谱(FTIR)分析,进一步采用体外细胞试验初步评估MP-FI和MP-FII的体外抗癌生物活性。结果表明,MP-FI和MP-FII均含有与抗癌功能活性相关的赖氨酸和精氨酸且含量较高,MP-FI中赖氨酸和精氨酸的含量占水解氨基酸总量的8.81%和5.80%。MP-FI的分子量集中在6.7~39.0 kDa范围。MP-FI、MP-FII的FTIR特征吸收峰表明2种样品结构组成存在差异,MP-FI具有α螺旋的二级结构和β-型糖苷键的糖链结构。MP-FI、MP-FII均表现出一定的抗癌活性,当MP-FI作用质量浓度达6 mg/mL时,对HepG2的抑制率高达61.56%±0.99%,IC_(50)值为2.7 mg/mL,对LLC的抑制率达42.23%±3.27%,IC_(50)值为12.1 mg/mL,且MP-FI对HepG2、LLC的抑制作用明显高于MP-FII(p<0.05)。研究结果可为探索马氏珠母贝黏液中的生物活性物质提供理论基础数据。

Crude glycoproteins were obtained from the mucus of Pinctada martensii through dialysis and alcohol precipitation.And the mucus of Pinctada martensii glycoproteins were further purified by Sephadex G-150 column chromatography.The molecular weight distribution,amino acid composition and FTIR spectra of the purified glycoprotein MP-FI and MP-FII were analyzed,and the anti-cancer activity in vitro of MP-FI and MP-FII was evaluated by using cell experiment in vitro.The results showed that both MP-FI and MP-FII contain lysine and arginine,which were related to the anti-cancer activity.The contents of lysine and arginine in MP-FI account for 8.81%and 5.80%of the total hydrolyzed amino acids.The molecular weight distribution of MP-FI was in the 6.7~39.0 kDa range.The infrared spectral characteristics of MP-FI and MP-FII indicated that the structure and composition of the two samples were different,MP-FI has the secondary structure ofα-helix and the sugar chain structure ofβ-glycosidic bond.Both MP-FI and MP-FII have certain anticancer activity,and the concentration of MP-FI at 6 mg/mL,the inhibitory rate against HepG2 was as high as 61.56%±0.99%,the IC_(50) was 2.7 mg/mL,meanwhile the inhibitory rate against LLC was as high as 42.23%±3.27%,the IC_(50) was 12.1 mg/mL,respectively.And the inhibitory effect of MP-FI on HepG2 and LLC was significantly higher than that of MP-FII(p<0.05).The results could provide theoretical basis for the exploration of bioactive substances in the mucus of Pinctada martensii.