In this study, a novel water soluble polysaccharide (named GFP-4) was extracted from Grifola frondosa at 4 ^oC, and its preliminary structure and inhibitory effects on human gastric carcinoma MKN-45 cells through the Fas/FasL death receptor apoptosis pathway were investigated. High-performance gel permeation chromatography (HPGPC), fourier-transform infrared spectroscopy (FT-IR), and ion chromatography (IC) results showed that GFP-4 was a 1.09 × 10⁶ Da neutral hetero polysaccharide with pyranose rings, and α- and β-type glycosidic linkages that contained galactose, glucose, and mannose at a molar ratio of 1.00:3.45:1.19. MTT results indicated that GFP-4 significantly inhibited the proliferation of MKN-45 cells in a concentration-dependent manner. The H&E staining and Hoechst 33342/PI double staining results showed that GFP-4-treated MKN-45 cells were subjected to underwent typical apoptotic morphologic changes such as nuclear pyknosis, chromatin condensation, and an increase of membrane permeability. Annexin V-FITC/PI double staining, cell cycle analysis, and western blot results revealed the GFP-4 induced MKN-45 cells apoptosis through the Fas/FasL-mediated death receptor pathway with cells arrested at the G0/G1 phase. These data indicate that GFP-4 is a promising candidate for treating gastric cancer and provide a theoretical basis for the future development and utilization of G. frondosa clinically.

中文翻译：

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灰树花冷水溶性多糖的制备及其对MKN-45细胞的抑制作用。

在这项研究中，一种新型的水溶性多糖（命名为GFP-4）从萃取灰树花在4 ^Ô C，以及它的初步结构，并通过中Fas / FasL的死亡受体凋亡途径对人胃癌MKN-45细胞的抑制效果都调查。高性能凝胶渗透色谱（HPGPC），傅立叶变换红外光谱（FT-IR）和离子色谱（IC）结果显示GFP-4为1.09×10 ⁶具有吡喃糖环的大中性杂多糖，以及包含半乳糖，葡萄糖和甘露糖的摩尔比为1.00：3.45：1.19的α和β型糖苷键。MTT结果表明，GFP-4以浓度依赖性方式显着抑制MKN-45细胞的增殖。H＆E染色和Hoechst 33342 / PI双重染色结果表明，经GFP-4处理的MKN-45细胞经历了典型的凋亡形态变化，例如核固缩，染色质浓缩和膜通透性增加。Annexin V-FITC / PI双重染色，细胞周期分析和蛋白质印迹结果显示，GFP-4通过Fas / FasL介导的死亡受体途径诱导MKN-45细胞凋亡，而细胞停滞在G0 / G1期。G. frondosa临床上。