小刺猴头菌发酵浸膏多糖体外抗氧化活性分析

In vitro antioxidant activity of polysaccharides extracted from Hericium caput-medusae (Bull.: Fr.) Pers

关键词:  小刺猴头菌  多糖  清除自由基  RAW264.7巨噬细胞  抗氧化酶

Key words:  Hericium caput-medusae (Bull.:Fr) Pers.  polysaccharide  free radical scavenging  RAW 264.7 macrophage  antioxidase

摘要:

为研究小刺猴头菌[Hericium caput-medusae（Bull.：Fr.） Pers]发酵浸膏多糖的抗氧化活性。以工厂化小刺猴头菌发酵浸膏为原料，采用透析法分级，DEAE sepharose fast flow柱层析纯化，分别获得粗多糖组分HFCP1和中性多糖HFCP1-1；通过PMP柱前衍生-高效液相色谱法和红外光谱法对单糖组成和结构进行分析，单糖组成均以葡萄糖和半乳糖为主。化学法检测多糖抗氧化活性，结果显示：HFCP1和HFCP1-1清除羟基自由基、超氧阴离子自由基、2，2'-联氮-二（3-乙基-苯并噻唑-6-磺酸）二铵盐（2，2'-azinobis-3-ethylbenzthiazoline-6-sulphonate，ABTS⁺）和1，1-二苯基-2-三硝基苯肼（1，1-diphenyl-2-picrylhydrazyl，DPPH）的IC₅₀值分别为7.66和6.77 mg/mL、6.16和5.30 mg/mL、0.63和0.85 mg/mL、0.39和2.62 mg/mL。以RAW264.7细胞建立H₂O₂诱导损伤模型，细胞抗氧化结果表明：HFCP1和HFCP1-1处理组与模型组相比均显著增强细胞活力（P<0.05），浓度为100和200 mg/L时MDA含量与模型组相比均显著下降（P<0.05）；HFCP1浓度为100和200 mg/L时与模型组相比显著增加SOD和GSH-Px的活性（P<0.05）；HFCP1-1浓度为200 mg/L时与模型组相比显著增加SOD活性（P<0.05）；也能增加GSH-Px的活性，但未达到显著水平（P>0.05）。

Abstract:

To analyze the antioxidant activity of Hericium caput-medusae(Bull.:Fr.) Pers (HFC), an industrial extraction of H. erinaceus polysaccharide fermentation extract was taken as study object. Using dialysis method and DEAE sepharose fast flow column purification crude polysaccharide components HFCP1 and neutral polysaccharides HFCP1-1 were obtained. The composition and structure of monosaccharides were preliminarily analyzed by PMP pre-column derivatization-high performance liquid chromatography and infrared spectroscopy. The monosaccharides were mainly composed of glucose and galactose. The results showed that the IC₅₀ values of HFCP1 and HFCP1-1 to scavenge hydroxyl radical scavenging, superoxide anion scavenging, ABTS⁺ and DPPH were 7.66 and 6.77, 6.16 and 5.30, 0.63 and 0.85, 0.39 and 2.62 mg/mL, respectively. A H₂O₂-induced injury model was established in RAW264.7 cells, the results of anti-oxidation of cells showed that the cell viability of HFCP1 and HFCP1-1 treatment group were significantly increased compared with those the model group (P<0.05). The content of MDA was significantly decreased compared with the model group when it concentration was 100 and 200 mg/L (P<0.05). The activities of SOD and GSH-Px were significantly increased compared with those of the model group when HFCP1 concentrations was 100 and 200 mg/L (P<0.05). In conclusion, compared with model group, 200 mg/L HFCP1-1 significantly increased SOD activity (P<0.05), but also increased GSH-Px activity, but did not reach significant level (P>0.05).