灵芝子实体多糖提取工艺优化及对哮喘小鼠改善作用

Optimization of extraction process of polysaccharides from Ganoderma lucidum fruit body and improving effect on asthmatic mice

为优化灵芝子实体多糖提取工艺并初步研究其对哮喘小鼠的改善作用,以灵芝子实体多糖提取率为响应值,在单因素试验基础上,采用响应面法优化多糖提取工艺条件。通过卵清蛋白诱导哮喘小鼠模型,以地塞米松(5 mg/kg)为阳性对照,考察低、中、高剂量灵芝多糖(50 mg/kg、100 mg/kg、200 mg/kg)对哮喘小鼠肺组织病理形态、肺泡灌洗液白细胞及细胞分类计数以及白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平的影响。结果表明,灵芝多糖最优提取工艺为:超声功率400 W,超声时间40 min,料液比1∶15(g∶mL)。在此最佳工艺条件下,多糖提取率为(3.22±0.04)%。与模型组相比,高剂量灵芝多糖组可显著改善哮喘小鼠肺组织血管和支气管壁明显增厚、大量炎性细胞浸润等病理变化,肺泡灌洗液中白细胞总数、淋巴细胞、中性粒细胞、嗜酸性粒细胞数量以及IL-6、TNF-α的水平分别降低至(34.96±5.81)×104 CFU/mL、(25.61±3.59)×104 CFU/mL、(4.67±1.59)×104 CFU/mL、(3.06±1.131)×104 CFU/mL、(82.62±14.07)pg/mL、(208.16±19.60)ng/L。结果显示灵芝多糖提取物可改善哮喘小鼠的炎症反应。

To optimize the extraction process of polysaccharides from Ganoderma lucidum fruit body and preliminarily study its effect on asthmatic mice,using the extraction rate of polysaccharides from G.lucidum fruit body as response value,on the base of single-factor experiments,the polysaccharides extraction process was optimized by response surface methodology.Asthmatic mice model was induced by ovalbumin.Using dexamethasone(5 mg/kg)as the positive control,the effects of low-,medium-and high-dose polysaccharides from G.lucidum fruit body(50 mg/kg,100 mg/kg,200 mg/kg)on the pathomorphology of lung tissue,white blood cells and cell classification count,and the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in bronchoalveolar lavage fluid were investigated.The results showed that the optimal extraction process of polysaccharides from G.lucidum fruit body was ultrasound power 400 W,time 40 min,and material to liquid ratio 1∶15(g∶ml).Under these optimal conditions,the extraction rate of polysaccharides was(3.22±0.04)%.Compared with the model group,the pathological changes such as obvious thickening of blood vessels and bronchial walls in the lung tissue of asthmatic mice and massive infiltration of inflammatory cells were significantly improved in the high-dose G.lucidum polysaccharides group,and the numbers of white blood cells,lymphocytes,neutrophils,and eosinophils and the concentrations of IL-6 and TNF-αin bronchoalveolar lavage fluid were significantly reduced to(34.96±5.81)×104 CFU/ml,(25.61±3.59)×104 CFU/ml,(4.67±1.59)×104 CFU/ml,(3.06±1.131)×104 CFU/ml,(82.62±14.07)pg/ml and(208.16±19.60)ng/L,respectively.The results indicated that the polysaccharides from G.lucidum fruit body could improve the inflammatory response in asthmatic mice.