姬松茸多糖的提取及其对D-半乳糖诱导衰老小鼠的免疫调节作用
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摘要
建立姬松茸多糖(Agaricus blazei polysaccharides,ABP)的提取方法,并研究姬松茸多糖对D-半乳糖诱导的衰老小鼠免疫调节作用。采用水提醇沉法提取姬松茸多糖,通过单因素实验和正交试验筛选并优化姬松茸多糖的最佳提取工艺;采用D-半乳糖(400 mg/kg)构建衰老小鼠模型,将小鼠随机分为5组,空白对照组(CON)、模型组(MOD)、姬松茸多糖低剂量组(ABP-L,200 mg/kg/d)、姬松茸多糖中剂量组(ABP-M,400 mg/kg/d)、姬松茸多糖高剂量组(ABP-H,800 mg/kg/d);分别测定各组小鼠胸腺和脾脏指数;CCK-8法测定ConA诱导的小鼠脾淋巴细胞增殖转化;ELISA法检测血清中TNF-α、IL-6含量。结果表明,姬松茸多糖最佳提取工艺为:提取温度100℃,提取时间3 h,料液比1∶20 g/mL,提取次数3次,此时姬松茸多糖的平均得率为8.814%±0.109%;与模型组相比,姬松茸多糖高剂量组显著提高小鼠脾脏(p<0.01)和胸腺指数(p<0.05),姬松茸多糖高剂量组极显著促进衰老小鼠脾细胞增殖(p<0.01),增加衰老小鼠血清中TNF-α(p<0.01)、IL-6(p<0.01)含量。姬松茸多糖对D-半乳糖诱导的衰老小鼠模型具有免疫增强作用。
To establish the extraction method of Agaricus blazei polysaccharides(ABP),and study the immunomodulatory effects of Agaricus blazei polysaccharides on D-galactose-induced aging mice.Agaricus blazei polysaccharides were extracted by water extraction and alcohol precipitation. The extraction process of Agaricus blazei polysaccharides was screened and optimized by single factor experiments and orthogonal experiment. The aging mouse model was constructed by D-galactose(400 mg/kg).The mice were randomly divided into 5 groups:Blank control group(CON),model group(MOD),Agaricus blazei polysaccharides low dose group(ABP-L,200 mg/kg/d),Agaricus blazei polysaccharides medium dose group(ABP-M,400 mg/kg/d),high dose group of Agaricus blazei polysaccharides(ABP-H,800 mg/kg/d). After the experiment,the thymus and spleen index of each group were determined. The proliferation and transformation of mouse spleen lymphocytes induced by ConA were determined by CCK-8 method. The contents of TNF-α and IL-6 in serum were detected by ELISA. The results showed that the optimum extraction process of Agaricus blazei polysaccharides was as follows:Extraction temperature 100 ℃,extraction time 3 h,ratio of material to liquid 1∶20 g/mL,extraction times 3 times,the average yield of Agaricus blazei polysaccharides was 8.814%±0.109%.Compared with the model group,Agaricus blazei polysaccharides high dose group and ultrafine powder group significantly improved mice spleen(p<0.01)and thymus index(p<0.05),Agaricus blazei polysaccharides extremely significantly promoted spleen cell proliferation in aging mice(p<0.01),and increased TNF-α(p<0.01)and IL-6 in serum of aging mice(p<0.01).Agaricus blazei polysaccharides had immunomodulatory effect on D-galactose-induced aging mouse model.
To establish the extraction method of Agaricus blazei polysaccharides(ABP),and study the immunomodulatory effects of Agaricus blazei polysaccharides on D-galactose-induced aging mice.Agaricus blazei polysaccharides were extracted by water extraction and alcohol precipitation. The extraction process of Agaricus blazei polysaccharides was screened and optimized by single factor experiments and orthogonal experiment. The aging mouse model was constructed by D-galactose(400 mg/kg).The mice were randomly divided into 5 groups:Blank control group(CON),model group(MOD),Agaricus blazei polysaccharides low dose group(ABP-L,200 mg/kg/d),Agaricus blazei polysaccharides medium dose group(ABP-M,400 mg/kg/d),high dose group of Agaricus blazei polysaccharides(ABP-H,800 mg/kg/d). After the experiment,the thymus and spleen index of each group were determined. The proliferation and transformation of mouse spleen lymphocytes induced by ConA were determined by CCK-8 method. The contents of TNF-α and IL-6 in serum were detected by ELISA. The results showed that the optimum extraction process of Agaricus blazei polysaccharides was as follows:Extraction temperature 100 ℃,extraction time 3 h,ratio of material to liquid 1∶20 g/mL,extraction times 3 times,the average yield of Agaricus blazei polysaccharides was 8.814%±0.109%.Compared with the model group,Agaricus blazei polysaccharides high dose group and ultrafine powder group significantly improved mice spleen(p<0.01)and thymus index(p<0.05),Agaricus blazei polysaccharides extremely significantly promoted spleen cell proliferation in aging mice(p<0.01),and increased TNF-α(p<0.01)and IL-6 in serum of aging mice(p<0.01).Agaricus blazei polysaccharides had immunomodulatory effect on D-galactose-induced aging mouse model.
引文
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[2]张卉,刘长江.姬松茸(Agaricus blazei Murill)生理活性物质的研究进展[J].沈阳农业大学学报,2003(1):59-62.
[3]李晶,魏健.姬松茸多糖增强免疫功能及抗疲劳作用的研究[J].食品研究与开发,2017,38(5):201-204.
[4]张蓉娇,吴天祥.姬松茸多糖及其生物活性研究进展[J].贵州农业科学,2009,37(6):108-110.
[5]雷红,孙汉巨,姜绍通.菜籽饼粕多糖对D-半乳糖衰老小鼠的抗氧化及免疫调节作用[J].食品科学,2010,31(13):283-286.
[6]瞿俊勇,田梦,贺建华,等.白花蛇舌草多糖体外免疫活性研究[J].中兽医医药杂志,2015,34(3):17-20.
[7]张芮琪,陈正礼,罗启慧.黄芪多糖干预环磷酰胺所致免疫抑制小鼠的免疫功能[J].中国实验动物学报,2015,23(4):389-394.
[8]瞿俊勇.白花蛇舌草多糖提取工艺及免疫调节作用研究[D].长沙:湖南农业大学,2015.
[9]Ruonan B,Yaqin S,Shuzhen Z,et al.Simple nanoliposomes encapsulating Lycium barbarum polysaccharides as adjuvants improve humoral and cellular immunity in mice[J].International Journal of Nanomedicine,2017,173(12):6289-6301.
[10]周意,栾洁,蒋静娴,等.玛咖多糖对D-半乳糖衰老模型小鼠免疫器官的保护作用[J].中国实验方剂学杂志,2016,22(19):121-125.
[11]单颖,姜东,潘兴瑜,等.玉竹多糖对衰老模型鼠免疫功能的影响[J].中国老年学杂志,2007(1):20-22.
[12]赵波,张玉玲,咚继铭.赤苞果多糖对免疫抑制小鼠免疫功能的影响[J].时珍国医国药,2010,21(7):1627-1629.
[13]Jinzhu M A,Baifen S,Hongsheng R,et al.Effect of berberine hydrochloride on the proliferation,apoptosis and cytokine secretion of splenocytes in mice[J].Chinese Journal of Cellular and Molecular Immunology,2013,29(8):802-804.
[14]王思芦,汪开毓,陈德芳.食用真菌多糖免疫调节作用及其机制研究进展[J].动物医学进展,2012,33(11):104-108.
[15]王柳萍,沈茂杰,杨斌,等.牛大力多糖对小鼠脾细胞增殖及分泌细胞因子的影响[J].医药导报,2017,36(5):480-483.
[16]陈婉.蛹虫草抗氧化活性多糖制备技术研究[D].温州:温州大学,2016.
[17]王夏梅.姬松茸子实体多糖分离纯化、结构表征和免疫活性分析[D].无锡:江南大学,2016.
[18]周国海,张泳,赵力超,等.蛹虫草多糖提取纯化工艺研究[J].食品与机械,2014,30(5):220-224.
[19]彭勇胜.姬松茸多糖的制备及其抗氧化、保肝作用研究[D].武汉:华中农业大学,2011.
[20]Huang C,Song K,Ma W,et al.Immunomodulatory mechanism of bushen huoxue recipe alleviates cyclophosphamide-induced diminished ovarian reserve in mouse model[J].Journal of Ethnopharmacology,2017,208:44-56.