黑木耳分级多糖抗氧化活性及其相关生理功能研究
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摘要
黑木耳(Auricularia auricula)是著名的药食兼用真菌,其主要的生理活性成分是黑木耳多糖,具有抗肿瘤、调节免疫、抗疲劳、降血糖、降血脂、抗血栓等生理功能。
本文研究了黑木耳分级多糖的制备条件,并对黑木耳分级多糖进行了体外抗氧化,以及体内抗辐射、抗氧化、抗疲劳等生理功能研究,得到的主要结果如下：
1.通过单因素试验以及响应面分析,得到水溶性黑木耳多糖的最佳提取工艺条件为：最佳条件液料比为102：1、提取时间为2.1h、提取温度为88℃,多糖得率可达到15.1%。
2.用磷酸盐缓冲溶液制备了酸溶性黑木耳分级多糖和碱溶性黑木耳分级多糖。通过单因素试验和正交试验,确定了纯化水溶性黑木耳多糖的最佳洗脱条件为：径长比1：30、洗脱剂浓度0.5 mol/L、洗脱流速0.7mL/min。通过排阻层析,制备了水溶性黑木耳多糖纯品AAPⅠ-1。
3.通过对酸溶性黑木耳分级多糖APG6、水溶性黑木耳多糖AAPⅠ-1和碱溶性黑木耳分级多糖APG8对ABTS自由基、DPPH自由基、超氧自由基、羟自由基、过氧化氢自由基、还原力,对金属离子螯合和脂质过氧化抑制的研究,结果表明：APG8具有明显更强的作用,APG6次之,AAPⅠ-1比APG6稍弱,且相对分子量小的黑木耳分级多糖具有更强的抗氧化作用。
4.通过对酸溶性黑木耳分级多糖APG6-3、水溶性黑木耳多糖AAPⅠ-1和碱溶性黑木耳分级多糖APG8-3体内抗氧化作用的研究表明：APG8-3的体内抗氧化作用最强,APG6-3的作用稍弱,AAPⅠ-1的作用相对较弱,且高剂量组的抗氧化效果明显好于低剂量组。
5.通过对酸溶性黑木耳分级多糖APG6-3.水溶性黑木耳多糖AAPⅠ-1和碱溶性黑木耳分级多糖APG8-3抗辐射作用的研究表明：辐射引起的氧化损伤是辐射损伤的主要原因；各剂量组多糖对辐射引起的免疫功能下降和氧化损伤具有不同程度的保护作用;3组多糖在抗辐射作用方面各有其优势,且高剂量组的作用明显好于低剂量组。
6.通过对酸溶性黑木耳分级多糖APG6-3.水溶性黑木耳多糖AAPⅠ-1和碱溶性黑木耳分级多糖APG8-3抗疲劳作用的研究表明：疲劳的产生是能源耗竭、疲劳物质堵塞和氧自由基及其引起的脂质过氧化反应增强综合作用的结果；APG8-3对延长小鼠力竭游泳时间的作用最强,APG6-3次之,AAPⅠ-1相对较弱；APG8-3对提高和恢复小鼠肝糖原和肌糖原含量的作用最强,AAPⅠ-1次之,APG6-3相对较弱；APG8-3对降低小鼠BLA含量和BUN含量的作用最强,AAPⅠ-1和APG6-3的作用相近。
综上所述,碱溶性黑木耳分级多糖APG8-3在体外抗氧化以及体内抗辐射、抗氧化、抗疲劳等生理功能方面具有更强的作用,水溶性黑木耳多糖AAPⅠ-1和酸溶性黑木耳分级多糖APG6-3的作用基本相当,也都具有较强的生理功能,并且黑木耳分级多糖的抗辐射和抗疲劳功能主要是通过发挥其抗氧化作用来实现的。本文的研究结果为黑木耳多糖在医药、功能保健品、化妆品等领域的应用奠定了基础。
Auricularia auricular is well known as a edible fungus which has highly nutritious and officinal function. Polysaccharides of Auricularia auricular(AAP) are main function components of Auricularia auricular and have various functions such as antitumor, modulating immunity, antifatigue, decreasing blood sugar and blood fat, antithrombus and so on.
Preparation of fraction AAP and several functions including antioxidation in vitro and in vivo, anti irradiation and antifatigue, were studied in this paer. The main results of this paper as follows:
1. The optimal processing condition of water-soluble AAP, which ratio of liquid to solid 102:1, extraction time 2.1h、extraction temperature 88℃, was gained by single factor experiments and response surface analysis method. The yield of water-soluble AAP was 15.1%.
2. Acidity-soluble AAP and alkali-soluble AAP were prepared using phosphate buffer solution. The optimal elution conditions of purifying water-soluble AAP which were diameter-length ratio 1:30, eluent concentration 0.5mol/L, elution velocity 0.7mL/min, were obtained by single factor and orthogonal experiments.
3. The ability of acidity-soluble fraction AAP(APG6), water-soluble AAP (AAPⅠ-1) and alkali-soluble fraction AAP(APG8) scavenging ABTS radical, DPPH radical, O2- radical, OH radical, H2O2, reducing power and restraining metal ion chelation and lipid peroxidation were studied. The results showed that APG8 have higher effect, APG6 lower than APG8, AAP I-1 little lower than APG6. The components with lower relative molecular weight of APG6 and APG8 had better antioxidation function.
4. The antioxidation function in vivo of acidity-soluble AAP(APG6-3), AAP I-1 and alkali-soluble AAP(APG8-3) were studied. The results showed that APG8-3 have higher antioxidation function, APG6-3 little lower than APG8-3, AAPⅠ-1 lower than APG6-3. The antioxidation function of high dosage group obviously better than that of low dosage group. The results showed that fraction AAP with antioxidation in vitro had corresponding effect in vivo by correlation analysis between in vitro and in vivo.
5. The anti irradiation function of APG6-3, AAPⅠ-1 and APG8-3 were studied. The results showed that oxidative damage caused by irradiation was main damage of irradiation. Every dosage group all had protection effect about immunity decreasing and oxidation damage caused by irradiation. APG6-3, AAPⅠ-1 and APG8-3 had its own advantage respectively, and the anti irriadation function of high dosage group obviously better than that of low dosage group. The function of anti irradiation of AAP was accomplished by fulfilling its antioxidation function.
6. The antifatigue function of APG6-3, AAPⅠ-1 and APG8-3 were studied. The results showed that fatigue caused by energy depletion, fatigue production blocking and oxygen free radical and lipid peroxidation reaction enhancement. APG8-3 had better function of prolonging forced-swimming time of mice, APG6-3 lower than APG8-3, AAPⅠ-1 lower than APG6-3; APG8-3 had better function of increasing LG and MG contents, AAPⅠ-1 than APG8-3, APG6-3 lower than AAPⅠ-1; APG8-3 had better function of decreasing BLA and BUN contents, AAPⅠ-1 and APG6-3 lower than APG8-3.
To sum up, APG8-3 had better function than AAPⅠ-1 and APG6-3 about antioxidation in vitro and in vivo, anti irradiation and antifatigue, and AAPⅠ-1 and APG6-3 had approximate function. The function of anti irradiation and antifatigue of AAP was accomplished by fulfilling its antioxidation function. The results of this paper lay a foundation for AAP exploitation and application in medicine, health food, cosmetic and so on.
本文研究了黑木耳分级多糖的制备条件,并对黑木耳分级多糖进行了体外抗氧化,以及体内抗辐射、抗氧化、抗疲劳等生理功能研究,得到的主要结果如下：
1.通过单因素试验以及响应面分析,得到水溶性黑木耳多糖的最佳提取工艺条件为：最佳条件液料比为102：1、提取时间为2.1h、提取温度为88℃,多糖得率可达到15.1%。
2.用磷酸盐缓冲溶液制备了酸溶性黑木耳分级多糖和碱溶性黑木耳分级多糖。通过单因素试验和正交试验,确定了纯化水溶性黑木耳多糖的最佳洗脱条件为：径长比1：30、洗脱剂浓度0.5 mol/L、洗脱流速0.7mL/min。通过排阻层析,制备了水溶性黑木耳多糖纯品AAPⅠ-1。
3.通过对酸溶性黑木耳分级多糖APG6、水溶性黑木耳多糖AAPⅠ-1和碱溶性黑木耳分级多糖APG8对ABTS自由基、DPPH自由基、超氧自由基、羟自由基、过氧化氢自由基、还原力,对金属离子螯合和脂质过氧化抑制的研究,结果表明：APG8具有明显更强的作用,APG6次之,AAPⅠ-1比APG6稍弱,且相对分子量小的黑木耳分级多糖具有更强的抗氧化作用。
4.通过对酸溶性黑木耳分级多糖APG6-3、水溶性黑木耳多糖AAPⅠ-1和碱溶性黑木耳分级多糖APG8-3体内抗氧化作用的研究表明：APG8-3的体内抗氧化作用最强,APG6-3的作用稍弱,AAPⅠ-1的作用相对较弱,且高剂量组的抗氧化效果明显好于低剂量组。
5.通过对酸溶性黑木耳分级多糖APG6-3.水溶性黑木耳多糖AAPⅠ-1和碱溶性黑木耳分级多糖APG8-3抗辐射作用的研究表明：辐射引起的氧化损伤是辐射损伤的主要原因；各剂量组多糖对辐射引起的免疫功能下降和氧化损伤具有不同程度的保护作用;3组多糖在抗辐射作用方面各有其优势,且高剂量组的作用明显好于低剂量组。
6.通过对酸溶性黑木耳分级多糖APG6-3.水溶性黑木耳多糖AAPⅠ-1和碱溶性黑木耳分级多糖APG8-3抗疲劳作用的研究表明：疲劳的产生是能源耗竭、疲劳物质堵塞和氧自由基及其引起的脂质过氧化反应增强综合作用的结果；APG8-3对延长小鼠力竭游泳时间的作用最强,APG6-3次之,AAPⅠ-1相对较弱；APG8-3对提高和恢复小鼠肝糖原和肌糖原含量的作用最强,AAPⅠ-1次之,APG6-3相对较弱；APG8-3对降低小鼠BLA含量和BUN含量的作用最强,AAPⅠ-1和APG6-3的作用相近。
综上所述,碱溶性黑木耳分级多糖APG8-3在体外抗氧化以及体内抗辐射、抗氧化、抗疲劳等生理功能方面具有更强的作用,水溶性黑木耳多糖AAPⅠ-1和酸溶性黑木耳分级多糖APG6-3的作用基本相当,也都具有较强的生理功能,并且黑木耳分级多糖的抗辐射和抗疲劳功能主要是通过发挥其抗氧化作用来实现的。本文的研究结果为黑木耳多糖在医药、功能保健品、化妆品等领域的应用奠定了基础。
Auricularia auricular is well known as a edible fungus which has highly nutritious and officinal function. Polysaccharides of Auricularia auricular(AAP) are main function components of Auricularia auricular and have various functions such as antitumor, modulating immunity, antifatigue, decreasing blood sugar and blood fat, antithrombus and so on.
Preparation of fraction AAP and several functions including antioxidation in vitro and in vivo, anti irradiation and antifatigue, were studied in this paer. The main results of this paper as follows:
1. The optimal processing condition of water-soluble AAP, which ratio of liquid to solid 102:1, extraction time 2.1h、extraction temperature 88℃, was gained by single factor experiments and response surface analysis method. The yield of water-soluble AAP was 15.1%.
2. Acidity-soluble AAP and alkali-soluble AAP were prepared using phosphate buffer solution. The optimal elution conditions of purifying water-soluble AAP which were diameter-length ratio 1:30, eluent concentration 0.5mol/L, elution velocity 0.7mL/min, were obtained by single factor and orthogonal experiments.
3. The ability of acidity-soluble fraction AAP(APG6), water-soluble AAP (AAPⅠ-1) and alkali-soluble fraction AAP(APG8) scavenging ABTS radical, DPPH radical, O2- radical, OH radical, H2O2, reducing power and restraining metal ion chelation and lipid peroxidation were studied. The results showed that APG8 have higher effect, APG6 lower than APG8, AAP I-1 little lower than APG6. The components with lower relative molecular weight of APG6 and APG8 had better antioxidation function.
4. The antioxidation function in vivo of acidity-soluble AAP(APG6-3), AAP I-1 and alkali-soluble AAP(APG8-3) were studied. The results showed that APG8-3 have higher antioxidation function, APG6-3 little lower than APG8-3, AAPⅠ-1 lower than APG6-3. The antioxidation function of high dosage group obviously better than that of low dosage group. The results showed that fraction AAP with antioxidation in vitro had corresponding effect in vivo by correlation analysis between in vitro and in vivo.
5. The anti irradiation function of APG6-3, AAPⅠ-1 and APG8-3 were studied. The results showed that oxidative damage caused by irradiation was main damage of irradiation. Every dosage group all had protection effect about immunity decreasing and oxidation damage caused by irradiation. APG6-3, AAPⅠ-1 and APG8-3 had its own advantage respectively, and the anti irriadation function of high dosage group obviously better than that of low dosage group. The function of anti irradiation of AAP was accomplished by fulfilling its antioxidation function.
6. The antifatigue function of APG6-3, AAPⅠ-1 and APG8-3 were studied. The results showed that fatigue caused by energy depletion, fatigue production blocking and oxygen free radical and lipid peroxidation reaction enhancement. APG8-3 had better function of prolonging forced-swimming time of mice, APG6-3 lower than APG8-3, AAPⅠ-1 lower than APG6-3; APG8-3 had better function of increasing LG and MG contents, AAPⅠ-1 than APG8-3, APG6-3 lower than AAPⅠ-1; APG8-3 had better function of decreasing BLA and BUN contents, AAPⅠ-1 and APG6-3 lower than APG8-3.
To sum up, APG8-3 had better function than AAPⅠ-1 and APG6-3 about antioxidation in vitro and in vivo, anti irradiation and antifatigue, and AAPⅠ-1 and APG6-3 had approximate function. The function of anti irradiation and antifatigue of AAP was accomplished by fulfilling its antioxidation function. The results of this paper lay a foundation for AAP exploitation and application in medicine, health food, cosmetic and so on.
引文
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