铆钉菇黑色素抗氧化活性及对秀丽隐杆线虫抗衰老与应激损伤的作用

Antioxidant Activities of Melanin from Gomphidius viscidus and Its Anti-aging, Anti-stress Injury Effects on Caenorhabditis elegans

研究了铆钉菇D1胞外黑色素分子结构、体内外抗氧化、抗衰老及应激损伤作用。通过紫外-可见光谱、红外光谱及扫描电镜对铆钉菇黑色素结构特性进行分析;通过测定羟基、1,1-二苯基-2-三硝基苯肼(DPPH)、超氧阴离子自由基清除能力及铁氰化钾还原力评价体外抗氧化能力;通过秀丽隐杆线虫自然衰老、过氧化氢、高温及紫外线等应激损伤模型,评价黑色素抗衰老及应激损伤的作用;通过测定秀丽隐杆线虫体内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及丙二醛(MDA)含量评价其体内抗氧化能力。结果表明:铆钉菇黑色素为无定型结构,具有典型的黑色素紫外-可见光及红外吸收光谱特征;羟基、DPPH及超氧阴离子自由基清除率最高达到68.42%,67.1%,34.3%;1.00 g/L的铆钉菇黑色素可显著提高线虫的自然寿命(P<0.05),0.50~1.00 g/L的铆钉菇黑色素可显著提高过氧化氢应激下线虫的平均寿命(P<0.01),0.50~4.00 g/L的铆钉菇黑色素可显著提高热应激下线虫的平均寿命(P<0.01),0.25~4.00 g/L的铆钉菇黑色素可显著提高紫外线应激下线虫的平均寿命(P<0.01);0.50~1.00 g/L的铆钉菇黑色素,可显著提高线虫体内SOD(P<0.05)和CAT活力(P<0.01),降低MDA含量(P<0.01)。结论:铆钉菇胞外黑色素具有典型的黑色素分子结构,良好的体内外抗氧化能力及抗衰老与应激损伤作用。

The structure characteristics,antioxidant activity and anti-aging of melanin from Gomphidius viscidus D1 were studied.Ultraviolet-visible spectroscopy,infrared spectroscopy and scanning electron microscope(SEM)were used to evaluate structure characteristics;Hydroxyl,1,1-diphenyl-2-picrylhydrazyl(DPPH),superoxide anion radical scavenging assays,and potassium ferricyanide reducing power were used to evaluate antioxidant activity in vitro;Natural aging,hydrogen peroxide,heat and ultraviolet damage models were used to evaluate anti-aging and anti-stress injuries on Caenorhabditis elegans;SOD,CAT activities and MDA content were measured to evaluate antioxidant activity in vivo.The results showed that melanin from Gomphidius viscidus is an amorphous structure,it has typical ultraviolet visible and infrared absorption spectra of melanin;It has good scavenging activities,the DPPH,hydroxyl radical and superoxide anion scavenging rates reached 67.1%,68.42%and 34.3%;The natural lifespan of Caenorhabditis elegans was prolonged significantly by 1.00 g/L melanin(P<0.05),lifespan under oxidative stress,heat stress and ultraviolet radiation stress could be prolonged by melanin at the mass concentration of 0.50-1.00 g/L,0.50-4.00 g/L and 0.25-4.00 g/L respectively(P<0.01);At 0.50-1.00 g/L,the SOD(P<0.05),CAT(P<0.01)activities were significantly increased,and MDA content(P<0.01)was significantly decreased.In conclusion,melanin from Gomphidius viscidus has good antioxidant activity both in vitro and in vivo and has good anti-aging and anti-stress injury effects.