石斛多糖提取和对体外黑色素抑制、抗干燥损伤的影响

Extraction of Dendrobium Polysaccharides and Its Effects on Inhibition of Melanin and Resistance to Drying Damage in Vitro

用正交试验法优化铁皮石斛多糖的提取工艺条件,并探究了石斛多糖对B16细胞黑色素生成和HaCaT细胞内干燥损伤的影响。通过单因素实验考察了料液比、提取时间、提取次数和提取温度对多糖提取率的影响,进而进行正交试验,优化铁皮石斛多糖提取工艺条件;通过B16细胞实验,测定石斛多糖对细胞增殖率、细胞内酪氨酸酶的活性以及细胞内黑色素含量的影响;通过建立HaCaT细胞干燥损伤模型,测定多糖对细胞干燥损伤的影响。各因素对铁皮石斛多糖提取率影响的顺序为:提取次数>料液比>提取温度>提取时间,最佳提取条件是提取时间为60 min、料液比为1∶20、提取次数2次和提取温度为60℃,此工艺下石斛多糖的提取率为12.45%;石斛多糖(10,62.5,125,250,500μg/mL)对B16细胞活力无影响,在多糖质量浓度为500μg/mL时,酪氨酸酶活性为69.30%,黑色素含量为66.20%。石斛多糖(10,62.5,125,250,500μg/mL)对HaCaT细胞活力无影响,在多糖质量浓度为125μg/mL时,干燥损伤细胞的存活率提高了15.51%,达到了86.58%。本文优选建立了铁皮石斛多糖的最佳提取工艺,并发现石斛多糖可通过抑制细胞内酪氨酸酶活性来减少黑色素含量,建立HaCaT细胞干燥损伤模型,石斛多糖可以提高细胞在干燥损伤中的存活率,减少细胞干燥损伤造成的死亡。

In this study,the polysaccharide was extracted by hot water extraction from Dendrobium officinale.The polysaccharide extraction rate was used as an indicator.The ratio of extraction material to liquid,the extraction temperature,extraction times and the extraction time were the influencing factors.A single factor experiment was designed.A better extraction process was obtained through orthogonal experiments.The extraction rate of polysaccharides was 12.45%,and the optimal extraction conditions were:extraction time is 60 min,material-liquid ratio is 1:20,extraction times are 2 times,and extraction temperature is 60℃.The effects of polysaccharides on cell viability,tyrosinase activity and melanin content were determined by B16 cells experiment.By establishing a dry injury model in HaCaT cells,HaCaT cells dry injury model was established,and the cell viability of polysaccharides on cells in dry injury was determined.Different concentrations of polysaccharides(10,62.5,125,250,500μg/mL)can inhibit melanin production,in the polysaccharide concentration of 500μg/mL,the tyrosinase activity was 69.30%and the melanin content was 66.20%.Different concentrations of polysaccharides(10,62.5,125,250,500μg/mL)can reduce cell desiccation damage,and in the polysaccharide concentration of 125μg/mL,the survival rate of cell desiccation damage increases by 15.51%to 86.58%.Dendrobium officinale polysaccharide can reduce the melanin content by inhibiting tyrosinase activity,and can increase the survival rate of cells in desiccation damage and reduce the death of cells caused by desiccation damage.With the establishment of the HaCaT cells desiccation damage model,polysaccharides can improve the survival rate of cells in desiccation damage and reduce the death of cells caused by desiccation damage.