以香港牡蛎蛋白为原料,采用中性蛋白酶进行酶解制备牡蛎蛋白酶解产物(oyster protein enzymatic hydrolysate,OPEH),逐级通过8 k、5 k、3 kDa分子筛超滤膜,共截留出<3 kDa(OPEH-1)、3 k ~5 kDa(OPEH-2)、5 k~8 kDa(OPEH-3)和>8 kDa(OPEH-4)等超滤组分。首先对比OPEH及4个超滤组分的体外总抗氧化能力,然后在中波紫外线(ultraviolet B,UVB)诱导的HaCaT(人永生化角层细胞系)光老化细胞模型上,评价其抗皮肤光老化活性,对可能机制进行初步探索。体外总抗氧化能力结果显示,OPEH-1的ABTS阳离子自由基半清除质量浓度(IC50)最低为156.083 μg/mL,具有最佳的体外抗氧化能力。细胞实验结果显示,与OPEH以及其他超滤组分相比,OPEH-1在质量浓度>50 μg/mL 后就能显著地提升UVB辐照后HaCaT细胞的活力,能逆转UVB辐射后的HaCaT细胞形态改变,抑制细胞内活性氧的产生,促进I型胶原蛋白前体的分泌。研究发现,OPEH-1在UVB辐射诱导的HaCaT光老化细胞模型上,具有最好的抗光老化活性,这种活性可能与OPEH-1具有较好的抗氧化能力以及促进I胶原蛋白前体的表达相关。
Hongkong oyster protein was hydrolyzed by neutral protease firstly. Subsequently, enzymatic oyster protein hydrolysate (OPEH) was gradually ultra-filtrated by 8, 5 and 3 kDa membrane with molecular weight cutoffs to separate four fractions, including<3 kDa (OPEH-1), 3-5 kDa (OPEH-2), 5-8 kDa (OPEH-3), and >8kDa (OPEH-3) fractions. Finally, the total antioxidant capacity of OPEH and four fractions were measured in vitro, and the anti-skin photoaging activities were tested in an UVB-induced HaCaT photoaged cell model. The results of total antioxidant capacity showed that OPEH-1 had the lowest ABTS cation free radical semi-scavenging concentration (IC50), exerted the best antioxidant effect in vitro. The results from cellular experiment indicated that compared with OPEH and other fractions. OPEH-1 significantly improved cell viability starting the concentration in 50 μg/mL, reversed morphological changes of UVB-irradiated HaCaT cells, inhibited the production of intracellular reactive oxygen ROS, while increased the production of extracellular pro-collagen I. The anti-skin photoaging effect of OPEH-1 was possibly associated with its anti-antioxidant property and the activity of improving extracellular pro-collagen I expression.
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