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使用离子交换和凝胶过滤色谱法从富硒螺旋藻中提纯含硒藻蓝蛋白。通过含硒藻蓝蛋白与藻蓝蛋白对4种不同的自由基清除能力的比较,即总抗氧化能力、清除DPPH自由基、过氧化物阴离子和引发红细胞溶血的过氧化氢自由基能力测定,从而对含硒藻蓝蛋白的抗氧化活性进行评价。结果显示,经提纯后的含硒藻蓝蛋白在某一特定剂量下与藻蓝蛋白在清除ABTS、DPPH、过氧化物阴离子和AAPH自由基能力相比,都表现出更好的抗氧化作用。评价提纯后的含硒藻蓝蛋白和藻蓝蛋白在抑制人体恶性黑素瘤细胞(A-375)的生长和增殖的作用。结果表明,提纯后的含硒藻蓝蛋白和藻蓝蛋白的IC50分别在44.5μM和65.9μM能显著地抑制A-375细胞的增殖。使用流体血细胞计数分析和末端脱氧核苷酸转移酶和脱氧尿三磷酸缺口末端标记对A-375细胞凋亡的诱发进行测定。
Selenium - depleted phycocyanin was purified from Se - enriched Spirulina by ion exchange and gel filtration chromatography. Through the comparison of four different free radical scavenging abilities, the total antioxidant capacity, DPPH radical scavenging, peroxide anion and the hydrogen peroxide radical activity of erythrocyte hemolysis, Thus, the antioxidant activity of the phycocyanin-containing selenium was evaluated. The results showed that purified phycocyanin showed a better antioxidant activity than phycocyanin at ABTS, DPPH, peroxide anion and AAPH radical scavenging ability at a certain dosage. The purified phytoncocyanin and phycocyanin were evaluated for their ability to inhibit the growth and proliferation of human malignant melanoma cells (A-375). The results showed that the IC50 of purified phycocyanin and phycocyanin at 44.5 μM and 65.9 μM, respectively, significantly inhibited the proliferation of A-375 cells. Apoptosis induction in A-375 cells was assayed by flow cytometry and terminal deoxynucleotidyl transferase and deoxyurone triphosphate nick end labeling.