负载槲皮素的酶法糖基化酪蛋白复合纳米粒子的性能研究

刘尧; 樊永康; 吴晓琴; 沈建福

doi:10.13386/j.issn1002-0306.2020110211

负载槲皮素的酶法糖基化酪蛋白复合纳米粒子的性能研究

Study on Properties of Quercetin-loaded Enzymatic Glycosylated Casein Composite Nanoparticles

摘要

摘要: 制备负载槲皮素的酪蛋白纳米粒子（_n-Cas-Que）和转谷氨酰胺酶法糖基化酪蛋白纳米粒子（_n-Cas-Cos-Que），探究其贮藏稳定性、体外抗消化性、生物可及度、抗氧化性及对人前列腺癌PC-3细胞的生长抑制效果。结果表明：在4 ℃避光、25 ℃避光、25 ℃自然光照条件下分别贮藏60 d，两种纳米粒子中槲皮素的保留率分别为80.78%、91.30%；66.87%、79.25%；22.18%、34.33%，即_n-Cas-Cos-Que纳米粒子能够更好的保护槲皮素。体外模拟消化结果显示，_n-Cas-Cos-Que比_n-Cas-Que具有更好的抗消化性和更高的生物可及度。在体外模拟消化180 min后，槲皮素水溶液、_n-Cas-Que和_n-Cas-Cos-Que的生物可及度分别为10.13%、27.65%和48.91%。采用DPPH法和ABTS法探究两种纳米粒子的抗氧化活性，_n-Cas-Cos-Que、_n-Cas-Que对DPPH和ABTS的清除率分别为58.71%、53.82%和56.98%、48.34%，即_n-Cas-Cos-Que比_n-Cas-Que具有更好的抗氧化性。体外抑制PC-3细胞试验的结果表明，随着槲皮素浓度的增加，Que、_n-Cas-Que、_n-Cas-Cos-Que对PC-3细胞的抑制效果逐渐增强。当槲皮素的浓度达到80 µg/mL时，用Que、_n-Cas-Que、_n-Cas-Cos-Que体系处理的PC-3细胞的存活率分别为73.25%、56.84%、49.88%，且三种体系处理后的细胞存活率有显著性差异（P<0.05），说明与Que和_n-Cas-Que相比，_n-Cas-Cos-Que纳米粒子对PC-3细胞有更好的抑制效果。本实验结果表明了酪蛋白纳米粒子能够较好的保护槲皮素，且酶法糖基化能提高其对槲皮素的保护效果，并提高其抗消化性、生物可及度、抗氧化性和抗癌活性。

Abstract: Objective: To prepare quercetin loaded casein nanoparticles (_n-Cas-Que) and transglutaminase glycosylated casein nanoparticles (_n-Cas-Cos-Que), and explore their storage stability, in vitro anti digestibilitiy, bioaccessibility, anti-oxidation and inhibitory effect on human prostate cancer PC-3 cells. The results showed that when stored in 4 ℃ avoid light, 25 ℃ avoid light and 25 ℃ natural light for 60 days, the retention rates of quercetin in _n-Cas-Que and _n-Cas-Cos-Que were 80.78%, 91.30%, 66.87%, 79.25%, 22.18%, 34.33%, respectively, which indicated that _n-Cas-Cos-Que nanoparticles could protect quercetin better.The results of anti-digestion pointed out that _n-Cas-Cos-Que had better anti-digestibility and higher bioavailability than _n-Cas-Que. After 180 min of simulated digestion in vitro, the bioaccessibility of quercetin aqueous solution, _n-Cas-Que and _n-Cas-Cos-Que were 10.13%, 27.65% and 48.91%, respectively. DPPH method and ABTS method were used to investigate the antioxidant activity of the two nanoparticles. The scavenging rates of _n-Cas-Cos-Que and _n-Cas-Que on DPPH and ABTS were 58.71%, 53.82% and 56.98% and 48.34%, respectively, which indicated that _n-Cas-Cos-Que had better antioxidant activity than _n-Cas-Que. The inhibition of human prostate cancer PC-3 cells expressed that with the increase of quercetin concentration, the inhibitory effects of Que, _n-Cas-Que and _n-Cas-Cos-Que on PC-3 cells were gradually enhanced. When the concentration of quercetin reached 80 μg/mL, the survival rates of PC-3 cells treated with Que, _n-Cas-Que and _n-Cas-Cos-Que were 73.25%, 56.84% and 49.88% respectively, and there were significant differences among these survival rates (P<0.05). Compared with Que and _n-Cas-Que, _n-Cas-Cos-Que nanoparticles had better inhibitory effect on PC-3 cells. The results showed that casein nanoparticles can protect quercetin well, and enzymatic glycosylation can improve its protective effect on quercetin, and raise its anti digestion and biological accessibility, as well as antioxidant and anti-cancer activity.