纳米三羟异黄酮Caco-2细胞吸收实验
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摘要
文章通过实验研究了牛血清白蛋白(bovine serum albumin,BSA)包裹大豆三羟异黄酮(genistein,Gen)纳米体在小肠吸收的可行性。构建Caco-2单细胞模拟小肠上皮细胞对Gen纳米体的吸收转移模型,利用苯酚红通透性试验以及碱性磷酸酶(alkaline phosphatase,AKP)活性检测验证模型,实验结果显示Gen纳米体在跨膜转运的过程中发生了代谢与转化,Gen纳米体在单层细胞模型的顶(apical,AP)侧与基底(basolateral,BL)侧的表观渗透系数Papp均高于Gen单体对照组,说明由BSA包裹的Gen纳米体比Gen单体较易被Caco-2细胞吸收。
Experimental study on the feasibility of absorption of bovine serum albumin(BSA)-encapsulated soybean genistein(Gen)nanoparticles by small intestine was carried out.Caco-2 single cells were used to simulate the uptake and translocation of Gen nanoparticles by small intestinal epithelial cells,and the phenol red permeation test and the alkaline phosphatase(AKP)activity detection were used to validate the model.The results showed that the Gen nanoparticles occurred with metabolism and transformation in the process of transmembrane transport,and the apparent permeability coefficients(Papp)of the Gen nanoparticles on the apical(AP)side and the basolateral(BL)side of the monolayer cell model were both higher than those of the Gen monomers.It is suggested that the BSA-encapsulated Gen nanoparticles are easier to be absorbed by Caco-2 cells than Gen monomers.
Experimental study on the feasibility of absorption of bovine serum albumin(BSA)-encapsulated soybean genistein(Gen)nanoparticles by small intestine was carried out.Caco-2 single cells were used to simulate the uptake and translocation of Gen nanoparticles by small intestinal epithelial cells,and the phenol red permeation test and the alkaline phosphatase(AKP)activity detection were used to validate the model.The results showed that the Gen nanoparticles occurred with metabolism and transformation in the process of transmembrane transport,and the apparent permeability coefficients(Papp)of the Gen nanoparticles on the apical(AP)side and the basolateral(BL)side of the monolayer cell model were both higher than those of the Gen monomers.It is suggested that the BSA-encapsulated Gen nanoparticles are easier to be absorbed by Caco-2 cells than Gen monomers.
引文
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[19]左克源.Caco-2细胞模型的建立及在口服药物吸收中的应用[D].武汉:华中科技大学2007.
[20] ARTURSSON P,KARLSSON J.Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial(Caco-2)cells[J].Biochemical&Biophysical Research Communi-cations,1991,175(3):880-885.
[21]孙海燕,廖晓慧,彭光华,等.Caco-2细胞模型及其在食品营养物质吸收研究中的新进展[J].时珍国医国药,2007,18(10):37-40.
[22]李红卫,王开萍,吴娱,等.猪肌肉组织中碱性磷酸酶的纯化及特性研究[J].合肥工业大学学报(自然科学版),2016,39(4):560-565.
[23]李晓丽.Caco-2细胞对不同形态锰的吸收及机制研究[D].北京:中国农业科学院,2013.
[24]王海玲.利用Caco-2细胞模型对黄酮类化合物吸收代谢的研究[D].长春:吉林大学,2006.
[25]王亚之,欧喜笑,郑颖.外排转运体和代谢酶与黄酮的相互作用及其对黄酮肠吸收影响的研究进展[J].中草药,2009,40(10):1659-1663.
[2] KIM J H,KIM Y J.Effects of genistein in combination with conjugated estrogens on endometrial hyperplasia and metabolic dysfunction in ovariectomized mice[J].Endocrine Journal,2015,62(6):531-542.
[3] ALNAKKASH L,JANJULIA T,PETERSON K,et al.Genistein and exercise do not improve cardiovascular risk factor in the ovariectomized rat[J].Climacteric the Journal of the International Menopause Society,2014,17(2):136-147.
[4]冯乙巳,潘宗琴,齐洪侠,等.一种黄豆甙元季胺盐衍生物合成及其活性研究[J].合肥工业大学学报(自然科学版),2010,33(12):1866-1869.
[5]谢玄.三羟基异黄酮壳聚糖纳米体的制备及其体内吸收、抗氧化研究[D].合肥:合肥工业大学,2012.
[6] WEISS J,TAKHISTOV P,MCCLEMENTS D J.Functional materials in food nanotechnology[J].Journal of Food Science,2006,71(9):R107-R116.
[7]刘世芸,徐超,冯乙巳,等.白蛋白聚乳酸缓释微球的制备及体外释放研究[J].合肥工业大学学报(自然科学版),2006,29(12):1584-1587.
[8]周瑞.牛血清白蛋白:槲皮素纳米颗粒的稳定性和细胞摄入及吸收的研究[D].北京:中国农业大学,2015.
[9]陈灏珠.实用内科学[M].14版.北京:人民卫生出版社,2013:1467-1478.
[10]MCCLUNG J A,KRUGER A L,FERRARIS A,et al.Usefulness of clopidogrel to protect against diabetes-induced vascular damage[J].American Journal of Cardiology,2010,105(7):1014-1018.
[11]夏亚飞,阎姝.仿制与进口氯吡格雷治疗急性冠脉综合征的成本效果分析[J].中国药房,2012,23(40):3801-3803.
[12] LI H,LI J,LIU L,et al.Elucidation of the intestinal absorption mechanism of celastrol using the Caco-2cell transwell model[J].Planta Medica,2016,82(13):1202-1207.
[13] GOULART A J,BASSAN J C,BARBOSA O A,et al.Transport of amino acids from milk whey by Caco-2cell monolayer after hydrolytic action of gastrointestinal enzymes[J].Food Research International,2014,63:62-70.
[14] RANALDI G,CONSALVO R,SAMBUY Y,et al.Permeability characteristics of parental and clonal human intestinal Caco-2cell lines differentiated in serum-supplemented and serum-free media[J].Toxicology in Vitro,2003,17(6):761-767.
[15]祝欣刚.色胺酮在Caco-2细胞模型上的吸收转运研究及Caco-2模型短期培养法的建立[D].上海:复旦大学,2010.
[16]康天舒.不同粒径纳米氧化锌对结肠腺癌Caco-2细胞的毒性作用研究[D].杭州:中国计量学院2014.
[17]王俊,徐倩倩,黄超,等.丝素蛋白/改性Fe3O4纳米粒子复合材料的制备[J].合肥工业大学学报(自然科学版),2012,35(10):1320-1323.
[18]邓纯博,刘学勇,刘冬妍.高浓度氧对肠上皮细胞Caco-2生长的影响[J].中国现代医学杂志,2009,19(13):1921-1923.
[19]左克源.Caco-2细胞模型的建立及在口服药物吸收中的应用[D].武汉:华中科技大学2007.
[20] ARTURSSON P,KARLSSON J.Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial(Caco-2)cells[J].Biochemical&Biophysical Research Communi-cations,1991,175(3):880-885.
[21]孙海燕,廖晓慧,彭光华,等.Caco-2细胞模型及其在食品营养物质吸收研究中的新进展[J].时珍国医国药,2007,18(10):37-40.
[22]李红卫,王开萍,吴娱,等.猪肌肉组织中碱性磷酸酶的纯化及特性研究[J].合肥工业大学学报(自然科学版),2016,39(4):560-565.
[23]李晓丽.Caco-2细胞对不同形态锰的吸收及机制研究[D].北京:中国农业科学院,2013.
[24]王海玲.利用Caco-2细胞模型对黄酮类化合物吸收代谢的研究[D].长春:吉林大学,2006.
[25]王亚之,欧喜笑,郑颖.外排转运体和代谢酶与黄酮的相互作用及其对黄酮肠吸收影响的研究进展[J].中草药,2009,40(10):1659-1663.