基于尿苷二磷酸葡萄糖醛酸转移酶1解毒通路探讨枸杞汁对邻苯二甲酸二(2-乙基己基)酯代谢的干预作用
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摘要
目的:通过大鼠毒性干预实验观察枸杞汁对邻苯二甲酸二(2-乙基己基)酯(di-(2-ethylhexyl)phthalate,DEHP)染毒大鼠肝脏中尿苷二磷酸葡萄糖醛酸转移酶1(uridine diphosphate glucuronosyltransferase 1,UGT1)活性及代谢解毒的影响,探讨枸杞汁对DEHP代谢的干预效果。方法:60只雄性大鼠随机分为对照组、DEHP组和枸杞汁干预组,每组20只,DEHP组和干预组在DEHP一次染毒(3 000 mg/kg mb)后连续7 d分别灌胃生理盐水和枸杞汁,对照组则给予同等体积的芝麻油或生理盐水,期间每天收集大鼠24 h尿液。在染毒1、3、5、7 d后各组分别随机处死5只。采用试剂盒检测肝脏UGT1的活力,高效液相色谱法检测血清中的邻苯二甲酸单(2-乙基己基)酯(mono-(2-ethylhexyl)phthalate,MEHP)以及尿液MEHP和邻苯二甲酸(phthalic acid,PA)含量。结果:枸杞汁干预组大鼠第5天UGT1活性显著高于DEHP组和对照组(P<0.05),且相对于DEHP组血清中MEHP含量减少而尿液中MEHP和PA含量增加。结论:枸杞汁可能通过提高UGT1活力促进了DEHP的代谢与排泄,发挥了解毒效应,将来可能作为一种预防或对抗邻苯二甲酸酯类物质或其他有毒化学物潜在危害的保健药材或功能食品用于人群的健康防护。
Purpose: This study aimed to explore the intervention effect of Lycium barbarum juice(LBJ) on the metabolism of di-(2-ethylhexyl) phthalate(DEHP) by analyzing the activity of uridine diphosphate glucuronosyltransferase 1(UGT1) and metabolic detoxification profile in the liver of rats exposed to DEHP. Methods: Sixty Sprague Dawley male rats were randomly divided into three groups(n = 20 each): control group, DEHP group and LBJ group. The DEHP and LBJ groups were administered with normal saline and LBJ respectively by gavage once daily for 7 consecutive days after receiving a single dose of 3 000 mg/kg mb DEHP dissolved in sesame oil. The control group was treated with normal saline for 7 days after receiving the same volume of sesame oil of. urine samples were collected for 24 hours daily during the experimental period. Totally 5 rats were sacrificed randomly in each group after 1, 3, 5 and 7 days of exposure. The activity of UGT1 was determined by an enzyme linked immunosorbent assay kit while the contents of mono-(2-ethylhexyl) phthalate(MEHP) in serum and urine and phthalic acid(PA) in urine were detected by high performance liquid chromatography. results: The activity of UGT1 in the LBJ group significantly increased on the 5 th day compared to the DEHP and control groups(P < 0.05). The content of serum MEHP in the LBJ group decreased significantly, whereas the contents of urine MEHP and PA increased compared to DEHP group. Conclusion: LBJ can promote the metabolism and excretion of DEHP through improving the activity of UGT1, making it a healthcare product or functional food against the potential hazard of phthalate esters(PAEs) or other poisonous chemicals.
Purpose: This study aimed to explore the intervention effect of Lycium barbarum juice(LBJ) on the metabolism of di-(2-ethylhexyl) phthalate(DEHP) by analyzing the activity of uridine diphosphate glucuronosyltransferase 1(UGT1) and metabolic detoxification profile in the liver of rats exposed to DEHP. Methods: Sixty Sprague Dawley male rats were randomly divided into three groups(n = 20 each): control group, DEHP group and LBJ group. The DEHP and LBJ groups were administered with normal saline and LBJ respectively by gavage once daily for 7 consecutive days after receiving a single dose of 3 000 mg/kg mb DEHP dissolved in sesame oil. The control group was treated with normal saline for 7 days after receiving the same volume of sesame oil of. urine samples were collected for 24 hours daily during the experimental period. Totally 5 rats were sacrificed randomly in each group after 1, 3, 5 and 7 days of exposure. The activity of UGT1 was determined by an enzyme linked immunosorbent assay kit while the contents of mono-(2-ethylhexyl) phthalate(MEHP) in serum and urine and phthalic acid(PA) in urine were detected by high performance liquid chromatography. results: The activity of UGT1 in the LBJ group significantly increased on the 5 th day compared to the DEHP and control groups(P < 0.05). The content of serum MEHP in the LBJ group decreased significantly, whereas the contents of urine MEHP and PA increased compared to DEHP group. Conclusion: LBJ can promote the metabolism and excretion of DEHP through improving the activity of UGT1, making it a healthcare product or functional food against the potential hazard of phthalate esters(PAEs) or other poisonous chemicals.
引文
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[11]王丽,袁晶,张荣.邻苯二甲酸二(2-乙基己基)酯环境暴露与人群健康研究进展[J].环境与健康杂志,2009,26(5):465-467.
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[16]柳春红,张明明,赖玉婷,等.邻苯二甲酸酯暴露的大鼠尿液生物标志物研究[J].中国环境科学,2015,35(9):2868-2873.DOI:10.3969/j.issn.1000-6923.2015.09.046.
[17]AMAGASE H,NANCE D M.A randomized,double-blind,placebocontrolled,clinical study of the general effects of a standardized Lycium barbarum(Goji)juice,GoChi[J].The Journal of Alternative and Complementary Medicine,2008,14(4):403-412.DOI:10.1089/acm.2008.0004.
[18]朱彩平.枸杞多糖的结构分析及生物活性评价[D].武汉:华中农业大学,2006:11-19.
[19]黄霞,肖丙秀,赵军伟,等.枸杞多糖对人肝癌细胞HepG2生长的影响及其分子机制[J].中草药,2010,41(9):1501-1503.
[20]如克亚·加帕尔,孙玉敬,钟烈州,等.枸杞植物化学成分及其生物活性的研究进展[J].中国食品学报,2013,13(8):161-172.DOI:10.16429/j.1009-7848.2013.08.001.
[21]SHAN Xiaozhong,ZHOU Junlai,MA Tao,et al.Lycium barbarum polysaccharides reduce exercise-induced oxidative stress[J].International Journal of Molecular Sciences,2011,12(2):1081-1088.DOI:10.3390/ijms 12021081.
[22]张盼盼.乌头碱在大鼠体内和肝微粒体的代谢及其药物相互作用的研究[D].石家庄:河北医科大学,2016:64-65.
[23]王玮,郭欣,闻福,等.孕妇尿中五种邻苯二甲酸酯代谢产物的液-液萃取反相高效液相色谱测定法[J].环境与健康杂志,2011,28(8):711-713.
[24]姜杉,郝海平,王广基.肝损状态下尿苷二磷酸葡萄糖醛酸转移酶研究进展[J].中国临床药理学与治疗学,2009,14(12):1321-1328.
[25]李晓彩.枸杞多糖对双酚A所致小鼠睾丸生精细胞凋亡的抑制作用研究[D].保定:河北农业大学,2012:21-26.
[26]蒋万志,张洪泉.枸杞多糖在免疫和抗衰老方面的研究进展[J].中国野生植物资源,2010,29(2):5-7;14.DOI:10.3969/j.issn.1006-9690.2010.02.002.
[27]XIN Yanfei,ZHOU Guoliang,DENG Zuyue,et al.Protective effect of Lycium barbarum on doxorubicin-induced cardiotoxicity[J].Phytotherapy Research,2007,21(11):1020-1024.
[28]孙小娜.枸杞多糖、槲皮素对双酚A致小鼠睾丸损伤的保护作用[D].保定:河北农业大学,2011:22-24.
[29]KOCH H M,BOLT H M,PREUSS R,et al.New metabolites of di(2-ethylhexyl)phthalate(DEHP)in human urine and serum after single oral doses of deuterium-labelled DEHP[J].Archives of Toxicology,2005,79(7):367-376.DOI:10.1007/s00204-004-0642-4.
[30]葛建,林芳,邓同乐,等.塑料中PAEs环境污染物向食品模拟物中迁移及对CYP3A4和芳香化酶的影响[J].中国食品学报,2014,14(11):165-170.DOI:10.16429/j.1009-7848.2014.11.030.
[311]COOPER B W,CHO T M,THOMPSON P M,et al.Phthalate induction of CYP3A4 is dependent on glucocorticoid regulation of PXR expression[J].Toxicological Sciences,2008,103(2):268-277.DOI:10.1093/toxsci/kfn047.
[32]周许年,毕惠嫦,金晶,等.白花前胡甲素及丙素经hCAR通路诱导UGT1A1的表达[J].药学学报,2013,48(5):794-798.DOI:10.16438/j.0513-4870.2013.05.012.
[2]PREUSS R,KOCH H M,ANGERER J.Biological monitoring of the five major metabolites of di-(2-ethylhexyl)phthalate(DEHP)in human urine using column-switching liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography B,2005,816(1/2):269-280.DOI:10.1016/j.jchromb.2004.11.048.
[3]CALAFAT A M,BROCK J W,SILVA M J,et al.Urinary and amniotic fluid levels of phthalate monoesters in rats after the oral administration of di(2-ethylhexyl)phthalate and di-n-butyl phthalate[J].Toxicology,2005,217(1):22-30.DOI:10.1016/j.tox.2005.08.013.
[4]MORTENSEN G K,MAIN K M,ANDERSSON A,et al.Determination of phthalate monoesters in human milk,consumer milk,and infant formula by tandem mass spectrometry(LC-MS-MS)[J].Analytical and Bioanalytical Chemistry,2005,382(4):1084-1092.DOI:10.1007/s00216-005-3218-0.
[5]SILVA M J,SAMANDARAMANDAR E,PREAU J L,et al.Automated solid-phase extraction and quantitative analysis of 14phthalate metabolites in human serum using isotope dilution-highperformance liquid chromatography-tandem mass spectrometry[J].Journal of Analytical Toxicology,2005,29(8):819-824.
[6]徐毅,薛山,贺稚非,等.食品包装中增塑剂DEHP毒性特点及迁移研究进展[J].食品工业科技,2013,34(21):385-389.DOI:10.13386/j.issn1002-0306.2013.21.036.
[7]DO R P,STAHLHUT R W,PONZI D,et al.Non-monotonic dose effects of in utero exposure to di(2-ethylhexyl)phthalate(DEHP)on testicular and serum testosterone and anogenital distance in male mouse fetuses[J].Reproductive Toxicology,2012,34(4):614-621.
[8]LU Yali,ZHANG Peng,LI Chenghua,et al.Characterisation of immune-related gene expression in clam(Venerupis philippinarum)under exposure to di(2-ethylhexyl)phthalate[J].Fish and Shellfish Immunology,2013,34(1):142-146.DOI:10.1016/j.fsi.2012.10.015.
[9]BIEMANN R,SANTOS A N,SANTOS A N,et al.Endocrine disrupting chemicals affect the adipogenic differentiation of mesenchymal stem cells in distinct ontogenetic windows[J].Biochemical and Biophysical Research Communications,2012,417(2):747-752.DOI:10.1016/j.bbrc.2011.12.028.
[10]杨文杰.DEHP和MEHP神经发育毒性的体外研究[D].武汉:华中科技大学,2010:48-52.DOI:10.7666/d.d153151.
[11]王丽,袁晶,张荣.邻苯二甲酸二(2-乙基己基)酯环境暴露与人群健康研究进展[J].环境与健康杂志,2009,26(5):465-467.
[12]ITO Y,YOKOTA H,WANG Ruisheng,et al.Species differences in the metabolism of di(2-ethylhexyl)phthalate(DEHP)in several organs of mice,rats,and marmosets[J].Archives of Toxicology,2005,79(3):147-154.DOI:10.1007/s00204-004-0615-7.
[13]SILVA M J,BARR D B,REIDY J A,et al.Glucuronidation patterns of common urinary and serum monoester phthalate metabolites[J].Archives of Toxicology,2003,77(10):561-567.DOI:10.1007/s00204-003-0486-3.
[14]SCHETTLER T.Human exposure to phthalates via consumer products[J].International Journal of Andrology,2006,29(1):134-139.DOI:10.1111/j.1365-2605.2005.00567.x.
[15]BURCHELL B,NEBERT D W,NELSON D R,et al.The UDP glucuronosyltransferase gene superfamily:suggested nomenclature based on evolutionary divergence[J].DNA and Cell Biology,1991,10(7):487-494.DOI:10.1089/dna.1991.10.487.
[16]柳春红,张明明,赖玉婷,等.邻苯二甲酸酯暴露的大鼠尿液生物标志物研究[J].中国环境科学,2015,35(9):2868-2873.DOI:10.3969/j.issn.1000-6923.2015.09.046.
[17]AMAGASE H,NANCE D M.A randomized,double-blind,placebocontrolled,clinical study of the general effects of a standardized Lycium barbarum(Goji)juice,GoChi[J].The Journal of Alternative and Complementary Medicine,2008,14(4):403-412.DOI:10.1089/acm.2008.0004.
[18]朱彩平.枸杞多糖的结构分析及生物活性评价[D].武汉:华中农业大学,2006:11-19.
[19]黄霞,肖丙秀,赵军伟,等.枸杞多糖对人肝癌细胞HepG2生长的影响及其分子机制[J].中草药,2010,41(9):1501-1503.
[20]如克亚·加帕尔,孙玉敬,钟烈州,等.枸杞植物化学成分及其生物活性的研究进展[J].中国食品学报,2013,13(8):161-172.DOI:10.16429/j.1009-7848.2013.08.001.
[21]SHAN Xiaozhong,ZHOU Junlai,MA Tao,et al.Lycium barbarum polysaccharides reduce exercise-induced oxidative stress[J].International Journal of Molecular Sciences,2011,12(2):1081-1088.DOI:10.3390/ijms 12021081.
[22]张盼盼.乌头碱在大鼠体内和肝微粒体的代谢及其药物相互作用的研究[D].石家庄:河北医科大学,2016:64-65.
[23]王玮,郭欣,闻福,等.孕妇尿中五种邻苯二甲酸酯代谢产物的液-液萃取反相高效液相色谱测定法[J].环境与健康杂志,2011,28(8):711-713.
[24]姜杉,郝海平,王广基.肝损状态下尿苷二磷酸葡萄糖醛酸转移酶研究进展[J].中国临床药理学与治疗学,2009,14(12):1321-1328.
[25]李晓彩.枸杞多糖对双酚A所致小鼠睾丸生精细胞凋亡的抑制作用研究[D].保定:河北农业大学,2012:21-26.
[26]蒋万志,张洪泉.枸杞多糖在免疫和抗衰老方面的研究进展[J].中国野生植物资源,2010,29(2):5-7;14.DOI:10.3969/j.issn.1006-9690.2010.02.002.
[27]XIN Yanfei,ZHOU Guoliang,DENG Zuyue,et al.Protective effect of Lycium barbarum on doxorubicin-induced cardiotoxicity[J].Phytotherapy Research,2007,21(11):1020-1024.
[28]孙小娜.枸杞多糖、槲皮素对双酚A致小鼠睾丸损伤的保护作用[D].保定:河北农业大学,2011:22-24.
[29]KOCH H M,BOLT H M,PREUSS R,et al.New metabolites of di(2-ethylhexyl)phthalate(DEHP)in human urine and serum after single oral doses of deuterium-labelled DEHP[J].Archives of Toxicology,2005,79(7):367-376.DOI:10.1007/s00204-004-0642-4.
[30]葛建,林芳,邓同乐,等.塑料中PAEs环境污染物向食品模拟物中迁移及对CYP3A4和芳香化酶的影响[J].中国食品学报,2014,14(11):165-170.DOI:10.16429/j.1009-7848.2014.11.030.
[311]COOPER B W,CHO T M,THOMPSON P M,et al.Phthalate induction of CYP3A4 is dependent on glucocorticoid regulation of PXR expression[J].Toxicological Sciences,2008,103(2):268-277.DOI:10.1093/toxsci/kfn047.
[32]周许年,毕惠嫦,金晶,等.白花前胡甲素及丙素经hCAR通路诱导UGT1A1的表达[J].药学学报,2013,48(5):794-798.DOI:10.16438/j.0513-4870.2013.05.012.