羧甲基赖氨酸在方便面中含量及其对雄性生殖和胚胎发育毒性的研究
|
摘要
采用酶联免疫吸附测定法(ELISA)对市售20个品牌方便面中游离态和结合态羧甲基赖氨酸(CML)含量进行了定量分析,并以体外细胞培养评价了CML的雄性生殖毒性和胚胎发育毒性。市售方便面中,游离态CML质量分数为0.39~0.80 mg/kg,结合态CML质量分数为19.42~34.99 mg/kg。CML作用于睾丸间质细胞R2C细胞24 h,浓度在1 mmol/L以下对细胞存活率没有显著抑制作用,浓度在2 mmol/L以下对孕酮分泌量没有显著的抑制作用;CML作用于胚胎干(ES)细胞D3和成纤维细胞3T3 10 d,浓度在4 mmol/L以下对3T3细胞存活率没有显著影响,浓度在0.5 mmol/L以下对D3细胞存活率没有显著影响。根据市售方便面中的CML浓度评估CML的摄入量,常规食用方便面摄入的CML不会对这几种细胞的存活率和R2C细胞的孕酮分泌量产生显著抑制作用,可认为不会产生雄性生殖和胚胎发育毒性。
Nε-(carboxymethyl) lysine(CML) concentrations in instant noodles of 20 brands were determinated using enzyme-linked immunosorbent assay(ELISA),then developmental and reproductive toxicity of CML were evaluated in vitro. Concentrations of CML in free form and bind form were 0.39~0.80 mg/kg and 19.42~34.99 mg/kg,respectively. No significant effect of CML was found on cell viability(CML≤1 mmol/L) and progesterone secretion(CML≤2 mmol/L) of R2 C cells treated with CML for 24 h. Moreover,CML of concentration lower than 4 mmol/L has no significant effect on 3 T3 cells treated with CML for 10 days,and CML of concentration lower than0.5 mmol/L has no significant effect on D3 cells treated with CML for 10 days. According to the concentrations of CML detected in the present study,CML would not inhibit the progesterone secretion of R2 C cells or cell viability of R2 C,3 T3,and D3 cells with regular intake of instant noodles. Thus,regular intake of instant noodles would not cause developmental or male reproductive toxicity induced by CML.
Nε-(carboxymethyl) lysine(CML) concentrations in instant noodles of 20 brands were determinated using enzyme-linked immunosorbent assay(ELISA),then developmental and reproductive toxicity of CML were evaluated in vitro. Concentrations of CML in free form and bind form were 0.39~0.80 mg/kg and 19.42~34.99 mg/kg,respectively. No significant effect of CML was found on cell viability(CML≤1 mmol/L) and progesterone secretion(CML≤2 mmol/L) of R2 C cells treated with CML for 24 h. Moreover,CML of concentration lower than 4 mmol/L has no significant effect on 3 T3 cells treated with CML for 10 days,and CML of concentration lower than0.5 mmol/L has no significant effect on D3 cells treated with CML for 10 days. According to the concentrations of CML detected in the present study,CML would not inhibit the progesterone secretion of R2 C cells or cell viability of R2 C,3 T3,and D3 cells with regular intake of instant noodles. Thus,regular intake of instant noodles would not cause developmental or male reproductive toxicity induced by CML.
引文
[1]SINGH R,BARDEN A,MORI T,et al.Advanced glycation end-products:a review[J].Diabetologia,2001,44(2):129-146.
[2]XANTHIS A,HATZITOLIOS A,KOLIAKOS G,et al.Advanced glycosylation end products and nutrition-a possible relation with diabetic atherosclerosis and how to prevent it[J].Journal of Food Science,2007,72(8):R125-129.
[3]HENLE T.Dietary advanced glycation end products--a risk to human health?A call for an interdisciplinary debate[J].Molecular Nutrition&Food Research,2007,51(9):1075-1078.
[4]SEBEKOVA K,SOMOZA V.Dietary advanced glycation endproducts(AGEs)and their health effects-PRO[J].Molecular Nutrition&Food Research,2007,51(9):1079-1084.
[5]LIU Yixiang,JING Hao.Advances in pathological effects of maillard reaction products in vivo[J].Journal of Food Science and Biotechnology,2010,29(2):161-166.(in Chinese)
[6]THORPE R,BAYNES W.CML a brief history[J].International Congress Series,2002,1245:91-99.
[7]ZHANG Z,SUN J,BAI W,et al.Advances in toxicology of Nε-(carboxymethyl)-lysine(CML)[J].Agricultural Science&Technology,2013,14(10):1403-1408.
[8]CAI W,HE J C,ZHU L,et al.High levels of dietary advanced glycation end products transform low-density lipoprotein into a potent redox-sensitive mitogen-activated protein kinase stimulant in diabetic patients[J].Circulation,2004,110(3):285-291.
[9]GONG Z,GUANGWEI H,LU X,et al.Determination of advanced glycation endproducts by LC-MS/MS in raw and roasted almonds(Prunus dulcis)[J].Journal of Agricultural&Food Chemistry,2011,59(22):12037-12046.
[10]TAREKE E,FORSLUND A,LINDH C H,et al.Isotope dilution ESI-LC-MS/MS for quantification of free and total Nepsilon-(1-Carboxymethyl)-L-Lysine and free Nepsilon-(1-Carboxyethyl)-L-Lysine:comparison of total Nepsilon-(1-Carboxymethyl)-L-Lysine levels measured with new method to ELISA assay in gruel samples[J].Food Chemistry,2013,141(4):4253-4259.
[11]CHARISSOU A,AIT-AMEUR L,BIRLOUEZ A I.Evaluation of a gas chromatography/mass spectrometry method for the quantification of carboxymethyllysine in food samples[J].Journal of Chromatography A,2007,1140(1-2):189-94.
[12]LOAEC G,JACOLOT P,HELOU C,et al.Acrylamide,5-hydroxymethylfurfural and N(epsilon)-carboxymethyl-lysine in coffee substitutes and instant coffees[J].Food Additives&Contaminants Part A,Chemistry,Analysis,Control,Exposure&Risk Assessment,2014,31(4):593-604.
[13]URIBARRI J,WOODRUFF S,GOODMAN S,et al.Advanced glycation end products in foods and a practical guide to their reduction in the diet[J].Journal of the American Dietetic Association,2010,110(6):911-16.e12.
[14]周燕琼.植物多酚抑制食品中晚期糖基化终末产物的形成的作用机理研究[D].杭州:浙江大学,2015.
[15]DELGADO A C,TESSIER F J,NIQUET L C,et al.Study of the urinary and faecal excretion of Nepsilon-carboxymethyllysine in young human volunteers[J].Amino Acids,2012,43(2):595-602.
[16]BEATTIE M C,ADEKOLA L,PAPADOPOULOS V,et al.Leydig cell aging and hypogonadism[J].Experimental Gerontology,2015,68:87-91.
[17]ZHANG Q,ZOU P,ZHAN H,et al.Dihydrolipoamide dehydrogenase and c AMP are associated with cadmium-mediated Leydig cell damage[J].Toxicology Letters,2011,205(2):183-189.
[18]SUN J,BAI S,BAI W,et al.Toxic mechanisms of 3-monochloropropane-1,2-diol on progesterone production in R2C rat leydig cells[J].Journal of Agricultural and Food Chemistry,2013,61(41):9955-9960.
[2]XANTHIS A,HATZITOLIOS A,KOLIAKOS G,et al.Advanced glycosylation end products and nutrition-a possible relation with diabetic atherosclerosis and how to prevent it[J].Journal of Food Science,2007,72(8):R125-129.
[3]HENLE T.Dietary advanced glycation end products--a risk to human health?A call for an interdisciplinary debate[J].Molecular Nutrition&Food Research,2007,51(9):1075-1078.
[4]SEBEKOVA K,SOMOZA V.Dietary advanced glycation endproducts(AGEs)and their health effects-PRO[J].Molecular Nutrition&Food Research,2007,51(9):1079-1084.
[5]LIU Yixiang,JING Hao.Advances in pathological effects of maillard reaction products in vivo[J].Journal of Food Science and Biotechnology,2010,29(2):161-166.(in Chinese)
[6]THORPE R,BAYNES W.CML a brief history[J].International Congress Series,2002,1245:91-99.
[7]ZHANG Z,SUN J,BAI W,et al.Advances in toxicology of Nε-(carboxymethyl)-lysine(CML)[J].Agricultural Science&Technology,2013,14(10):1403-1408.
[8]CAI W,HE J C,ZHU L,et al.High levels of dietary advanced glycation end products transform low-density lipoprotein into a potent redox-sensitive mitogen-activated protein kinase stimulant in diabetic patients[J].Circulation,2004,110(3):285-291.
[9]GONG Z,GUANGWEI H,LU X,et al.Determination of advanced glycation endproducts by LC-MS/MS in raw and roasted almonds(Prunus dulcis)[J].Journal of Agricultural&Food Chemistry,2011,59(22):12037-12046.
[10]TAREKE E,FORSLUND A,LINDH C H,et al.Isotope dilution ESI-LC-MS/MS for quantification of free and total Nepsilon-(1-Carboxymethyl)-L-Lysine and free Nepsilon-(1-Carboxyethyl)-L-Lysine:comparison of total Nepsilon-(1-Carboxymethyl)-L-Lysine levels measured with new method to ELISA assay in gruel samples[J].Food Chemistry,2013,141(4):4253-4259.
[11]CHARISSOU A,AIT-AMEUR L,BIRLOUEZ A I.Evaluation of a gas chromatography/mass spectrometry method for the quantification of carboxymethyllysine in food samples[J].Journal of Chromatography A,2007,1140(1-2):189-94.
[12]LOAEC G,JACOLOT P,HELOU C,et al.Acrylamide,5-hydroxymethylfurfural and N(epsilon)-carboxymethyl-lysine in coffee substitutes and instant coffees[J].Food Additives&Contaminants Part A,Chemistry,Analysis,Control,Exposure&Risk Assessment,2014,31(4):593-604.
[13]URIBARRI J,WOODRUFF S,GOODMAN S,et al.Advanced glycation end products in foods and a practical guide to their reduction in the diet[J].Journal of the American Dietetic Association,2010,110(6):911-16.e12.
[14]周燕琼.植物多酚抑制食品中晚期糖基化终末产物的形成的作用机理研究[D].杭州:浙江大学,2015.
[15]DELGADO A C,TESSIER F J,NIQUET L C,et al.Study of the urinary and faecal excretion of Nepsilon-carboxymethyllysine in young human volunteers[J].Amino Acids,2012,43(2):595-602.
[16]BEATTIE M C,ADEKOLA L,PAPADOPOULOS V,et al.Leydig cell aging and hypogonadism[J].Experimental Gerontology,2015,68:87-91.
[17]ZHANG Q,ZOU P,ZHAN H,et al.Dihydrolipoamide dehydrogenase and c AMP are associated with cadmium-mediated Leydig cell damage[J].Toxicology Letters,2011,205(2):183-189.
[18]SUN J,BAI S,BAI W,et al.Toxic mechanisms of 3-monochloropropane-1,2-diol on progesterone production in R2C rat leydig cells[J].Journal of Agricultural and Food Chemistry,2013,61(41):9955-9960.