可溶性大豆多糖改善左旋肉碱诱导的小鼠小肠首过代谢
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摘要
目的:研究大豆可溶性多糖(soybean soluble polysaccharides,SSPS)对左旋肉碱喂养小鼠的小肠代谢、氧化应激和炎症相关蛋白表达的影响。方法:24只雄性昆明小鼠随机分成正常组、左旋肉碱组、SSPS组3个实验组。连续饲喂56 d后处死小鼠,测定小鼠小肠内P-糖蛋白(P-glycoprotein,P-gp)、多药耐药性蛋白(multidrug resistanceproteins,MRP)1、MRP3、白细胞介素(interleukin,IL)-1、IL-6、肿瘤坏死因子-α(tumornecrosis factor-α,TNF-α)质量浓度和尿苷二磷酸葡萄糖醛酸基转移酶(uridine diphosphate glucuronosyltransferase,UGT)活力、硫酸基转移酶(sulfotransferase,SULT)活力、丙二醛(malondialdehyde,MDA)浓度等的变化。结果:与正常组相比,左旋肉碱组小鼠小肠具有更高的P-gp和MRP3质量浓度(P<0.05);而SSPS可使左旋肉碱喂养小鼠小肠内P-gp质量浓度显著降低(P<0.05),且使MRP3质量浓度轻微下降,表明SSPS可有效抑制左旋肉碱诱导的首过代谢。此外,SSPS与左旋肉碱联合处理可有效避免小鼠小肠组织MDA浓度的增加、超氧化物歧化酶活力的下降和·OH清除能力的减弱,预示SSPS可抑制长期摄入左旋肉碱诱导的氧化应激。然而,所有小鼠小肠组织的SULT和UGT活力以及IL-1、IL-6、TNF-α质量浓度没有显著变化(P>0.05),表明过量左旋肉碱和SSPS处理不会影响II相代谢,也不会导致炎症反应。此外,代谢组学分析表明,SSPS可抑制长期过量摄入左旋肉碱导致的能量代谢及脂质代谢紊乱。结论:SSPS可以通过调控氧化应激和代谢紊乱而有效预防左旋肉碱诱导的小鼠小肠首过代谢。
Objective:To study the effect of soluble soybean polysaccharides(SSPS) on intestinal metabolism,oxidative stress and inflammation-related proteins in mice fed L-carnitine.Methods:A total of 24 male Kunming mice were randomly divided into three groups:normal group,L-carnitine group and SSPS group.At the end of the 56-day feeding period,all mice were sacrificed to assess the changes in the levels of P-glycoprotein(P-gp),multidrug resistance protein(MRP) 1,MRP3,interleukin(IL)-1,IL-6,tumor necrosis factor-α(TNF-α),uridine diphosphateglucuronosyltransferase(UGT),sulfotransferase(SULT),and malondialdehyde(MDA) in the small intestine.Results:The concentrations of P-gp and MRP3 in the small intestine homogenate supernatant of mice in the L-carnitine group were significantly higher than that of the normal group(P<0.05).However,SSPS significantly decreased P-gp concentration(P<0.05),and slightly decreased MRP3 concentration in the small intestine of L-carnitine-fed mice,suggesting that SSPS could inhibit L-carnitine-induced first pass metabolism.Additionally,co-treatment of SSPS and L-carnitine on mice effectively prevented the increase of MDA content,the decrease of superoxide dismutase(SOD) activity,and the weakening of hydroxyl radical-scavenging capacity,indicating that SSPS could inhibit L-carnitine-induced oxidative stress.However,the activities of SULT and UGT,and the concentrations of IL-1,IL-6 and TNF-α in the small intestine did not differ significantly among all groups of mice(P>0.05),suggesting that excessive L-carnitine and SSPS treatment did not affect the II phase metabolism or lead to inflammation.Furthermore,metabonomic analysis showed that SSPS could inhibit the disorders of energy metabolism and lipid metabolism caused by long-term excessive intake of L-carnitine.Conclusion:The first pass metabolism induced by L-carnitine can be effectively prevented by SSPS via controlling oxidative stress and metabolic disorders.
Objective:To study the effect of soluble soybean polysaccharides(SSPS) on intestinal metabolism,oxidative stress and inflammation-related proteins in mice fed L-carnitine.Methods:A total of 24 male Kunming mice were randomly divided into three groups:normal group,L-carnitine group and SSPS group.At the end of the 56-day feeding period,all mice were sacrificed to assess the changes in the levels of P-glycoprotein(P-gp),multidrug resistance protein(MRP) 1,MRP3,interleukin(IL)-1,IL-6,tumor necrosis factor-α(TNF-α),uridine diphosphateglucuronosyltransferase(UGT),sulfotransferase(SULT),and malondialdehyde(MDA) in the small intestine.Results:The concentrations of P-gp and MRP3 in the small intestine homogenate supernatant of mice in the L-carnitine group were significantly higher than that of the normal group(P<0.05).However,SSPS significantly decreased P-gp concentration(P<0.05),and slightly decreased MRP3 concentration in the small intestine of L-carnitine-fed mice,suggesting that SSPS could inhibit L-carnitine-induced first pass metabolism.Additionally,co-treatment of SSPS and L-carnitine on mice effectively prevented the increase of MDA content,the decrease of superoxide dismutase(SOD) activity,and the weakening of hydroxyl radical-scavenging capacity,indicating that SSPS could inhibit L-carnitine-induced oxidative stress.However,the activities of SULT and UGT,and the concentrations of IL-1,IL-6 and TNF-α in the small intestine did not differ significantly among all groups of mice(P>0.05),suggesting that excessive L-carnitine and SSPS treatment did not affect the II phase metabolism or lead to inflammation.Furthermore,metabonomic analysis showed that SSPS could inhibit the disorders of energy metabolism and lipid metabolism caused by long-term excessive intake of L-carnitine.Conclusion:The first pass metabolism induced by L-carnitine can be effectively prevented by SSPS via controlling oxidative stress and metabolic disorders.
引文
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[21]杨晓惠.左旋肉碱与降脂减肥[J].生物技术世界,2015(4):129-130.
[22]刘磊.长期口服过量左旋肉碱对健康成年SD大鼠正常代谢的干扰作用[D].南京:南京大学,2012:22-45.
[23]GUO J,MENG Y,ZHAO Y,et al.Myricetin derived from Hovenia dulcis Thunb.ameliorates vascular endothelial dysfunction and liver injury in high choline-fed mice[J].Food&Function,2015,6(5):1620-1634.DOI:10.1039/c4fo01073f.
[24]ROLO A P,TEODORO J S,PALMEIRA C M.Role of oxidative stress in the pathogenesis of nonalcoholic steatohepatitis[J].Free Radical Biology&Medicine,2012,52(1):59-69.DOI:10.1016/j.freeradbiomed.2011.10.003.
[25]MARCHETTI S,M AZZANTI R,BEIJNEN J H,et al.Concise review:clinical relevance of drug drug and herb drug interactions media ted by the ABC transporter ABCB1(MDR1,P-glycoprotein)[J].The Oncologist,2007,12(8):927-941.DOI:10.1634/theoncologist.12-8-927.
[26]SUZUKI H,SUGIYAMA Y.Role of metabolic enzymesand efflux transporters in the absorption of drugs from the small intestine[J].European Journal of Pharmaceutical Sciences,2000,12(1):3-12.DOI:10.1016/S0928-0987(00)00178-0.
[27]毛晓琴,谢海棠,周宏灏.多药耐药蛋白和CYP3A4介导的中草药-西药相互作用[J].中国临床药理学与治疗学,2007,12(7):728-734.DOI:10.3969/j.issn.1009-2501.2007.07.002.
[28]LI W F,LU Y L,HUANG D,et al.Effects of stachyose on absorptionand transportation of tea catechins in mice:possible role of phase II metabolic enzymes and efflux transporters inhibition by stachyose[J].Food&Nutrition Research,2016,60(1):32783.DOI:10.3402/fnr.v60.32783.
[29]李利,赵远胜,杜飞飞,等.标准银杏叶提取物GBE50中多种活性成分的肠道吸收和系统前处置研究[C]//中药与天然药高峰论坛暨第十二届全国中药和天然药物学术研讨会.北京:中国药学会,2012:444.
[30]LI W,LI Z,HAN X,et al.Enhancing the hepatic protective effect of genistein by oral administration with stachyose in mice with chronic high fructose diet consumption[J].Food&Function,2016,7(5):2420-2430.DOI:10.1039/c6fo00038j.
[31]罗霞,罗爽,郑彦懿,等.肠道菌群失调增加小鼠肠上皮内T淋巴细胞活化和促炎细胞因子分泌[J].细胞与分子免疫学杂志,2016,32(8):1031-1035.DOI:10.13423/j.cnki.cjcmi.007812.
[2]LIU L,ZHANG D M,WANG M X,et al.The adverse effects of long-term L-carnitine supplementation on liver and kidney function in rats[J].Human and Experimental Toxicology,2015,34(11):1148-1161.DOI:10.1177/0960327115571767.
[3]仝晨均,付晓丽,付余,等.左旋肉碱对肥胖患者血脂水平影响的meta分析[J].郑州大学学报(医学版),2015,50(2):214-217.DOI:10.13705/j.issn.1671-6825.2015.02.018.
[4]张晓红,杜玲,顾伟军,等.左旋肉碱对脂肪肝患者的干预效果[J].温州医科大学学报,2015,45(1):61-64.DOI:10.3969/j.issn.2095-9400.2015.01.015.
[5]KOETH R A,WANG Z,LEVISON B S,et al.Intestinal microbiota metabolism of L-carnitine,a nutrient in red meat,promotes atherosclerosis[J].Nature Medicine,2013,19(5):576-585.DOI:10.1038/nm.3145.
[6]LI W,ZHANG R,GUO J,et al.Protective effect of R.glutinosa oligosaccharides against high L-carnitine diet-induced endothelial dysfunction and hepatic injury in mice[J].International Journal of Biological Macromolecules,2016,85:285-293.DOI:10.1016/j.ijbiomac.2015.12.092.
[7]GREWAL G K,KUKAL S,KANOJIA N,et al.Effect of oxidative stress on ABC transporters:contribution to epilepsy pharmacoresistance[J].Molecules,2017,22(3):e365.DOI:10.3390/molecules22030365.
[8]邓婧婷,庄笑梅,李桦.小肠首过代谢的研究进展[J].国际药学研究杂志,2010,37(4):269-272.DOI:10.13220/j.cnki.jipr.2010.04.011.
[9]BRAND W,SCHUTTE M E,WILLIAMSON G,et al.Flavonoidmediated inhibition of intestinal ABC transporters may affect the oral bioavailability of drugs,food-borne toxic compounds and bioactive ingredients[J].Biomedicine&Pharmacotherapy,2006,60(9):508-519.DOI:10.1016/j.biopha.2006.07.081.
[10]TIAN Q E.Astragalus polysaccharides can regulate cytokine and P-glycoprotein expression in H22 tumor-bearing mice[J].World Journal of Gastroenterology,2012,18(47):7079-7086.DOI:10.3748/wjg.v18.i47.7079.
[11]LI W D,ZHANG B D,WEI R,et al.Reversal effect of Ganoderma lucidum polysaccharide on multidrug resistance in K562/ADMcell line[J].Acta Pharmacologica Sinica,2008,29(5):620-627.DOI:10.1111/j.1745-7254.2008.00776.x.
[12]NAKAMURA A,FURUTA H,MAEDA H,et al.Structural studies by stepwise enzymatic degradation of the main backbone of soybean soluble polysaccharides consisting of galacturonan and rhamnogalacturonan[J].Bioscience,Biotechnology,and Biochemistry,2002,66(6):1301-1313.DOI:10.1271/bbb.66.1301.
[13]SUN X Y,OUYANG J M,BHADJA P,et al.Protective effects of degraded soybean polysaccharides on renal epithelial cells exposed to oxidative damage[J].Journal of Agricultural and Food Chemistry,2016,64(42):7911-7920.DOI:10.1021/acs.jafc.6b03323.
[14]樊星,翁谢川,王和枚,等.维生素D和维生素E在DSS诱导大鼠急性IBD模型中的作用及机制[J].中国药理学与毒理学杂志,2013,27(3):465.
[15]梁帅,王晗,陈珺,等.氢化豆油摄入对小鼠小肠反式脂肪酸含量及IL-8表达的影响[J].大连工业大学学报,2018(4):239-243.DOI:10.19670/j.cnki.dlgydxxb.2018.0402.
[16]ZENG J,YIN P,TAN Y,et al.Metabolomics study of hepatocellular carcinoma:discovery and validation of serum potential biomarkers by using capillary electrophoresis-mass spectrometry[J].Journal of Proteome Research,2014,13(7):3420-3431.DOI:10.1021/pr500390y.
[17]TAN G G,LOU Z Y,LIAO W T,et al.Potential biomarkers in mouse myocardium of doxorubicin-induced cardiomyopathy:a metabonomic method and its application[J].PLoS ONE,2011,6(11):e27683.DOI:10.1371/journal.pone.0027683.
[18]HUANG S M,TOH W,BENKE P I,et al.MetaboNexus:an interactive platform for integrated metabolomics analysis[J].Metabolomics,2014,10(6):1084-1093.DOI:10.1007/s11306-014-0648-8.
[19]LIQUET B,LE CAO K A,HOCINI H,et al.A novel approach for biomarker selection and the integration of repeated measures experiments from two assays[J].BMC Bioinformatics,2012,13:325.DOI:10.1186/1471-2105-13-325.
[20]SREEKUMAR A,POISSON L M,RAJENDIRAN T M,et al.Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression[J].Nature,2009,457:910-914.DOI:10.1038/nature07762.
[21]杨晓惠.左旋肉碱与降脂减肥[J].生物技术世界,2015(4):129-130.
[22]刘磊.长期口服过量左旋肉碱对健康成年SD大鼠正常代谢的干扰作用[D].南京:南京大学,2012:22-45.
[23]GUO J,MENG Y,ZHAO Y,et al.Myricetin derived from Hovenia dulcis Thunb.ameliorates vascular endothelial dysfunction and liver injury in high choline-fed mice[J].Food&Function,2015,6(5):1620-1634.DOI:10.1039/c4fo01073f.
[24]ROLO A P,TEODORO J S,PALMEIRA C M.Role of oxidative stress in the pathogenesis of nonalcoholic steatohepatitis[J].Free Radical Biology&Medicine,2012,52(1):59-69.DOI:10.1016/j.freeradbiomed.2011.10.003.
[25]MARCHETTI S,M AZZANTI R,BEIJNEN J H,et al.Concise review:clinical relevance of drug drug and herb drug interactions media ted by the ABC transporter ABCB1(MDR1,P-glycoprotein)[J].The Oncologist,2007,12(8):927-941.DOI:10.1634/theoncologist.12-8-927.
[26]SUZUKI H,SUGIYAMA Y.Role of metabolic enzymesand efflux transporters in the absorption of drugs from the small intestine[J].European Journal of Pharmaceutical Sciences,2000,12(1):3-12.DOI:10.1016/S0928-0987(00)00178-0.
[27]毛晓琴,谢海棠,周宏灏.多药耐药蛋白和CYP3A4介导的中草药-西药相互作用[J].中国临床药理学与治疗学,2007,12(7):728-734.DOI:10.3969/j.issn.1009-2501.2007.07.002.
[28]LI W F,LU Y L,HUANG D,et al.Effects of stachyose on absorptionand transportation of tea catechins in mice:possible role of phase II metabolic enzymes and efflux transporters inhibition by stachyose[J].Food&Nutrition Research,2016,60(1):32783.DOI:10.3402/fnr.v60.32783.
[29]李利,赵远胜,杜飞飞,等.标准银杏叶提取物GBE50中多种活性成分的肠道吸收和系统前处置研究[C]//中药与天然药高峰论坛暨第十二届全国中药和天然药物学术研讨会.北京:中国药学会,2012:444.
[30]LI W,LI Z,HAN X,et al.Enhancing the hepatic protective effect of genistein by oral administration with stachyose in mice with chronic high fructose diet consumption[J].Food&Function,2016,7(5):2420-2430.DOI:10.1039/c6fo00038j.
[31]罗霞,罗爽,郑彦懿,等.肠道菌群失调增加小鼠肠上皮内T淋巴细胞活化和促炎细胞因子分泌[J].细胞与分子免疫学杂志,2016,32(8):1031-1035.DOI:10.13423/j.cnki.cjcmi.007812.