氧化应激对蛋氨酸负载后大鼠肝细胞同型半胱氨酸代谢的影响
양화응격대단안산부재후대서간세포동형반광안산대사적영향
Effects of oxidative stress on homocysteine metabolism in methionine loading rat hepatocytes
  • 万方数据
    中华临床营养杂志 중화림상영양잡지
    CHINESE JOURNAL OF CLINICAL NUTRITION
    2012年 4期 238-243 ,共6页

    韩超%高蔚娜%吴健全%韦京豫%蒲玲玲%杨继军%郭长江

    한초%고위나%오건전%위경예%포령령%양계군%곽장강

    蛋氨酸负载%同型半胱氨酸%氧化应激 蛋氨痠負載%同型半胱氨痠%氧化應激
    단안산부재%동형반광안산%양화응격
    Methionine loading%Homocysteine%Oxidative stress
    目的 研究氧化应激对蛋氨酸负载后BRL大鼠肝细胞同型半胱氨酸及相关氨基酸代谢的影响.方法 体外培养BRL大鼠肝细胞,将大鼠肝细胞分为对照组、氧化应激组(100 μmol/L H2O2作用2 h)、蛋氨酸组(50 mmol/L蛋氨酸作用1h)、氧化应激±蛋氨酸组(100 μmol/L H2O2作用2h+50 mmol/L蛋氨酸作用1h),实验结束收集各组培养液上清.采用高效液相法测定同型半胱氨酸、半胱氨酸和谷胱甘肽的含量,采用全自动氨基酸分析仪测定相关氨基酸的含量.结果 与对照组比较,蛋氨酸组同型半胱氨酸[(3.76±0.22)比(1.54±0.05)μmol/L,P=0.000]、半胱氨酸[(199.80±8.75)比(99.11 ±2.47)μmol/L,P=0.000]的含量显著增加,氧化应激+蛋氨酸组同型半胱氨酸[(3.84±0.34)比(1.54±0.05)μmol/L,P=0.000]、半胱氨酸[(200.66±8.60)比(99.11±2.47)μmoL/L,P=0.000]的含量显著增加.与氧化应激组比较,蛋氨酸组同型半胱氨酸[(3.76±0.22)比(1.67±0.13)μmol/L,P=0.000]、半胱氨酸[(199.80±8.75)比(82.64±15.88)μmol/L,P=0.000]、谷胱甘肽[(1.50±0.14)比(1.00±0.11)μmol/L,P=0.011]含量均显著增加,H2 O2+蛋氨酸组的同型半胱氨酸[(3.84±0.34)比(1.67±0.13)μmol/L,P=0.000]、半胱氨酸[(200.66±8.60)比(82.64±15.88)μmol/L,P=0.000]、谷胱甘肽[(1.40±0.30)比(1.00±0.11)μmoL/L,P =0.028)]含量均显著增加.氧化应激+蛋氨酸组Hcy含量较蛋氨酸组有增加的趋势,但差异无统计学意义(P =0.628).与对照组比较,氧化应激组丝氨酸[(12.41±1.51)比(24.00±2.54)mg/L,P =0.000]、谷氨酸[(33.31±0.17)比(43.10±0.52)mg/L,P=0.000]和甘氨酸[(6.23±0.18)比(24.66±10.87)mg/L,P=0.003]的含量均显著降低,而牛磺酸的含量显著增加[(7.99±0.16)比(6.17±0.15)mg/L,P=0.000];与氧化应激组比较,蛋氨酸组丝氨酸[(16.98±0.39)比(12.41±1.51)mg/L,P=0.006]和谷氨酸[(35.44±0.82)比(33.31±0.17)mg/L,P=0.002]含量显著增加,而牛磺酸含量[(3.77±0.16)比(7.99±0.16)mg/L,P=0.000]显著减少;与蛋氨酸组比较,氧化应激+蛋氨酸组丝氨酸[(12.59±0.66)比(16.98±0.39)mg/L,P=0.008]、谷氨酸[(30.87±0.60)比(35.44±0.82)mg/L,P=0.000]的含量均显著降低,而牛磺酸的含量[(4.37±0.12)比(3.77±0.16)mg/L,P=0.001]显著增加.结论 氧化应激对蛋氨酸负载后BRL大鼠肝细胞的同型半胱氨酸代谢可能具有一定的促进作用,不过主要体现的是蛋氨酸负载的影响.
    목적 연구양화응격대단안산부재후BRL대서간세포동형반광안산급상관안기산대사적영향.방법 체외배양BRL대서간세포,장대서간세포분위대조조、양화응격조(100 μmol/L H2O2작용2 h)、단안산조(50 mmol/L단안산작용1h)、양화응격±단안산조(100 μmol/L H2O2작용2h+50 mmol/L단안산작용1h),실험결속수집각조배양액상청.채용고효액상법측정동형반광안산、반광안산화곡광감태적함량,채용전자동안기산분석의측정상관안기산적함량.결과 여대조조비교,단안산조동형반광안산[(3.76±0.22)비(1.54±0.05)μmol/L,P=0.000]、반광안산[(199.80±8.75)비(99.11 ±2.47)μmol/L,P=0.000]적함량현저증가,양화응격+단안산조동형반광안산[(3.84±0.34)비(1.54±0.05)μmol/L,P=0.000]、반광안산[(200.66±8.60)비(99.11±2.47)μmoL/L,P=0.000]적함량현저증가.여양화응격조비교,단안산조동형반광안산[(3.76±0.22)비(1.67±0.13)μmol/L,P=0.000]、반광안산[(199.80±8.75)비(82.64±15.88)μmol/L,P=0.000]、곡광감태[(1.50±0.14)비(1.00±0.11)μmol/L,P=0.011]함량균현저증가,H2 O2+단안산조적동형반광안산[(3.84±0.34)비(1.67±0.13)μmol/L,P=0.000]、반광안산[(200.66±8.60)비(82.64±15.88)μmol/L,P=0.000]、곡광감태[(1.40±0.30)비(1.00±0.11)μmoL/L,P =0.028)]함량균현저증가.양화응격+단안산조Hcy함량교단안산조유증가적추세,단차이무통계학의의(P =0.628).여대조조비교,양화응격조사안산[(12.41±1.51)비(24.00±2.54)mg/L,P =0.000]、곡안산[(33.31±0.17)비(43.10±0.52)mg/L,P=0.000]화감안산[(6.23±0.18)비(24.66±10.87)mg/L,P=0.003]적함량균현저강저,이우광산적함량현저증가[(7.99±0.16)비(6.17±0.15)mg/L,P=0.000];여양화응격조비교,단안산조사안산[(16.98±0.39)비(12.41±1.51)mg/L,P=0.006]화곡안산[(35.44±0.82)비(33.31±0.17)mg/L,P=0.002]함량현저증가,이우광산함량[(3.77±0.16)비(7.99±0.16)mg/L,P=0.000]현저감소;여단안산조비교,양화응격+단안산조사안산[(12.59±0.66)비(16.98±0.39)mg/L,P=0.008]、곡안산[(30.87±0.60)비(35.44±0.82)mg/L,P=0.000]적함량균현저강저,이우광산적함량[(4.37±0.12)비(3.77±0.16)mg/L,P=0.001]현저증가.결론 양화응격대단안산부재후BRL대서간세포적동형반광안산대사가능구유일정적촉진작용,불과주요체현적시단안산부재적영향.
    Objective To investigate the effects of oxidative stress on homocysteine and related amino acids metabolism in methionine-loading BRL rat hepatocytes.Methods Cultured BRL rat hepatocytes were divided into control and oxidatively stressed group(100 μmol/L H2O2 was added in culture medium for 2 hours),methionine group(50 mmol/L methionine was added in culture medium for 1 hour),and oxidatively stressed + methionine group(100 μmol/L H2O2 was added in culture medium for 2 hours + 50 mmol/L methionine was added in culture medium for 1 hour).At the end of the experiment,culture fluid was collected.Homocysteine,cysteine,and glutathione were measured by high-performance liquid chromatography,and amino acids were assayed by amino acids analyzer.Results Compared with the control group,the contents of homocysteine[(3.76 ± 0.22)vs.(1.54±0.05)μmol/L,P=0.000]and cysteine[(199.80 ±8.75)vs.(99.11 ±2.47)μmol/L,P=0.000]significantly increased in methionine group,and the contents of homocysteine[(3.84 ± 0.34)vs.(1.54 ±0.05)μmol/L,P=0.000]and cysteine[(200.66±8.60)vs.(99.11 ±2.47)μ mol/L,P=0.000]also increased in oxidatively stressed + methionine group.Compared with oxidatively stressed group,the concentrations of homocysteine[(3.76 ± 0.22)vs.(1.67 ± 0.13)μmol/L,P =0.000],cysteine[(199.80 ± 8.75)vs.(82.64±15.88)μmol/L,P=0.000],and glutathione[(1.50 ±0.14)vs.(1.00 ±0.11)μ mol/L,P=0.011)]significantly increased in methionine group,and the concentrations of homocysteine[(3.84 ± 0.34)vs.(1.67±0.13)μmol/L,P=0.000],cysteine[(200.66±8.60)vs.(82.64±15.88)μmol/L,P=0.000]and glutathione[(1.40 ± 0.30)vs.(1.00 ± 0.11)μmol/L,P =0.028]significantly increased in oxidatively stressed + methionine groups.Compared with the control group,the contents of serine[(12.41 ± 1.51)vs.(24.00 ±2.54)mg/L,P =0.000],glutamate[(33.31 ±0.17)vs.(43.10 ±0.52)mg/L,P =0.000]and glycine[(6.23 ± 0.18)vs.(24.66 ± 10.87)mg/L,P =0.000]significantly decreased,while taurine [(7.99 ±0.16)vs.(6.17 ±0.15)mg/L,P =0.000]increased significantly in oxidatively stressed group.Compared with the oxidatively stressed group,the concentrations of serine[(16.98 ± 0.39)vs.(12.41 ± 1.51)mg/L,P=0.006)]and glutamate[(35.44 ±0.82)vs.(33.31 ±0.17)mg/L,P =0.002]in methionine group significantly increased,while taurine[(3.77 ±0.16)vs.(7.99 ±0.16)mg/L,P =0.000]significantly decreased in methionine group.Compared with the methionine group,the contents of serine[(12.59 ± 0.66)vs.(16.98±0.39)mg/L,P=0.008],glutamate[(30.87±0.60)vs.(35.44±0.82)mg/L,P=0.000]significantly decreased while taurine[(4.37 ± 0.12)vs.(3.77 ± 0.16)mg/L,P =0.001]in oxidatively stressed + methionine group significantly increased.Conclusion Oxidative stress can somehow promote homocysteine production in methionine loading BRL rat hepatocytes,but it is not the main effects.
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