CAJ | 학술논문

目的研究异烟肼致小鼠肝损伤进程中核因子相关因子-2 (Nrf-2)及超氧化物歧化酶(SOD) mRNA的表达情况.方法昆明小鼠64只,通过随机数字表中的简单随机分组法将小鼠分为8组,实验组予以异烟肼90 mg· kg-1·d-1(按成人用药量换算得到)连续灌胃1d、3d、5d、7d、2周、3周和4周,每个时间点小鼠8只.对照组给予等容积蒸馏水灌胃2周.全自动生化检测仪检测血清ALT、AST水平;比色法检测肝组织中SOD活力和丙二醛含量;应用赛绿博实时荧光定量PCR法检测肝组织中Nrf-2、铜-锌超氧化物歧化酶(Cu-ZnSOD)、锰超氧化物歧化酶(MnSOD) mRNA表达情况.多组间比较方差齐性者采用单因素方差分析,SNK法分析组间差异;方差不齐的组间比较采用多个独立样本的秩和检验.结果随着用药时间的延长,2周、3周、4周组小鼠血清ALT高于对照组(F=13.425,P=0.000);4周组血清AST高于对照组(F=3.498,P=0.004).总SOD活力在5d、7d、2周时分别为(272.63±22.43)、(276.14±20.90)、(271.00±27.43) U/mg,显著低于对照组的(317.95±19.32) U/mg(F=3.769,P=0.002);用药2周后Cu-ZnSOD活力为(242.53±21.48) U/mg,低于对照组(281.92±24.52)U/mg,但4周组活力为(277.11±24.52) U/mg,显著高于2周组(F=3.499,P=0.004);丙二醛含量在7d、2周、4周时分别为(0.94±0.28)、(0.91±0.23)、(0.80±0.15) nmol/mg,显著高于对照组的(0.54±0.09) nmol/mg(F=3.816,P=0.002).实时荧光定量PCR显示,用药4周后Nrf-2 mRNA表达比值增至对照组的6.24倍(F=2.987,P=0.014);Cu-ZnSOD mRNA在3周、4周分别增至2.29和3.99倍(F=8.261,P＜0.01);MnSOD在3周时表达增至4.85倍(F=4.026,P=0.002).结论在异烟肼致肝损伤发生、发展过程中,可上调SOD基因的表达,抵抗氧化损伤,同时可见Nrf-2基因的上调.
목적 연구이연정치소서간손상진정중핵인자상관인자-2 (Nrf-2)급초양화물기화매(SOD) mRNA적표체정황.방법 곤명소서64지,통과수궤수자표중적간단수궤분조법장소서분위8조,실험조여이이연정90 mg· kg-1·d-1(안성인용약량환산득도)련속관위1d、3d、5d、7d、2주、3주화4주,매개시간점소서8지.대조조급여등용적증류수관위2주.전자동생화검측의검측혈청ALT、AST수평;비색법검측간조직중SOD활력화병이철함량;응용새록박실시형광정량PCR법검측간조직중Nrf-2、동-자초양화물기화매(Cu-ZnSOD)、맹초양화물기화매(MnSOD) mRNA표체정황.다조간비교방차제성자채용단인소방차분석,SNK법분석조간차이;방차불제적조간비교채용다개독립양본적질화검험.결과 수착용약시간적연장,2주、3주、4주조소서혈청ALT고우대조조(F=13.425,P=0.000);4주조혈청AST고우대조조(F=3.498,P=0.004).총SOD활력재5d、7d、2주시분별위(272.63±22.43)、(276.14±20.90)、(271.00±27.43) U/mg,현저저우대조조적(317.95±19.32) U/mg(F=3.769,P=0.002);용약2주후Cu-ZnSOD활력위(242.53±21.48) U/mg,저우대조조(281.92±24.52)U/mg,단4주조활력위(277.11±24.52) U/mg,현저고우2주조(F=3.499,P=0.004);병이철함량재7d、2주、4주시분별위(0.94±0.28)、(0.91±0.23)、(0.80±0.15) nmol/mg,현저고우대조조적(0.54±0.09) nmol/mg(F=3.816,P=0.002).실시형광정량PCR현시,용약4주후Nrf-2 mRNA표체비치증지대조조적6.24배(F=2.987,P=0.014);Cu-ZnSOD mRNA재3주、4주분별증지2.29화3.99배(F=8.261,P＜0.01);MnSOD재3주시표체증지4.85배(F=4.026,P=0.002).결론 재이연정치간손상발생、발전과정중,가상조SOD기인적표체,저항양화손상,동시가견Nrf-2기인적상조.
Objective To observe the expressions of nuclear factor erythroid 2-related factor-2 (Nrf2) and superoxide dismutase (SOD) mRNA in mouse liver injury induced by isonicotinic acid hydrazide (isoniazid; INH).Methods A total of 64 Kunming mice were randomly divided into 8 groups using random number table:7 experimental groups and one control group.The mice in experimental groups were given isoniazid by intragastric administration at 90 mg/kg body weight for 1 d,3 d,5 d,7 d,2 weeks,3 weeks and 4 weeks with 8 mice at each time point.The mice in control group were given distilled water by intragastric administration for 2 weeks.Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured using an automatic biochemical analyzer,and liver SOD activity and methane dicarboxylic aldehyde (MDA) content were detected by colorimetric methods.SYBR green real-time polymerase chain reaction was used to test the expressions of Nrf2,Cu-ZnSOD and MnSOD mRNA in the liver.Comparison among multiple groups with homogeneity of variance was analyzed using analysis of variance and the difference between groups was analyzed by SNK method.Comparison between groups with heterogeneity of variance was analyzed using independent-samples rank sum test.Results The ALT levels at 2 weeks,3 weeks and 4 weeks in INH groups were significantly higher than that in control group (F=13.425,P=0.000); the AST level at 4 weeks in INH group was significantly higher than that in control group (F=3.498,P=0.004).The total activities of SOD at 5 d,7 d and 2 weeks in INH groups were (272.63±22.43),(276.14±20.90) and (271.00±27.43) U/mg,respectively,which were significantly lower compared with control group ([317.95 ± 19.32] U/mg,F=3.769,P=0.002).The activity of Cu-ZnSOD at 2 weeks in INH group ([242.53±21.48] U/mg) was significantly lower than that in control group ([281.92±24.52] U/mg),while it was significantly higher than that at4 weeks ([277.11±24.52] U/mg,F=3.499,P=0.004).TheMDAlevelsat7 d,2 weeks and 4 weeks were (0.94±0.28),(0.91±0.23) and (0.80±0.15) nmol/mg,which were significantly higher than that in control group ([0.54± 0.09] nmol/mg,F=3.816,P=0.002).Compared with control group,the Nrf-2 mRNA expression ratio in INH group had 6.42-fold increase at 4 weeks (F=2.987,P=0.014); the Cu-ZnSOD mRNA expression ratios at 3 weeks and 4 weeks were increased to 2.29 and 3.99 folds,respectively (F=8.261,P＜0.01) ; the MnSOD mRNA expression ratio was upregulated to 4.85 fold at 3 weeks (F=4.026,P=0.002).Conclusion During the development of liver injury induced by INH,the SOD gene expression is up-regulated to resist oxidative damage.Meanwhile,Nrf-2 gene is also up-regulated.