兰州大学机构库 >学院待认领
蛋氨酸负荷对去卵巢大鼠肝脏的影响及叶酸、N-乙酰半胱氨酸的干预作用
Alternative TitleThe effect of methionine overload on liver in ovariectomized rats and interventive effect of folic and N-acetylcysteine
王顺霞
Thesis Advisor汤旭磊
2010-05-25
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword肝损伤 去卵巢 蛋氨酸 高同型半胱氨酸血症 氧化应激 硝基酪氨酸 雌激素 叶酸 N-乙酰半胱氨酸
Abstract目的 观察高同型半胱氨酸血症(HHcy)对去卵巢大鼠肝脏的影响及雌激素、叶酸和N-乙酰半胱氨酸对HHcy所致肝损伤的保护作用,并探讨其机制。方法 将48只4月龄雌性SD大鼠按体重随机均分为6组:假手术组(Sham组)、去卵巢组(OVX组)、蛋氨酸负荷组(OVX + 蛋氨酸1 g. kg-1. d-1,Met组)、17-β雌二醇替代治疗组(OVX + Met + 17-β雌二醇0.1 mg.kg -1.d -1, ERT组)、叶酸组(OVX + Met + 叶酸20 mg. kg-1. d-1, FA组)和N-乙酰半胱氨酸组(OVX + Met + N-乙酰半胱氨酸100 mg. kg-1. d-1,NAC组)。去卵巢术后1周开始蛋氨酸灌胃、药物干预,用药9周后处死全部大鼠,留取标本,测定血浆总同型半胱氨酸浓度 (tHcy)、血清丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST),HE染色观察肝脏病理学变化,肝组织匀浆测定总抗氧化力(TAC)、丙二醛 (MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)水平、谷胱甘肽还原酶(GR)和还原型谷胱甘肽(GSH)含量,采用免疫组织化学染色和蛋白质印迹法检测肝组织硝化蛋白。结果 1. 与Sham组比较,OVX和Met组血浆tHcy水平明显升高 ( p < 0.01 ) ;与OVX组比,Met组血浆tHcy水平显著升高(p < 0.01);ERT、FA和NAC组血浆tHcy水平较Met组明显下降(p< 0.01),但仍显著高于Sham组水平( p < 0.01 )2. 与Sham组相比,OVX和Met组血浆ALT、AST水平显著升高(p < 0.01, p < 0.05);与OVX组比较,Met组血浆ALT、AST水平显著升高(p < 0.01);ERT、FA和NAC组血浆ALT、AST水平较Met组明显下降(p< 0.01),但仍高于Sham组(p < 0.01, p < 0.05);NAC组ALT、AST水平较ERT组下降(p < 0.05)。3. 肝组织形态学改变:Sham组肝小叶结构清晰;OVX组肝细胞及肝小叶结构改变,存在轻度脂肪变性;Met组肝正常组织结构被破坏,肝细胞广泛脂肪变性;ERT 、FA和NAC组与Met组比较,肝小叶结构及肝细胞脂肪变性均有不同程度的改善。4. 相较于Sham组,OVX和Met组肝组织匀浆中MDA水平显著升高(p< 0.01),TAC、SOD、GSH、GR和GSH-PX水平明显降低(p< 0.01);与OVX组比较,Met组MDA水平显著升高(p< 0.01),TAC、SOD、GSH、GR和GSH-PX水平明显降低(p < 0.01, p < 0.05);ERT、FA和NAC组与Met组比,MDA水平明显下降(p< 0.01),TAC、SOD、GSH、GR和GSH-PX水平显著升高(p < 0.01, p < 0.05)。5. 肝组织硝基酪氨酸(NT)的表达:OVX和Met组较Sham组的表达明显增加(p < 0.01),Met组高于OVX组(p < 0.01);ERT、FA和NAC组NT的表达水平较Met组明显下降(p< 0.01),但仍显著高于Sham组水平( p < 0.01 )。结论 1. 蛋氨酸负荷引起的高同型半胱氨酸血症诱发了去卵巢大鼠的肝损伤,其机制可能与高同型半胱氨酸血症诱导肝脏氧化损伤有关。 2. 17-β-雌二醇、叶酸和N-乙酰半胱氨酸可不同程度地改善HHcy所致去卵巢大鼠的肝损伤,其机制可能与改善大鼠同型半胱氨酸代谢和氧化应激状态等因素有关。
Other AbstractObjective To investigate the effect of Hyperhomocysteinema over the liver in ovariectomized rats and the positive action of estrogen,folic acid and N-acetylcysteine toward HHcy’s effect and to explore the mechanism. Methods 48 SD rats of 4 months were randomly divided into 6 groups, namely group of sham-operated ( Sham group ),ovariectomy ( OVX group ),methionine ( OVX + L-methionine 1 g.kg-1.d-1,Met group ),estrogen replacement therapy ( OVX + L-methionine + 17-β estradiol 0.1 mg.kg -1.d -1,ERT group ),folic acid ( OVX + L-methionine + folic 20 mg.kg-1.d-1,FA group ) and N-acetylcysteine( OVX+ L-methionine+ N-acetylcysteine 100 mg.kg-1.d-1,NAC group ).1 week after been ovariectomized,all rats were intervened with drugs and gastric perfusion.Then,9 weeks later,all the rats were sacrificed for checking the level of total homocysteine( tHcy ),alanine aminotransferase ( ALT ),aspartate aminotransferase ( AST ).The pathological changes were observed after haematoxylin-eosin ( HE ) staining in liver.Total antioxidative capacity ( TAC ),malondialdehyde ( MDA ),superoxide dismutase ( SOD ) and glutathion peroxidase ( GSH-PX ) and concentration of glutathione reductase ( GR ) and reduced glutathione hormone ( GSH ) in their liver homogenates.Besides,the nitrotyrosine of liver were tested through method of immunohistochemical staining and western blot. Results 1.Compared with Sham group,serum tHcy concentrations were increased significantly in OVX and Met groups( p < 0.01).Compared with OVX group,serum tHcy concentrations were increased distinctly in Met group ( p < 0.01).ERT,FA and NAC groups presented a decrease of serum tHcy compared to Met group ( p < 0.01 ),which were still significantly higher than Sham group ( p < 0.01).2.Serum ALT and AST levels had significant rise in OVX and Met group (p < 0.01, p < 0.05) compared to Sham group.Compared with OVX group,serum ALT and AST levels were increased distinctly in Met group ( p < 0.01).ERT,FA and NAC groups presented a significant decrease of serum ALT and AST levels compared to Met group ( p < 0.01 ),which were still higher than Sham group (p < 0.01, p < 0.05).Moreover,the ALT and AST levels in NAC group were lower than ERT group(p < 0.01).3.Morphological observation in liver tissue: The structure of hepatic lobules is clear in Sham group.In OVX group,the liver cells and hepatic lobules have changed,and there is mild steatosis.The normal liver structure was destroyed and extensive fatty degeneration of ...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223660
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
王顺霞. 蛋氨酸负荷对去卵巢大鼠肝脏的影响及叶酸、N-乙酰半胱氨酸的干预作用[D]. 兰州. 兰州大学,2010.
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