D-柠檬烯改善大鼠酒精性肝损伤实验的研究

发布时间：2018-06-16 18:30

本文选题：D-柠檬烯 + ALD　；参考：《青岛大学》2014年硕士论文

【摘要】：目的观察D-柠檬烯(D-limonene)对酒精性肝损伤大鼠抗氧化活性及对肝组织核因子κB(NF-κB)、环氧合酶2(Cyclooxygenase-2,COX-2)蛋白表达的影响。方法雄性Wistar大鼠共70只,按体重随机分为7组。A组为空白对照组;B组为酒精模型组,每日给予50%乙醇灌胃,前2周8 m L/(kg?d),后4周递增为12m L/(kg?d),继续灌胃4周;C、D、E组分别为D-柠檬烯低、中、高剂量组,每日分别给予D-柠檬烯100、200、400 mg/kg灌胃;F组为D-柠檬烯单独高剂量组,每日给予D-柠檬烯400mg/kg灌胃;G组为甘利欣对照组,每日给予甘利欣200mg/kg。C、D、E、G组酒精剂量同模型组,F组蒸馏水剂量同空白对照组。每组10只,实验进行6周。末次灌胃禁食12 h后,麻醉,取血,分离血清,留取肝组织。HE染色观察肝脏组织的病理学改变;检测大鼠血浆和肝匀浆中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)水平;免疫组化法检测肝组织中NF-κB、COX-2表达情况,计算其阳性表达率。结果HE染色结果显示,酒精模型组大鼠肝组织呈大泡性脂肪变性,并有大量炎性细胞的浸润;D-柠檬烯各剂量组与酒精模型组相比,脂肪变性呈不同程度减轻,炎性细胞减少。与空白对照组相比,酒精模型组血浆SOD、血浆和肝匀浆GSH-Px活力均降低,血浆和肝匀浆MDA含量明显升高(P0.05);D-柠檬烯中、高剂量组血浆SOD以及各剂量组血浆和肝匀浆GSH-Px活力均高于酒精模型组,血浆和肝匀浆MDA含量则低于酒精模型组(P0.05)。酒精模型组与空白对照组相比,NF-κB、COX-2表达明显增多,而D-柠檬烯各剂量组NF-κB、COX-2表达均低于酒精模型组,差异有显著性(P0.05)。与空白对照组相比,F组大鼠各指标无明显差异(P0.05)。结论D-柠檬烯能够增强酒精性肝损伤大鼠体内抗氧化酶的活性,减少脂质过氧化物的产生,减少肝组织NF-κB、COX-2蛋白表达,从而改善因酒精暴露引起的肝损伤。
[Abstract]:Aim to observe the effects of D-limonene on the antioxidant activity and the expression of nuclear factor 魏 B (NF- 魏 B) -NF- 魏 B (cyclooxygenase-2cyclooxygenase-2COX-2) protein in rats with alcoholic liver injury. Methods A total of 70 male Wistar rats were randomly divided into 7 groups according to their body weight. Group A was a blank control group and group B was treated with alcohol model group. 50% ethanol was administered daily by gavage. In the first 2 weeks, 8 mL 路L / L / kg ~ (-1) d ~ (-1) was added to 12 mL 路L ~ (-1) 路kg ~ (-1) d ~ (-1) at the last 4 weeks. After 4 weeks of continuous intragastric administration, the low, middle and high doses of D- limonene were given to the group C ~ (2 +) and D ~ (100200400) mg/kg daily respectively. The group F was treated with high dose of Dlimonene alone, respectively, after 4 weeks of oral administration of D-limonene. Group G was treated with D-limonene 400mg/kg intragastrically as control group, and the alcohol dosage of group C was 200 mg / kg / d. The dose of distilled water in group F was the same as that in group C (P < 0.05), and the dose of distilled water in group F was the same as that in group C (P < 0.05). The experiment was conducted for 6 weeks with 10 rats in each group. After the last gastric fasting for 12 hours, the liver was anesthetized, blood samples were taken, serum was isolated, liver tissue was stained with HE to observe the pathological changes of liver tissue. The levels of superoxide dismutase (SOD), malondialdehyde (MDAA) and glutathione peroxidase (GSH-Px) in plasma and liver homogenate of rats were detected, and the expression of COX-2 in liver tissue was detected by immunohistochemical method, and the positive rate of COX-2 was calculated. Results HE staining showed that the liver tissue of the alcohol model group showed vesicular steatosis, and there were a large number of inflammatory cells infiltrated in D- limonene groups. Compared with the alcohol model group, the steatosis was reduced in varying degrees and the inflammatory cells were decreased in each dose group. Compared with control group, plasma SOD, GSH-Px activity in plasma and liver homogenate in alcohol model group decreased, MDA content in plasma and liver homogenate increased significantly in D- limonene. The activity of GSH-Px in plasma and liver homogenate in high dose group was higher than that in alcohol model group, and MDA content in plasma and liver homogenate was lower than that in alcohol model group. Compared with the control group, the expression of COX-2 in the alcohol model group was significantly higher than that in the control group, while the expression of COX-2 in the D- limonene group was lower than that in the alcohol model group (P 0.05). Compared with the blank control group, there was no significant difference in each index between the F group and the blank control group (P 0.05). Conclusion D- limonene can enhance the activity of antioxidant enzymes, reduce the production of lipid peroxides and decrease the expression of NF- 魏 B _ (1) COX-2 protein in the liver tissue of rats with alcoholic liver injury, thereby improving the liver injury induced by alcohol exposure.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575

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