姜黄素对急性高脂血症大鼠氧化应激及腹主动脉Nrf2表达的影响

发布时间：2018-05-03 15:33

本文选题：急性高脂血症 + 大鼠　；参考：《河北医科大学》2015年硕士论文

【摘要】：目的:随着人们生活方式和饮食结构的改变,高脂血症(Hyperglipemia,HLP)发病率逐年升高。高脂血症能增加动脉壁细胞内自由基释放系统的活性,损伤动脉壁抗氧化机能,导致血管内皮损伤,引起内皮形态及功能异常,终至动脉粥样硬化(atherosclerosis,AS)形成。超氧化物歧化酶(Superoxide dismutase,SOD)和丙二醛(Malondialdehyde,MDA)是反应氧化-抗氧化平衡的经典指标。核因子E2相关因子2(Nuclear factor E-2-related 2,Nrf2)是抗氧化应激的核心转录因子。早期预防和治疗高脂血症,可以延缓和减少动脉粥样硬化性疾病的发生,提高人群的生活质量,降低致残致死率。本实验通过对急性高脂血症大鼠预防性应用姜黄素,观察急性高脂状态对内皮形态和功能的影响以及氧化应激指标的变化,探讨姜黄素保护内皮、调节氧化应激状态的作用和机制。方法:1大鼠分组及预防性用药:健康成年雄性Sprague-Dawley大鼠40只,按体重随机分为正常对照组(NC组,n=8)、模型组(n=8只)、低剂量姜黄素组(125mg/kg.d姜黄素化合物组,LC组,n=8)、高剂量姜黄素组(250mg/kg.d姜黄素化合物组,HC组,n=8)、阿托伐他汀组(他汀组,n=8)。NC组、模型组给予2.5ml/kg玉米油灌胃,LC组、HC组、他汀组分别给予125mg/kg.d、250mg/kg.d姜黄素化合物及2.1mg/kg.d阿托伐他汀(均为2.5ml/kg)每日灌胃1次,连续10d。2制备急性高脂血症大鼠模型:第10d灌胃1h后,模型组、LC组、HC组、他汀组均给予75%蛋黄乳液25ml/kg腹腔注射制造急性高脂血症模型,NC组给予生理盐水25ml/kg腹腔注射。3一般情况观察:试验期间每天观察各组大鼠的活动情况、精神状态,记录体重变化。4制备标本及检测指标:腹腔注射蛋黄乳液或生理盐水24h后取血检测总胆固醇(TC),甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、葡萄糖(GLU)、氧化低密度脂蛋白胆固醇(ox LDL)、SOD活力、一氧化氮(NO)。然后麻醉大鼠取腹主动脉,部分采用戊二醛低温速冻切片,用于HE染色观察形态学变化,免疫组化法测定腹主动脉中Nrf2表达水平;部分-80℃冷藏用于测定腹主动脉MDA含量。5统计学分析符合正态分布计量资料以x±s表示,采用SPSS13.0软件进行统计学分析,多组间数据比较用单因素方差分析,两两比较用LSD检验,P0.05为差异有统计学意义。结果:1一般情况各组大鼠一般状况良好,体重稳定增长,反应灵敏。各组大鼠体重差别无统计学意义(P0.05)。2注射蛋黄乳液24h后大鼠血脂升高,表现为TC、TG、LDL-C升高,HDL-C降低,认为急性高脂血症模型成立。3血脂水平3.1模型组大鼠血脂与NC组比较结果与NC组比较,模型组大鼠血清TC、TG、LDL-C水平升高,血清HDL-C水平降低(P0.05)。3.2药物干预组大鼠血脂与模型组比较结果HC组、他汀组血清TC、TG水平均较模型组降低(P0.05);LC组血清TC、TG较模型组有降低趋势,但无统计学差异(P0.05)。LC组、HC组、他汀组血清LDL-C水平较模型组均有降低趋势,但无统计学差异(P0.05)。HC组血清HDL-C与模型组比较,无统计学差异(P0.05)。LC组、他汀组血清HDL-C则较模型组降低(P0.05)。3.3药物干预组间血脂比较结果HC组、LC组、他汀组TC、TG、LDL-C水平无统计学差异(P0.05)。HC组血清HDL-C较LC组、他汀组升高(P0.05),LC组、他汀组血清HDL-C无统计学差异(P0.05)。4肝功能、葡萄糖水平各组大鼠肝功能指标、血糖无统计学差异(P0.05)。5腹主动脉HE染色结果NC组大鼠腹主动脉内膜光滑、完整,无隆起,平滑肌及弹力纤维排列规则,未见平滑肌细胞增生,外膜未见异常;模型组大鼠腹主动脉内膜不连续,局部隆起,部分脱落,中膜平滑肌细胞及弹力纤维排列紊乱,外膜未见异常。LC组大鼠腹主动脉内膜连续,局部隆起,弹力纤维、平滑肌细胞排列不规则;HC组大鼠腹主动脉内膜完整,无隆起,弹力纤维及平滑肌细胞排列稍不规则;他汀组大鼠腹主动脉内膜完整,局部可见隆起,弹力纤维及平滑肌细胞排列不规则。6免疫组织化学结果模型组腹主动脉的Nrf2阳性细胞数较NC组增加(+44%,P0.05)。与模型组相比,LC组(+69%,P0.05)、HC组(+147%,P0.05)和他汀组(+51%,P0.05)的Nrf2阳性细胞数均显著增加。与他汀组比较,LC组(+12%,P0.05)Nrf2阳性细胞数显著增加,HC组(+64%,P0.05)Nrf2阳性细胞数显著增加,且HC组(+46%,P0.05)Nrf2阳性细胞数多于LC组。7 SOD、MDA结果血清SOD活力:模型组SOD活力较NC组减弱(P0.05)。LC组、HC组、他汀组SOD活力与模型组相比增强(P0.05)。HC组、他汀组SOD活力与LC组比较增强(P0.05),HC组、他汀组SOD活力无统计学差异。腹主动脉MDA含量:模型组MDA含量高于NC组(P0.05);LC组、HC组、他汀组的MDA含量低于模型组(P0.05)。HC组的MDA含量低于LC组和他汀组(P0.05),LC组、他汀组的MDA含量无统计学差异(P0.05)。8血清NO模型组NO含量低于NC组(P0.05)。LC组、HC组、他汀组NO含量与模型组相比均有显著增加(P0.05)。HC组NO含量高于LC组和他汀组(P0.05),LC组和他汀组NO含量无统计学差异(P0.05)。9 ox LDL结果模型组血清ox LDL含量高于NC组(P0.05)。HC组、他汀组的ox LDL含量与模型组相比降低(P0.05),LC组ox LDL含量与模型组相比无统计学差异(P0.05)。HC组和他汀组ox LDL含量均低于LC组(P0.05),HC组和他汀组ox LDL含量无统计学差异(P0.05)。结论:1应用75%蛋黄乳液腹腔注射成功制备急性高脂大鼠模型。急性高脂大鼠模型出现了腹主动脉内皮损伤,包括形态学改变(内膜不光滑)和功能损伤(NO含量降低);急性高脂大鼠抗氧化能力减弱表现为脂质过氧化产物ox LDL、MDA增加,SOD活力减弱。2低剂量姜黄素(125mg/kg.d姜黄素化合物)、高剂量姜黄素(250mg/kg.d姜黄素化合物)及阿托伐他汀均可降低急性高脂血症模型大鼠血脂,250mg/kg.d降脂效果优于125mg/kg.d。姜黄素和阿托伐他汀可能通过上调腹主动脉Nrf2表达,增强动脉抗氧化应激能力,减轻腹主动脉病理损伤。
[Abstract]:Objective: with the change of lifestyle and dietary structure, the incidence of Hyperglipemia (HLP) increases year by year. Hyperlipidemia can increase the activity of free radical release system in the arterial wall cells, damage the antioxidant function of the arterial wall, cause vascular endothelial damage, induce endothelial morphology and function abnormality, and end to atherosclerosis (ather Osclerosis, AS) formation. Superoxide dismutase (Superoxide dismutase, SOD) and malondialdehyde (Malondialdehyde, MDA) are the classic indicators of the reaction oxidation - antioxidant balance. Nuclear factor E2 related factor 2 (Nuclear factor E-2-related 2, Nrf2) is a nuclear cardiac transcription factor of antioxidant stress. Early prevention and treatment of hyperlipidemia can be delayed and reduced. In this experiment, the effect of curcumin on the morphology and function of endothelium and the changes of oxidative stress were observed through the preventive application of curcumin on acute hyperlipidemia rats, and the protection of curcumin to protect the endothelium and regulate the oxidative stress. Methods: the function and mechanism of state. Methods: 1 rats group and preventive medicine: 40 healthy adult male Sprague-Dawley rats were randomly divided into normal control group (group NC, n=8), model group (n=8), low dose curcumin group (125mg/kg.d curcumin compound, LC group, n=8), high dose curcumin group, HC group, n=8. In the Atorvastatin group (statins group, n=8) group.NC, the model group was given 2.5ml/kg corn oil for gastric perfusion, LC, HC, and statins were given 125mg/kg.d, 250mg/kg.d curcumin compound and 2.1mg/kg.d atrovastatin (2.5ml/kg) were given 1 times a day, and the rat model of acute hyperlipidemia was prepared by 10d.2. Group, statins group were given 75% Egg Yolk Emulsion 25ml/kg intraperitoneal injection of acute hyperlipidemia model, group NC given saline 25ml/kg intraperitoneal injection of.3 general condition observation: during the test, observe the activity of rats in each group, mental state, record body weight change.4 preparation standard and detection index: intraperitoneal injection of yolk lotion or physiological salt After 24h, blood was taken to detect total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), glutamic pyruvidase (ALT), glutamine transaminase (AST), glucose (GLU), oxidized low density lipoprotein cholesterol (ox LDL), SOD vitality, nitric oxide (NO). Then the abdominal aorta of rats was anesthetized and partially used. The low temperature frozen section of glutaraldehyde was used to observe the morphological changes in HE staining. The level of Nrf2 expression in abdominal aorta was measured by immunohistochemical method. The MDA content of abdominal aorta was measured at -80 degrees centigrade, and the statistical analysis of.5 in the abdominal aorta was in accordance with the normal distribution measurement data with x + s, and SPSS13.0 soft parts were used for statistical analysis, and the data of multiple groups were compared with single data. Factor variance analysis, 22 compared with LSD test, P0.05 has statistical significance. Results: 1 general conditions of the general condition of rats in general condition, stable weight growth, reaction sensitive. No statistical significance (P0.05).2 injection of Yolk Emulsion 24h after 24h rats blood lipid increase, the expression of TC, TG, LDL-C increase, HDL-C lower, think that Acute hyperlipidemia model set up.3 blood lipid level 3.1 model group rats blood lipid and group NC compared with the NC group, the model group rats serum TC, TG, LDL-C level increased, serum HDL-C level decreased (P0.05).3.2 drug intervention group rats blood lipid and model group of HC group, statin group serum TC, TG levels were lower than the model group. The serum TC and TG were lower than the model group, but there was no statistical difference (P0.05).LC group, HC group, and statin group had a lower serum LDL-C level than the model group, but there was no statistical difference (P0.05), there was no statistical difference between the serum HDL-C and the model group (P0.05).LC group (P0.05), and the statin group serum HDL-C was lower than the model group. There was no statistical difference between group HC, group LC, statin group, TC, TG and LDL-C (P0.05), the serum HDL-C of.HC group was higher than that of LC group, the statins group increased (P0.05), LC group, and the statins group had no statistical difference in liver function, and the liver function index of the rats in each group of glucose was not statistically different. In group NC, the abdominal aorta intima was smooth, complete, no bulge, smooth muscle and elastic fiber arrangement, no smooth muscle cell proliferation, no abnormal outer membrane, the model group of abdominal aorta intima discontinuous, partial uplift, partial shedding, middle membrane smooth muscle cells and elastic fiber arrangement disorder, outer membrane no abnormal group of.LC rats abdominal initiative In group HC rats, the abdominal aorta intima was intact, no bulge, elastic fibers and smooth muscle cells were arranged slightly irregular. The abdominal aorta intima of the rats in the statin group was intact, the elastic fibers and smooth muscle cells arranged irregular.6 immunohistochemical results. The number of Nrf2 positive cells in the abdominal aorta of the model group was higher than that in the NC group (+44%, P0.05). Compared with the model group, the number of positive cells in the LC group (+69%, P0.05), HC group (+147%, P0.05) and statins group (+51%, P0.05) increased significantly. The number of Nrf2 positive cells in group HC (+46%, P0.05) was more than that of LC group.7 SOD, and MDA results in serum SOD activity: SOD activity in the model group was weaker than that of NC group (P0.05). The content of MDA in the model group was higher than that of the NC group (P0.05), and the content of MDA in the LC group, the HC group and the statin group was lower than that of the model group (P0.05). The content of the MDA content was lower than that of the LC group and the statin group (P0.05), and the content of the statin group was lower than that of the NC group. The content of the statin group was lower than that of the group NC. The content of NO in group.HC was higher than that of group LC and statin group (P0.05), and there was no significant difference in NO content between LC and statins group (P0.05).9 ox LDL result group was higher than that of the group, and the content of statin group was lower than that of the model group, and there was no statistical difference between the model group and the model group. The content of ox LDL in.HC group and statin group was lower than that in group LC (P0.05), and there was no statistical difference between HC and statin group ox LDL (P0.05). Conclusion: 1 the acute high fat rat model was successfully prepared by intraperitoneal injection of 75% egg yolk emulsion. The decrease of O content, the decrease of antioxidant capacity in acute high fat rats was ox LDL, MDA and.2 low dose curcumin (125mg/kg.d curcumin). The high dose curcumin (curcumin compound) and atorvastatin could reduce the blood lipid of the acute hyperlipidemia model rats and reduce the blood lipid in 250mg/kg.d. The effect of 125mg/kg.d. curcumin and atorvastatin may be enhanced by increasing the expression of Nrf2 in the abdominal aorta, enhancing the anti oxidative stress of the arteries and alleviating the pathological injury of the abdominal aorta.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R589.2

【参考文献】