發(fā)布時(shí)間：2018-05-03 15:33

  本文選題：急性高脂血癥 + 大鼠�。� 參考：《河北醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:隨著人們生活方式和飲食結(jié)構(gòu)的改變,高脂血癥(Hyperglipemia,HLP)發(fā)病率逐年升高。高脂血癥能增加動(dòng)脈壁細(xì)胞內(nèi)自由基釋放系統(tǒng)的活性,損傷動(dòng)脈壁抗氧化機(jī)能,導(dǎo)致血管內(nèi)皮損傷,引起內(nèi)皮形態(tài)及功能異常,終至動(dòng)脈粥樣硬化(atherosclerosis,AS)形成。超氧化物歧化酶(Superoxide dismutase,SOD)和丙二醛(Malondialdehyde,MDA)是反應(yīng)氧化-抗氧化平衡的經(jīng)典指標(biāo)。核因子E2相關(guān)因子2(Nuclear factor E-2-related 2,Nrf2)是抗氧化應(yīng)激的核心轉(zhuǎn)錄因子。早期預(yù)防和治療高脂血癥,可以延緩和減少動(dòng)脈粥樣硬化性疾病的發(fā)生,提高人群的生活質(zhì)量,降低致殘致死率。本實(shí)驗(yàn)通過對急性高脂血癥大鼠預(yù)防性應(yīng)用姜黃素,觀察急性高脂狀態(tài)對內(nèi)皮形態(tài)和功能的影響以及氧化應(yīng)激指標(biāo)的變化,探討姜黃素保護(hù)內(nèi)皮、調(diào)節(jié)氧化應(yīng)激狀態(tài)的作用和機(jī)制。方法:1大鼠分組及預(yù)防性用藥:健康成年雄性Sprague-Dawley大鼠40只,按體重隨機(jī)分為正常對照組(NC組,n=8)、模型組(n=8只)、低劑量姜黃素組(125mg/kg.d姜黃素化合物組,LC組,n=8)、高劑量姜黃素組(250mg/kg.d姜黃素化合物組,HC組,n=8)、阿托伐他汀組(他汀組,n=8)。NC組、模型組給予2.5ml/kg玉米油灌胃,LC組、HC組、他汀組分別給予125mg/kg.d、250mg/kg.d姜黃素化合物及2.1mg/kg.d阿托伐他汀(均為2.5ml/kg)每日灌胃1次,連續(xù)10d。2制備急性高脂血癥大鼠模型:第10d灌胃1h后,模型組、LC組、HC組、他汀組均給予75%蛋黃乳液25ml/kg腹腔注射制造急性高脂血癥模型,NC組給予生理鹽水25ml/kg腹腔注射。3一般情況觀察:試驗(yàn)期間每天觀察各組大鼠的活動(dòng)情況、精神狀態(tài),記錄體重變化。4制備標(biāo)本及檢測指標(biāo):腹腔注射蛋黃乳液或生理鹽水24h后取血檢測總膽固醇(TC),甘油三酯(TG)、高密度脂蛋白膽固醇(HDL-C)、低密度脂蛋白膽固醇(LDL-C)、谷丙轉(zhuǎn)氨酶(ALT)、谷草轉(zhuǎn)氨酶(AST)、葡萄糖(GLU)、氧化低密度脂蛋白膽固醇(ox LDL)、SOD活力、一氧化氮(NO)。然后麻醉大鼠取腹主動(dòng)脈,部分采用戊二醛低溫速凍切片,用于HE染色觀察形態(tài)學(xué)變化,免疫組化法測定腹主動(dòng)脈中Nrf2表達(dá)水平;部分-80℃冷藏用于測定腹主動(dòng)脈MDA含量。5統(tǒng)計(jì)學(xué)分析符合正態(tài)分布計(jì)量資料以x±s表示,采用SPSS13.0軟件進(jìn)行統(tǒng)計(jì)學(xué)分析,多組間數(shù)據(jù)比較用單因素方差分析,兩兩比較用LSD檢驗(yàn),P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:1一般情況各組大鼠一般狀況良好,體重穩(wěn)定增長,反應(yīng)靈敏。各組大鼠體重差別無統(tǒng)計(jì)學(xué)意義(P0.05)。2注射蛋黃乳液24h后大鼠血脂升高,表現(xiàn)為TC、TG、LDL-C升高,HDL-C降低,認(rèn)為急性高脂血癥模型成立。3血脂水平3.1模型組大鼠血脂與NC組比較結(jié)果與NC組比較,模型組大鼠血清TC、TG、LDL-C水平升高,血清HDL-C水平降低(P0.05)。3.2藥物干預(yù)組大鼠血脂與模型組比較結(jié)果HC組、他汀組血清TC、TG水平均較模型組降低(P0.05);LC組血清TC、TG較模型組有降低趨勢,但無統(tǒng)計(jì)學(xué)差異(P0.05)。LC組、HC組、他汀組血清LDL-C水平較模型組均有降低趨勢,但無統(tǒng)計(jì)學(xué)差異(P0.05)。HC組血清HDL-C與模型組比較,無統(tǒng)計(jì)學(xué)差異(P0.05)。LC組、他汀組血清HDL-C則較模型組降低(P0.05)。3.3藥物干預(yù)組間血脂比較結(jié)果HC組、LC組、他汀組TC、TG、LDL-C水平無統(tǒng)計(jì)學(xué)差異(P0.05)。HC組血清HDL-C較LC組、他汀組升高(P0.05),LC組、他汀組血清HDL-C無統(tǒng)計(jì)學(xué)差異(P0.05)。4肝功能、葡萄糖水平各組大鼠肝功能指標(biāo)、血糖無統(tǒng)計(jì)學(xué)差異(P0.05)。5腹主動(dòng)脈HE染色結(jié)果NC組大鼠腹主動(dòng)脈內(nèi)膜光滑、完整,無隆起,平滑肌及彈力纖維排列規(guī)則,未見平滑肌細(xì)胞增生,外膜未見異常;模型組大鼠腹主動(dòng)脈內(nèi)膜不連續(xù),局部隆起,部分脫落,中膜平滑肌細(xì)胞及彈力纖維排列紊亂,外膜未見異常。LC組大鼠腹主動(dòng)脈內(nèi)膜連續(xù),局部隆起,彈力纖維、平滑肌細(xì)胞排列不規(guī)則;HC組大鼠腹主動(dòng)脈內(nèi)膜完整,無隆起,彈力纖維及平滑肌細(xì)胞排列稍不規(guī)則;他汀組大鼠腹主動(dòng)脈內(nèi)膜完整,局部可見隆起,彈力纖維及平滑肌細(xì)胞排列不規(guī)則。6免疫組織化學(xué)結(jié)果模型組腹主動(dòng)脈的Nrf2陽性細(xì)胞數(shù)較NC組增加(+44%,P0.05)。與模型組相比,LC組(+69%,P0.05)、HC組(+147%,P0.05)和他汀組(+51%,P0.05)的Nrf2陽性細(xì)胞數(shù)均顯著增加。與他汀組比較,LC組(+12%,P0.05)Nrf2陽性細(xì)胞數(shù)顯著增加,HC組(+64%,P0.05)Nrf2陽性細(xì)胞數(shù)顯著增加,且HC組(+46%,P0.05)Nrf2陽性細(xì)胞數(shù)多于LC組。7 SOD、MDA結(jié)果血清SOD活力:模型組SOD活力較NC組減弱(P0.05)。LC組、HC組、他汀組SOD活力與模型組相比增強(qiáng)(P0.05)。HC組、他汀組SOD活力與LC組比較增強(qiáng)(P0.05),HC組、他汀組SOD活力無統(tǒng)計(jì)學(xué)差異。腹主動(dòng)脈MDA含量:模型組MDA含量高于NC組(P0.05);LC組、HC組、他汀組的MDA含量低于模型組(P0.05)。HC組的MDA含量低于LC組和他汀組(P0.05),LC組、他汀組的MDA含量無統(tǒng)計(jì)學(xué)差異(P0.05)。8血清NO模型組NO含量低于NC組(P0.05)。LC組、HC組、他汀組NO含量與模型組相比均有顯著增加(P0.05)。HC組NO含量高于LC組和他汀組(P0.05),LC組和他汀組NO含量無統(tǒng)計(jì)學(xué)差異(P0.05)。9 ox LDL結(jié)果模型組血清ox LDL含量高于NC組(P0.05)。HC組、他汀組的ox LDL含量與模型組相比降低(P0.05),LC組ox LDL含量與模型組相比無統(tǒng)計(jì)學(xué)差異(P0.05)。HC組和他汀組ox LDL含量均低于LC組(P0.05),HC組和他汀組ox LDL含量無統(tǒng)計(jì)學(xué)差異(P0.05)。結(jié)論:1應(yīng)用75%蛋黃乳液腹腔注射成功制備急性高脂大鼠模型。急性高脂大鼠模型出現(xiàn)了腹主動(dòng)脈內(nèi)皮損傷,包括形態(tài)學(xué)改變(內(nèi)膜不光滑)和功能損傷(NO含量降低);急性高脂大鼠抗氧化能力減弱表現(xiàn)為脂質(zhì)過氧化產(chǎn)物ox LDL、MDA增加,SOD活力減弱。2低劑量姜黃素(125mg/kg.d姜黃素化合物)、高劑量姜黃素(250mg/kg.d姜黃素化合物)及阿托伐他汀均可降低急性高脂血癥模型大鼠血脂,250mg/kg.d降脂效果優(yōu)于125mg/kg.d。姜黃素和阿托伐他汀可能通過上調(diào)腹主動(dòng)脈Nrf2表達(dá),增強(qiáng)動(dòng)脈抗氧化應(yīng)激能力,減輕腹主動(dòng)脈病理損傷。
[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.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R589.2

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