本文選題：染料木素 + 動脈粥樣硬化　； 參考：《南華大學(xué)》2011年碩士論文

【摘要】：目的:利用高脂飲食和低劑量、低毒性有機(jī)磷農(nóng)藥敵百蟲腹腔注射建立SD大鼠動脈粥樣硬化模型,觀察染料木素抗大鼠動脈粥樣硬化作用,并檢測血脂、丙二醛含量等生化指標(biāo),初步探討其作用機(jī)制,為染料木素的合理用藥提供實(shí)驗(yàn)和理論依據(jù)。 方法: 健康SD大鼠56只分為七組,其中普通飼料喂養(yǎng)(普食)即溶媒對照組(1 ml·kg~(-1)·day~(-1))、高劑量染料木素(genistein ,Gen)對照組(5mg·kg~(-1)·day~(-1))和陽性藥物辛伐他汀對照組(5mg·kg~(-1)·day~(-1))三組,高膽固醇高脂飼料喂養(yǎng)(簡稱高脂)加敵百蟲即模型對照組[10%的二甲亞砜(DMSO),1ml·kg~(-1)·day~(-1)]、低劑量Gen對照組(0.5mg·kg~(-1)·day~(-1))、高劑量Gen對照組(5mg·kg~(-1)·day~(-1))和陽性藥辛伐他汀對照組(5 mg·kg~(-1)·day~(-1))四組。 2周為大鼠稱一次體重,根據(jù)體重調(diào)整給藥劑量。給藥后繼續(xù)按原飼料方案喂養(yǎng)8周。在實(shí)驗(yàn)給藥前和給藥后第4、8周分別斷尾采血,使用怡成全血葡萄糖測定儀檢測大鼠血糖(Glu),用德國羅氏診斷有限公司試劑盒檢測大鼠血清總膽固醇(TC)、甘油三酯(TG)、高密度脂蛋膽固醇(HDL-C)、低密度脂蛋白膽固醇(LDL-C)、C反應(yīng)蛋白(CRP)含量;實(shí)驗(yàn)第4、8周末用分光光度法檢測血清對氧磷酶(PON1)活性,用Griess試劑顯色法測定大鼠血清總一氧化氮(NO),硫代巴比妥法檢測大鼠血清中丙二醛(MDA)濃度;第8周末處死所有動物,采用羥肟酸鐵比色法測定大鼠全血乙酰膽堿酯酶(Acetylcholine esterase, AchE)活性,摘取大鼠肝臟及主動脈弓,肉眼和蘇木素-伊紅(HE)染色后觀察各組大鼠主動脈弓及肝臟病理形態(tài)學(xué)變化;同時摘取心臟分離其周圍組織后稱重,計(jì)算其臟器指數(shù)。使用免疫組化SP法檢測主動脈血管壁內(nèi)皮型一氧化氮合酶(eNOS)的蛋白表達(dá)情況。 結(jié)果: 1實(shí)驗(yàn)大鼠一般狀況 普食組大鼠毛色光澤,活動自如,活潑,體重增長較快;高脂喂養(yǎng)加敵百蟲的四組大鼠隨著實(shí)驗(yàn)進(jìn)程,毛色松散暗淡,活動減少,其中給予低、高劑量Gen和辛伐他汀的三組大鼠精神狀態(tài)明顯好轉(zhuǎn),體重都呈現(xiàn)先降低在增長,且實(shí)驗(yàn)期間體重變化不大。 2血脂含量變化 第4、8周末,普食喂養(yǎng)的三組中大鼠血清血脂含量、HDL-C/TC與AI值均無顯著差異(P0.05);高脂喂養(yǎng)加敵百蟲的四組大鼠血清TC、LDL-C、HDL-C含量與AI值均較普食喂養(yǎng)的三組明顯升高(P0.05或P0.01或P0.001),而HDL-C/TC值明顯降低(P0.05或P0.01)。各組大鼠血清TG含量無顯著差異(P0.05)。Gen能明顯降低TC、LDL-C,使AI降低,HDL-C/TC比值升高。 3大鼠GLU含量變化 Gen各劑量組和辛伐他汀均能明顯降低動脈粥樣硬化大鼠血糖水平(P 0.05或P0.001)。 4血清CRP含量變化 高劑量Gen和辛伐他汀能明顯降低動脈粥樣硬化大鼠血清炎癥因子CRP的含量(P0.05)。 5血清MDA含量變化 高劑量Gen和辛伐他汀能明顯降低血清MDA水平。 6血清NO含量變化 高脂喂養(yǎng)加敵百蟲的四組大鼠血清NO含量較普食喂養(yǎng)的三組明顯升高(P0.05或P0.01);高脂喂養(yǎng)加敵百蟲的四組大鼠中,給予Gen和辛伐他汀組與高脂喂養(yǎng)加敵百蟲模型對照組大鼠血清NO含量均無顯著差異(P0.05)。 7血清PON1含量變化 Gen提高As大鼠血清PON1活性(P0.05或P0.01),存在量效關(guān)系。 8組織形態(tài)觀察 肝臟與主動脈弓形態(tài)學(xué)觀察,Gen的各劑量組和辛伐他汀均能在一定程度上對血管壁具有保護(hù)作用。 9 Gen對主動脈eNOS蛋白表達(dá)的影響 免疫組化結(jié)果:Gen各劑量組和辛伐他汀均能在一定程度上增加eNOS表達(dá)。 結(jié)論: 1.成功建立敵百蟲加速的SD大鼠As模型。 2.Gen能明顯抑制敵百蟲加速的SD大鼠As的發(fā)生,改善其肝與主動脈弓形態(tài)變化。其可能的機(jī)制與逆轉(zhuǎn)血脂障礙,通過抑制過氧化脂質(zhì)作用抗氧化,通過降低炎癥因子CRP抗炎,并參NO體系、eNOS與PON1酶的調(diào)節(jié)有關(guān)。
[Abstract]:Objective: to establish the atherosclerotic model of SD rats by intraperitoneal injection of high fat diet and low toxic organophosphorus pesticide didiworm intraperitoneally, observe the effect of genistein on atherosclerosis and detect the biochemical indexes of blood lipid and malondialdehyde content, and explore the mechanism of its action to provide experiment and reason for the rational use of genistein. On the basis.
Method:
56 healthy SD rats were divided into seven groups, of which common feed was fed (1 ml. Kg~ (-1). Day~ (-1)), high dose genistein (genistein, Gen) control group (5mg kg~ (-1) day~) and positive drug simvastatin control group (-1). The two methylene sulfoxide (DMSO), 1ml / kg~ (-1) / day~ (-1)) of the model control group, the low dose Gen control group (0.5mg kg~ (-1) day~ (day~)), the high dose control group (kg~) and the positive drug simvastatin control group (1ml) four groups.
The rats were weighed once for 2 weeks, and the dosage was adjusted according to the weight of the rats. After the drug was given for 8 weeks, the blood was collected before and after the 4,8 week, and the blood glucose (Glu) was detected by the Yicheng whole blood glucose analyzer, and the serum total cholesterol (TC) was detected by the German Roche Diagnostics Ltd kit. The content of triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and C reactive protein (CRP), the activity of serum to oxyphosphatase (PON1) was detected by spectrophotometric method at the end of 4,8 at the end of the experiment, and the serum total nitric oxide (NO) was determined by the chromogenic reagent of Griess reagent, and malondialdehyde (MDA) in the serum of rats was detected by thiobarbituric method. All the animals were killed at the end of the eighth week. The activity of Acetylcholine esterase (AchE) in the whole blood of rats was measured by ferrous hydroxamic acid colorimetry. The liver and aortic arch of rats were extracted, the naked eye and hematoxylin eosin (HE) were stained to observe the pathological changes of the aortic arch and liver in each group, and the circumferential separation of the heart was also taken. The organs were weighed and the Viscera index was calculated. The expression of endothelial nitric oxide synthase (eNOS) in the aortic wall was detected by immunohistochemical SP.
Result:
1 general condition of experimental rats
The four groups of rats in the four groups of rats with high fat feeding and stinworm were given low, high dose and simvastatin. The three groups of rats with high dose and simvastatin showed a significant improvement in the mental state of the rats, and the body weight decreased first and the body weight changed during the experiment. Not much.
Changes of blood lipid content in 2
At the end of the 4,8 weekend, there was no significant difference in serum lipid content in the three groups of the rats fed by universal diet (P0.05). The levels of TC, LDL-C, HDL-C and AI in the serum of four groups of rats fed with high fat feeding and the HDL-C/TC were significantly higher than those of the three groups feeding on the universal diet (P0.05 or P0.01 or P0.001). There was no significant difference in TG content (P0.05)..Gen significantly decreased TC and LDL-C, reduced AI and increased HDL-C/TC ratio.
Changes of GLU content in 3 rats
Gen dose and simvastatin could significantly reduce the blood glucose level of atherosclerosis rats (P 0.05 or P0.001).
Change of serum CRP content in 4
High dose of Gen and simvastatin could significantly reduce the content of serum inflammatory factor CRP in atherosclerotic rats (P0.05).
Change of serum MDA content in 5
High dose of Gen and simvastatin significantly decreased serum MDA level.
Change of serum NO content in 6
The serum NO content of four groups of rats fed with high fat feeding and a total of the three groups were significantly increased (P0.05 or P0.01), and there was no significant difference in the content of NO in the serum of the high fat feeding and the four groups of the simvastatin group and the high fat feeding and the rat model control group (P0.05).
Change of serum PON1 content in 7
Gen increased the activity of serum PON1 (P0.05 or P0.01) in As rats, and there was a dose effect relationship.
8 observation of tissue morphology
Morphologic observation of liver and aortic arch showed that all doses of Gen and simvastatin could protect the vascular wall to some extent.
The effect of 9 Gen on the expression of eNOS protein in the aorta
Immunohistochemical results: Gen dose and simvastatin could increase eNOS expression to some extent.
Conclusion:
1. the As model of SD rats was successfully established.
2.Gen can obviously inhibit the occurrence of As in SD rats accelerated by DTM and improve the morphological changes of the hepatic and aortic arch. Its possible mechanism is related to reversing lipid disorders, inhibiting the antioxidant activity of lipid peroxide, reducing inflammation factor CRP by reducing inflammation and regulating the regulation of eNOS and the regulation of PON1 enzyme in the NO system.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R363

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