【摘要】：背景:梗阻性黃疸是指肝內(nèi)膽管或肝外膽管等膽道系統(tǒng)發(fā)生機(jī)械性梗阻時(shí),膽紅素因膽汁向腸道的排泄受阻而返流入血所引起的黃疸。膽管受到阻塞時(shí)可有血清膽紅素及總膽汁酸明顯增高,而血清堿性磷酸酶(ALP)、谷氨酰轉(zhuǎn)移酶(GGT)、谷丙轉(zhuǎn)氨酶(ALT)等指標(biāo)也可有顯著增高。由于膽紅素的親脂性特點(diǎn),其易與脂質(zhì)含量多的組織結(jié)合而使組織細(xì)胞受損傷,而中樞神經(jīng)系統(tǒng)又是機(jī)體除脂肪組織外脂質(zhì)含量最豐富的器官,因此,膽紅素對(duì)中樞神經(jīng)系統(tǒng)損傷是最大的。膽紅素濃度的升高可干擾正常腦細(xì)胞代謝,使細(xì)胞處于氧化應(yīng)激狀態(tài),產(chǎn)生過(guò)多的氧自由基;并下調(diào)大腦中閉合蛋白的濃度,導(dǎo)致血腦屏障的破壞。于此同時(shí)大腦中caveolin-1(與腦血管屏障的損壞有關(guān))和β-catenin的表達(dá)也可增加,以減輕膽紅素對(duì)血腦屏障的損害。此外,膽紅素對(duì)神經(jīng)系統(tǒng)的損傷還產(chǎn)生大量的小分子物質(zhì),產(chǎn)生的氧自由基及脂質(zhì)過(guò)氧化物可消耗大量的抗氧化酶,使體內(nèi)沉積大量的高活性分子,氧化系統(tǒng)及抗氧化系統(tǒng)失去平衡,進(jìn)而對(duì)組織及細(xì)胞產(chǎn)生氧化應(yīng)激損傷。氧化應(yīng)激損傷使生物膜損傷更重,當(dāng)生物膜嚴(yán)重受損時(shí),大量鈣離子流入細(xì)胞內(nèi),可引起細(xì)胞內(nèi)Ca2+超載,造成細(xì)胞功能障礙或喪失。在此基礎(chǔ)上,小膠質(zhì)細(xì)胞被激活,誘發(fā)炎癥反應(yīng),最終導(dǎo)致神經(jīng)營(yíng)養(yǎng)因子的缺乏,甚至引起神經(jīng)細(xì)胞的進(jìn)行性變性及死亡。Chroni E等研究發(fā)現(xiàn)了梗阻性黃疸可以導(dǎo)致大鼠大腦中直接和間接氧化應(yīng)激標(biāo)記物的增加或降低,證實(shí)在梗阻性黃疸早期和晚期時(shí)的氧化應(yīng)激狀態(tài)與黃疸有關(guān)聯(lián),指出氧化應(yīng)激是膽紅素介導(dǎo)的腦病的一個(gè)重要機(jī)制。最近關(guān)于氧化應(yīng)激反應(yīng)的研究表明,Keap1-Nrf2-ARE抗氧化通路在機(jī)體抗氧化損傷的過(guò)程中起著重要的作用,它能維持細(xì)胞內(nèi)氧化還原平衡,使細(xì)胞免受氧化性谷氨酸毒性和過(guò)氧化氫誘導(dǎo)的凋亡。NMDAR(N-methyl-D-aspartate receptor,N-甲基-D-天冬氨酸受體)是離子型谷氨酸受體一個(gè)亞型,主要分布于中樞神經(jīng)系統(tǒng)的突觸后膜。NMDAR主要參與興奮性突觸傳遞,具有配體電壓雙重門控的特性,對(duì)鈣離子有較高的通透性。NMDAR的過(guò)度激活可引起興奮性神經(jīng)毒性,并通過(guò)一系列的反應(yīng),最終導(dǎo)致神經(jīng)細(xì)胞死亡。而氧自由基也可以促進(jìn)神經(jīng)元和神經(jīng)膠質(zhì)細(xì)胞釋放谷氨酸,同時(shí)又可以抑制谷氨酸的再攝取,谷氨酸激活NMDA受體通過(guò)Ca2+活化磷脂酶A又可產(chǎn)生氧自由基從而形成惡性循環(huán)。地卓西平馬來(lái)酸鹽(dizocilpinemaleate,MK-801)是一種非競(jìng)爭(zhēng)性NMDA受體拮抗劑,它易于透過(guò)血腦屏障,對(duì)NMDAR通道進(jìn)行變構(gòu)調(diào)節(jié),可有效阻滯谷氨酸(glutamic acid)與突觸后膜上的受體結(jié)合,阻止NMDAR偶聯(lián)的Ca2+通道激活,減少Ca2+內(nèi)流,進(jìn)而使NMDA受體的作用減弱,降低谷氨酸的毒性作用。大量研究結(jié)果證實(shí)MK-801對(duì)中毒、缺血缺氧、肝性腦病等多種情況下的神經(jīng)元損傷具有保護(hù)作用。.白藜蘆醇(resveratrol,Res)是在葡萄和紅酒中大量存在的一種黃酮類多酚化合物,大量的體內(nèi)和體外研究發(fā)現(xiàn)它有廣泛的生物學(xué)效應(yīng)和藥學(xué)活性作用,如抵御活性自由基對(duì)機(jī)體細(xì)胞DNA的損傷并且防止細(xì)胞膜發(fā)生脂質(zhì)過(guò)氧化反應(yīng)等,并且已在心血管疾病、糖尿病、腎臟疾病等證實(shí),發(fā)現(xiàn)白藜蘆醇所產(chǎn)生的藥理作用至少有部分是通過(guò)誘導(dǎo)HO-1的表達(dá)而發(fā)揮抗氧化應(yīng)激損傷而進(jìn)一步產(chǎn)生細(xì)胞保護(hù)作用的,但目前尚無(wú)在梗阻性黃疸發(fā)生時(shí)中樞神經(jīng)系統(tǒng)中的應(yīng)用研究。本實(shí)驗(yàn)首先建立梗阻性黃疸大鼠模型,再通過(guò)觀察MK-801對(duì)梗阻性黃疸引起的大腦氧化應(yīng)激反應(yīng)中丙二醛(MDA),總抗氧化能力(T-AOC),超氧化物歧化酶(SOD),過(guò)氧化氫酶(CAT),核轉(zhuǎn)錄因子E2相關(guān)因子2(transcription factor NF-E2-related factor 2,Nrf2)蛋白,Nfr2 m RNA,血紅素加氧酶-1(heineoxygenase-1,HO-1),NAD(P)H醌氧化還原酶1(NQO1,NAD(P)H quinone oxidoreductase 1)等指標(biāo)的影響,進(jìn)而深入探討NMDA受體阻滯劑對(duì)梗阻性黃疸所致大腦氧化應(yīng)激損傷是否具有保護(hù)作用及其具體機(jī)制。論文第三部分主要通過(guò)白藜蘆醇對(duì)梗阻性黃疸大鼠中樞神經(jīng)系統(tǒng)內(nèi)MDA,SOD及HO-1研究其在氧化應(yīng)激損傷的過(guò)程中的作用。第一部分MK-801對(duì)梗阻性黃疸大鼠中樞神經(jīng)抗氧化應(yīng)激系統(tǒng)的影響目的:本部分研究的目的在于成功建立梗阻性黃疸大鼠模型,并探討MK-801對(duì)梗阻性黃疸大鼠中樞神經(jīng)抗氧化應(yīng)激系統(tǒng)的影響。方法:20只大鼠隨機(jī)分為四組:I組(假手術(shù)組)、II組(對(duì)照組)、III組(低劑量處理組)和IV組(高劑量處理組),II,III,IV組為梗阻性黃疸模型組,III,IV組為MK-801組。第2天起III組腹腔注射MK-801,0.025mg/kg.d,IV組腹腔注射MK-801,0.25mg/kg.d;I組和II組以等容量生理鹽水同時(shí)注射。四組大鼠術(shù)后3d采尾部靜脈血以全自動(dòng)生化儀檢查血清總膽紅素和直接膽紅素濃度來(lái)確定模型制作是否成功,術(shù)后10天處死大鼠并取材(大腦皮質(zhì))按試劑盒要求行MDA及CAT、T-SOD、T-AOC含量或活性測(cè)定。結(jié)果:1假手術(shù)組實(shí)驗(yàn)鼠術(shù)后當(dāng)日即恢復(fù)飲食及自由活動(dòng),且活動(dòng)靈敏,小便淡黃。梗阻性黃疸模型組大鼠第2天開始出現(xiàn)尾尖及耳部等皮膚較薄的部位黃染,并逐漸出現(xiàn)全身黃染,尿液深黃,精神萎靡、反應(yīng)遲緩。術(shù)后第三天,對(duì)照組、MK-801低劑量組、MK-801高劑量組大鼠大腦組織中直接膽紅素的濃度和總膽汁酸含量(μmol/L)與假手術(shù)組相比明顯增高,且有統(tǒng)計(jì)學(xué)差異(P0.01),考慮梗阻性黃疸模型制作成功。2對(duì)照組、MK-801低劑量組、MK-801高劑量組丙二醛的含量與假手術(shù)組相比升高,且有統(tǒng)計(jì)學(xué)差異(P0.05)。應(yīng)用MK-801后丙二醛含量與對(duì)照組相比降低,且具有統(tǒng)計(jì)學(xué)意義,但MK-801高、低劑量組之間無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。3對(duì)照組氧化氫酶活性與假手術(shù)組相比降低,MK-801組過(guò)氧化氫酶活性比對(duì)照組明顯增高,且有統(tǒng)計(jì)學(xué)差異(P0.05)。MK-801低劑量組、MK-801高劑量組之間過(guò)氧化氫酶活性無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)4對(duì)照組總超氧化物歧化酶活性比假手術(shù)組降低,且有統(tǒng)計(jì)學(xué)意義,MK-801組總超氧化物岐化酶活性與對(duì)照組相比明顯增高,且有統(tǒng)計(jì)學(xué)差異(P0.05)。MK-801高劑量組總超氧化物岐化酶活性與低劑量組相比無(wú)統(tǒng)計(jì)學(xué)差異。5對(duì)照組、MK-801低劑量組、MK-801高劑量組總抗氧化能力與假手術(shù)組相比明顯增高,且有統(tǒng)計(jì)學(xué)意義(P0.05)。MK-801組比對(duì)照組總抗氧化能力升高,但高劑量組與對(duì)照組之間相比無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。結(jié)論1通過(guò)結(jié)扎實(shí)驗(yàn)動(dòng)物膽總管可以成功建立梗阻性黃疸動(dòng)物模型。2梗阻性黃疸時(shí)中樞神經(jīng)系統(tǒng)機(jī)體存在氧化應(yīng)激反應(yīng),機(jī)體通過(guò)增加過(guò)氧化氫酶等抗氧化應(yīng)激損傷酶類的表達(dá)使抗氧化應(yīng)激損傷能力增強(qiáng),這與國(guó)內(nèi)外目前研究一致。3 MK-801使梗阻性黃疸所致大鼠中樞神經(jīng)系統(tǒng)氧化應(yīng)激損傷的過(guò)程中脂質(zhì)過(guò)氧化程度降低,并且可以提高SOD的活性及機(jī)體總抗氧化應(yīng)激能力,高劑量的MK-801與低劑量的MK-801相比效果并未明顯增加,反而使T-SOD活性比低劑量組降低,具體機(jī)制需要進(jìn)一步研究。第二部分Keap1-Nrf2-ARE信號(hào)通路在梗阻性黃疸大鼠中樞神經(jīng)系統(tǒng)氧化應(yīng)激損傷中的作用探討目的:研究Keap1-Nrf2-ARE信號(hào)通路在梗阻性黃疸大鼠中樞神經(jīng)系統(tǒng)氧化應(yīng)激損傷中的作用及在機(jī)體氧化應(yīng)激反應(yīng)過(guò)程中NMDA受體阻滯劑對(duì)Keap1-Nrf2-ARE信號(hào)通路的影響方法:四組大鼠分別為:I組(假手術(shù)組)、II組(對(duì)照組)、III組(低劑量處理組)和IV組(高劑量處理組),II,III,IV組為梗阻性黃疸模型組,第2天起III組腹腔注射MK-801,0.025mg/kg.d,IV組腹腔注射MK-801,0.25mg/kg.d,連續(xù)注射10d;I組和II組以等容量生理鹽水同時(shí)注射。術(shù)后10天取材以Western bolt行HO-1,NQO1,Nrf2蛋白,以RT-PCR行Nrf2 m RNA測(cè)定。結(jié)果1與假手術(shù)組相比,OJ組Nrf2 m RNA表達(dá)均明顯升高,且均有統(tǒng)計(jì)學(xué)差異,P0.05。MK-801組與對(duì)照組相比Nrf2 m RNA表達(dá)升高且有統(tǒng)計(jì)學(xué)意義,P0.05。高劑量組與對(duì)照組相比Nrf2 m RNA表達(dá)降低,但無(wú)統(tǒng)計(jì)學(xué)差異,P0.05。2與假手術(shù)組相比,OJ組Nrf2蛋白表達(dá)均增高,且有統(tǒng)計(jì)學(xué)意義,P0.05;MK-801組與對(duì)照組相比Nrf2蛋白表達(dá)增高,且有統(tǒng)計(jì)學(xué)意義,P0.05。不同的是高劑量組與低劑量組相比Nrf2蛋白表達(dá)減低,而且具有統(tǒng)計(jì)學(xué)意義,P0.05。3 OJ組與假手術(shù)組相比HO-1蛋白表達(dá)增高,且有統(tǒng)計(jì)學(xué)意義,P0.05。MK-801組與對(duì)照組相比HO-1蛋白表達(dá)增高,且有統(tǒng)計(jì)學(xué)意義,P0.05。高低劑量MK-801組比低劑量組相比,HO-1表達(dá)水平降低,且具有統(tǒng)計(jì)學(xué)差異P0.05。4OJ組與假手術(shù)組相比NQO1蛋白表達(dá)增高,且有統(tǒng)計(jì)學(xué)意義,P0.05。MK-801組與對(duì)照組相比NQO1蛋白表達(dá)增高,且有統(tǒng)計(jì)學(xué)意義,P0.05。高低劑量MK-801組比低劑量組相比,NQO1表達(dá)水平降低,且具有統(tǒng)計(jì)學(xué)差異P0.05。結(jié)論1再一次驗(yàn)證梗阻性黃疸時(shí)機(jī)體存在氧化應(yīng)激反應(yīng)。2機(jī)體通過(guò)Keap1-Nrf2-ARE信號(hào)通路增強(qiáng)了HO-1和NQO1及Nrf2蛋白的表達(dá),增強(qiáng)了自身抗氧化應(yīng)激能力。3 NMDA受體阻滯劑MK-801通過(guò)Keap1-Nrf2-ARE信號(hào)通路上調(diào)了II相解毒酶HO-1和NQO-1的蛋白表達(dá),增加了機(jī)體抗氧化應(yīng)激反應(yīng)能力,從而發(fā)揮了它的神經(jīng)保護(hù)作用。雖然高、低劑量的MK-801均能達(dá)到此作用,但高劑量的MK-801反而使Nrf2 m RNA、Nrf2蛋白、HO-1和NQO1蛋白的表達(dá)較低劑量組降低,可能與MK-801毒性作用有關(guān)。第三部分不同水平白藜蘆醇對(duì)梗阻性黃疸大鼠中樞神經(jīng)系統(tǒng)氧化應(yīng)激目的:通過(guò)對(duì)研究梗阻性黃疸大鼠大腦皮質(zhì)中氧化應(yīng)激指標(biāo)表達(dá)水平來(lái)探討不同水平白藜蘆醇對(duì)梗阻性黃疸大鼠中樞神經(jīng)系統(tǒng)氧化應(yīng)激損傷的影響及其氧化應(yīng)激損傷中的保護(hù)作用和機(jī)制。方法:選擇32只6周齡的雄性SD大鼠作為實(shí)驗(yàn)對(duì)象,將其隨機(jī)分為四個(gè)組別,分籠飼養(yǎng),每組8只,A組大鼠進(jìn)行梗阻性黃疸假手術(shù)(只將膽總管游離出來(lái),不做結(jié)扎),B、C、D組大鼠均進(jìn)行手術(shù)制備梗阻性黃疸模型,術(shù)后每日C、D兩組大鼠分別給予不同劑量的白藜蘆醇溶液灌胃,A、B組則給予同等劑量生理鹽水灌胃。分別在手術(shù)后的第14d采集血液標(biāo)本行總膽紅素、直接膽紅素、谷丙轉(zhuǎn)氨酶、谷草轉(zhuǎn)氨酶測(cè)定,并采集大腦皮質(zhì)標(biāo)本,并對(duì)其中中丙二醛、超氧化物歧化酶活性、HO-1水平進(jìn)行檢測(cè),觀察不同組別的大鼠各項(xiàng)指標(biāo)之間的差異。結(jié)果:術(shù)后TBIL及DBIL指標(biāo)結(jié)果證明II、III、IV三組大鼠梗阻性黃疸模型制作成功。OJ組ALT、AST、MDA水平比假手術(shù)組升高,SOD、HO-1水平比假手術(shù)組降低。OJ組內(nèi)反應(yīng)肝功能損害指標(biāo)ALT,AST及反應(yīng)氧化應(yīng)激損傷指標(biāo)MDA的平均水平呈IIIIIIV的趨勢(shì),而氧化應(yīng)激保護(hù)性指標(biāo)SOD和HO-1平均值則呈IIIIIIV的趨勢(shì),且具有統(tǒng)計(jì)學(xué)意義。不同劑量的白藜蘆醇組之間,指標(biāo)相比也存在統(tǒng)計(jì)學(xué)差異(P0.05)。結(jié)論:白藜蘆醇對(duì)梗阻性大鼠TBIL及DBIL影響不大,但可以保護(hù)其肝功能,并且可以能促進(jìn)大腦皮質(zhì)細(xì)胞組織中的丙二醛含量降低、SOD和HO-1活性升高,發(fā)揮抗氧化應(yīng)激損傷的作用。
[Abstract]:BACKGROUND: Obstructive jaundice refers to jaundice caused by bilirubin reflux into the blood due to obstruction of excretion of bile from the intestinal tract when mechanical obstruction occurs in the intrahepatic or extrahepatic bile ducts. Alpha-aminotransferase (ALT) and other indicators can also be significantly increased. Because of the lipophilic characteristics of bilirubin, it is easy to combine with lipid-rich tissues and cause tissue and cell damage. The central nervous system is the most lipid-rich organ except adipose tissue. Therefore, bilirubin is the greatest damage to the central nervous system. The elevation of Caveolin-1 and beta-catenin in the brain can also increase the expression of caveolin-1 and beta-catenin in the brain to reduce the blood-brain barrier damage. In addition, bilirubin damage to the nervous system also produces a large number of small molecules, oxygen free radicals and lipid peroxides can consume a large number of antioxidant enzymes, so that a large number of highly active molecules deposited in the body, oxidative system and antioxidant system imbalance, and then produce oxidative stress damage to tissues and cells. Oxidative stress injury can aggravate the damage of biofilm. When the biofilm is severely damaged, a large amount of calcium ions flow into the cell, which can cause intracellular Ca2+ overload, resulting in cell dysfunction or loss. On this basis, microglia are activated, causing inflammation, eventually leading to the lack of neurotrophic factors, and even lead to the progress of nerve cells. Chroni E and other studies have found that obstructive jaundice can lead to the increase or decrease of direct and indirect oxidative stress markers in the brain of rats, confirming that oxidative stress in the early and late stages of obstructive jaundice is associated with jaundice, indicating that oxidative stress is an important mechanism of bilirubin-mediated encephalopathy. Studies on oxidative stress have shown that the Keap1-Nrf2-ARE antioxidant pathway plays an important role in the process of anti-oxidative damage in the body. It can maintain the balance of oxidation and reduction in cells, and protect cells from oxidative glutamate toxicity and hydrogen peroxide-induced apoptosis. NMDAR (N-methyl-D-aspartate receptor, N-methyl-D-aspartate receptor) receives NMDAR is a subtype of ionic glutamate receptor, mainly distributed in the postsynaptic membranes of the central nervous system. NMDAR is involved in excitatory synaptic transmission, has the characteristics of ligand voltage double-gated, and has a high permeability to calcium ions. Overactivation of NMDAR can cause excitatory neurotoxicity, and ultimately lead to a series of reactions. Neuronal cell death. Oxygen free radicals also promote the release of glutamate from neurons and glial cells, and inhibit glutamate reuptake. Glutamate-activated NMDA receptors produce oxygen free radicals through Ca2+ activated phospholipase A to form a vicious cycle. Dizocilpine maleate (MK-801) is a kind of dizocilpine maleate. Non-competitive NMDA receptor antagonists, which can easily regulate NMDAR channels through blood-brain barrier, can effectively block the binding of glutamic acid to receptors on postsynaptic membranes, prevent the activation of NMDAR-coupled Ca2+ channels, reduce Ca2+ influx, thereby weakening the role of NMDA receptors and reducing the toxicity of glutamate. Resveratrol (Res) is a kind of flavonoid polyphenols in grape and red wine. A large number of in vivo and in vitro studies have found that it has a wide range of biological effects and pharmacological activities, such as. Resveratrol has been shown to protect cells from DNA damage by reactive free radicals and lipid peroxidation. It has been demonstrated in cardiovascular diseases, diabetes mellitus and kidney diseases that the pharmacological effects of resveratrol are at least partially mediated by inducing the expression of HO-1 and exerting antioxidant stress damage to cells. In this study, we first established a rat model of obstructive jaundice, and then observed the effects of MK-801 on malondialdehyde (MDA), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), hydrogen peroxide (hydrogen peroxide) in oxidative stress of the brain induced by obstructive jaundice. Enzyme (CAT), transcription factor NF-E2-related factor 2 (Nrf2) protein, Nfr2 m RNA, heme oxygenase-1 (HO-1), NAD (P) H quinone oxidoreductase 1 (NQO1, NAD (P) and other indicators of the impact of NMDA receptor blockers on obstructive jaundice caused by large The third part of this paper mainly studies the effects of resveratrol on MDA, SOD and HO-1 in the central nervous system of rats with obstructive jaundice. Objective: To establish a successful model of obstructive jaundice in rats and investigate the effects of MK-801 on the central nervous system in rats with obstructive jaundice.Methods: Twenty rats were randomly divided into four groups: group I (sham operation group), group II (control group), group III (low dose treatment group) and group IV (high dose treatment group), group II, III, IV. Group II was injected intraperitoneally with MK-801,0.025mg/kg.d. Group IV was injected intraperitoneally with MK-801,0.25mg/kg.d. Group I and Group II were injected with normal saline of the same volume at the same time. The rats were sacrificed 10 days after operation and the contents or activities of MDA, CAT, T-SOD and T-AOC were determined according to the requirements of the kit. Results: 1. The rats in sham operation group resumed their diet and free movement on the day after operation, and their urine was light yellow. The rats in obstructive jaundice model group began to appear tail tips and tail tips on the second day. On the third day after operation, the concentration of direct bilirubin and total bile acid (micromol/L) in brain tissue of rats in control group, low dose MK-801 group and high dose MK-801 group were significantly higher than those in sham operation group (P The content of malondialdehyde in control group, MK-801 low-dose group and MK-801 high-dose group was significantly higher than that in sham-operation group (P 0.05). The content of malondialdehyde in MK-801 high-dose group was significantly lower than that in control group (P 0.05). The catalase activity of control group was lower than that of sham operation group, and the catalase activity of MK-801 group was significantly higher than that of control group (P 0.05). The catalase activity of low dose MK-801 group and high dose MK-801 group had no statistical difference (P 0.05). The total superoxide dismutase activity of control group was lower than that of sham operation group. The total superoxide dismutase activity of MK-801 group was significantly higher than that of the control group, and there was a statistical difference (P 0.05). The total superoxide dismutase activity of MK-801 high-dose group was not significantly different from that of low-dose group. The total antioxidant capacity of MK-801 group was higher than that of the control group, but there was no significant difference between the high-dose group and the control group (P 0.05). Conclusion 1 The animal model of obstructive jaundice can be successfully established by ligating the common bile duct of experimental animals. 2 There is oxidative stress in the central nervous system during obstructive jaundice. In response, the body enhances the ability of anti-oxidative stress injury by increasing the expression of catalase and other anti-oxidative stress injury enzymes, which is consistent with the current research at home and abroad. 3 MK-801 can reduce the degree of lipid peroxidation in the process of oxidative stress injury of the central nervous system induced by obstructive jaundice in rats, and can increase the activity of SOD and The total antioxidant stress ability of the body, the effect of high dose MK-801 and low dose MK-801 did not increase significantly, but decreased the activity of T-SOD compared with low dose group. The specific mechanism needs further study. Part II The role of Keap1-Nrf2-ARE signaling pathway in oxidative stress injury of central nervous system in obstructive jaundice rats AIM: To investigate the role of Keap 1-Nrf 2-ARE signaling pathway in oxidative stress injury of central nervous system in rats with obstructive jaundice and the effect of NMDA receptor blockers on Keap 1-Nrf 2-ARE signaling pathway during oxidative stress in vivo. Methods: Four groups of rats were divided into: group I (sham operation group), group II (control group), group III (low dose treatment group) and group IV. Group II, III and IV were treated with intraperitoneal injection of MK-801, 0.025mg/kg.d from day 2. Group IV was treated with intraperitoneal injection of MK-801, 0.25mg/kg.d for 10 consecutive days. Group I and II were injected with normal saline of equal volume at the same time. Results 1 Compared with the sham operation group, the expression of Nrf2 m RNA in OJ group was significantly higher, and there was statistical difference. Compared with the control group, the expression of Nrf2 m RNA in OJ group was significantly higher and statistically significant, P 0.05. The expression of Nrf2 protein in MK-801 group was significantly higher than that in the control group (P 0.05). The expression of HO-1 protein in high and low dose MK-801 group was lower than that in low dose MK-801 group. The expression of NQO1 protein in high and low dose MK-801 group was higher than that in sham operation group (P 0.05). The expression of NQO1 was significantly lower in MK-801 group than in low dose group (P 0.05). Conclusion1 Again, there was an oxidative stress response in the body during obstructive jaundice. 2 The expression of HO-1, NQO1 and Nrf2 proteins was enhanced by Keap1-Nrf2-ARE signaling pathway. 3 NMDA receptor blocker MK-801 regulates the expression of phase II detoxifying enzymes HO-1 and NQO-1 through the Keap1-Nrf2-ARE signaling pathway, increases the antioxidant stress response ability of the body, and thus exerts its neuroprotective effect. Although both high and low doses of MK-801 can achieve this effect, high doses of MK-801 can induce Nrf2 m RNA instead. The expression of Nrf2 protein, HO-1 and NQO1 protein decreased in the lower dose group, which may be related to the toxicity of MK-801. Part III Oxidative stress of central nervous system in rats with obstructive jaundice induced by resveratrol at different levels: To investigate the expression of oxidative stress in cerebral cortex of rats with obstructive jaundice. Methods: Thirty-two six-week-old male SD rats were randomly divided into four groups and fed in cages. Eight rats in each group were given sham operation for obstructive jaundice. Rats in group B, C and D were operated on to establish obstructive jaundice model. Rats in group C and D were separated daily after operation.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R575

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 仲偉鑒,印木泉,浦躍樸;煤煙致大鼠肺細(xì)胞氧化應(yīng)激損傷的機(jī)制探討[J];勞動(dòng)醫(yī)學(xué);2000年03期

2 葛圣蕾;;自由基氧化應(yīng)激損傷在老齡化神經(jīng)變性疾病中的作用[J];國(guó)外醫(yī)學(xué)(老年醫(yī)學(xué)分冊(cè));2002年06期

3 王云;常志文;;硫氧還蛋白-1:一個(gè)新的疾病氧化應(yīng)激損傷標(biāo)志物[J];心臟雜志;2011年03期

4 丁銀慧;孫競(jìng);錢元霞;李蕓子;高靜;陳峗;;晚期糖基化終產(chǎn)物誘導(dǎo)人主動(dòng)脈內(nèi)皮細(xì)胞氧化應(yīng)激損傷及線粒體功能紊亂[J];江蘇大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2014年03期

5 劉朝巍;張濤;楊卓;;氧化應(yīng)激損傷線粒體參與癲癇病理過(guò)程[J];中國(guó)病理生理雜志;2008年01期

6 谷峰;董振南;賈興旺;王玲;田亞平;;抗震救災(zāi)醫(yī)療隊(duì)員與受災(zāi)人員氧化應(yīng)激損傷指標(biāo)的比較研究[J];標(biāo)記免疫分析與臨床;2010年06期

7 陳文豪;陳浩;趙達(dá)君;喬慧蓮;易定華;;番茄紅素對(duì)血管內(nèi)皮細(xì)胞氧化應(yīng)激損傷的作用及機(jī)制研究[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2014年11期

8 黎涌;招偉賢;;圍術(shù)期氧化應(yīng)激損傷及跨膜信號(hào)轉(zhuǎn)導(dǎo)研究進(jìn)展[J];國(guó)際麻醉學(xué)與復(fù)蘇雜志;2006年04期

9 張春霞;胡利民;康立源;劉利萍;郭虹;;腦缺血氧化應(yīng)激損傷及中藥拮抗作用研究進(jìn)展[J];中華中醫(yī)藥學(xué)刊;2007年12期

10 張德迎;魏光輝;何大維;劉星;林濤;李旭良;;葉下珠植物提取物保護(hù)鹽酸氮芥對(duì)小鼠睪丸組織的氧化應(yīng)激損傷[J];生殖與避孕;2008年12期

相關(guān)會(huì)議論文 前10條

1 顏建英;;子癇前期與氧化應(yīng)激損傷[A];中華醫(yī)學(xué)會(huì)第三次全國(guó)妊娠期高血壓疾病學(xué)術(shù)研討會(huì)論文匯編[C];2011年

2 王詠波;王茜;于振乾;杜建玲;;窖蛋白-1在波動(dòng)性高糖誘導(dǎo)的內(nèi)皮細(xì)胞氧化應(yīng)激損傷中的作用及機(jī)制[A];中華醫(yī)學(xué)會(huì)第十次全國(guó)內(nèi)分泌學(xué)學(xué)術(shù)會(huì)議論文匯編[C];2011年

3 呂萍萍;范瑩;陳瑩瑩;朱立;沈岳良;;環(huán)加氧酶2抑制劑對(duì)抗心肌氧化應(yīng)激損傷中的作用及機(jī)制[A];浙江省生理科學(xué)會(huì)2006年學(xué)術(shù)年會(huì)論文匯編[C];2006年

4 胡凱騫;張龍澤;陳春英;孫寶云;邢更妹;趙保路;趙宇亮;聶廣軍;;功能化富勒烯衍生物效應(yīng)探索:延長(zhǎng)線蟲壽命和保護(hù)氧化應(yīng)激損傷的分子機(jī)理研究[A];第十一次中國(guó)生物物理學(xué)術(shù)大會(huì)暨第九屆全國(guó)會(huì)員代表大會(huì)摘要集[C];2009年

5 韓婧;李學(xué)軍;;姜黃素通過(guò)誘導(dǎo)人內(nèi)皮細(xì)胞產(chǎn)生自噬抗氧化應(yīng)激損傷[A];全國(guó)第十二屆生化與分子藥理學(xué)學(xué)術(shù)會(huì)議論文集[C];2011年

6 方朝暉;鮑陶陶;章小平;;2型糖尿病中醫(yī)辨證與氧化應(yīng)激損傷相關(guān)性的研究[A];第九次全國(guó)中醫(yī)糖尿病學(xué)術(shù)大會(huì)論文匯編[C];2006年

7 張銘;陳懷生;楊大春;曾智;余敏;;同型半胱氨酸對(duì)血管內(nèi)皮細(xì)胞的氧化應(yīng)激損傷及對(duì)IL-8分泌的影響[A];全國(guó)第九屆心臟學(xué)會(huì)第十二屆心功能學(xué)會(huì)《心臟雜志》編委會(huì)聯(lián)合學(xué)術(shù)會(huì)議論文集[C];2005年

8 王艷霞;劉儀;張偉;韓東寧;張?jiān)词?;血管緊張素轉(zhuǎn)化酶2(ACE2)對(duì)大鼠腎氧化應(yīng)激損傷的保護(hù)作用及其機(jī)制[A];中國(guó)生理學(xué)會(huì)消化內(nèi)分泌生殖代謝生理專業(yè)委員會(huì)2011年消化內(nèi)分泌生殖學(xué)術(shù)會(huì)議論文摘要匯編[C];2011年

9 張婷;王凡;朱俊東;易龍;付鈺潔;糜漫天;;染料木黃酮對(duì)人血管內(nèi)皮細(xì)胞氧化應(yīng)激損傷的保護(hù)作用[A];重慶市營(yíng)養(yǎng)學(xué)會(huì)學(xué)術(shù)會(huì)議論文匯編[C];2010年

10 齊華林;王俊;江薇;嚴(yán)海東;;瘦素在誘導(dǎo)血管內(nèi)皮細(xì)胞氧化應(yīng)激損傷中的作用[A];2007年浙滬兩地腎臟病學(xué)術(shù)年會(huì)資料匯編[C];2007年

相關(guān)重要報(bào)紙文章 前3條

1 張中橋;復(fù)方丹參注射液對(duì)EPC氧化應(yīng)激損傷有保護(hù)作用[N];中國(guó)中醫(yī)藥報(bào);2007年

2 記者 衣曉峰;我國(guó)首設(shè)中醫(yī)神志病學(xué)博士后科研工作站[N];中國(guó)中醫(yī)藥報(bào);2014年

3 張中橋;復(fù)方丹參注射液對(duì)內(nèi)皮前體細(xì)胞氧化應(yīng)激損傷有保護(hù)作用[N];中國(guó)醫(yī)藥報(bào);2006年

相關(guān)博士學(xué)位論文 前9條

1 劉靜;氧化應(yīng)激損傷在赭曲霉毒素A誘導(dǎo)細(xì)胞周期阻滯中的作用及其可能機(jī)制的研究[D];河北醫(yī)科大學(xué);2012年

2 易龍;黃酮類植物化學(xué)物抑制內(nèi)皮細(xì)胞氧化應(yīng)激損傷的結(jié)構(gòu)—效應(yīng)關(guān)系研究[D];第三軍醫(yī)大學(xué);2010年

3 張立超;抗氧化劑在梗阻性黃疸大鼠中樞神經(jīng)系統(tǒng)氧化應(yīng)激損傷中的應(yīng)用研究[D];河北醫(yī)科大學(xué);2015年

4 符曉華;含二氟亞甲基異黃酮類化合物的合成及抗血管內(nèi)皮氧化應(yīng)激損傷活性測(cè)定[D];中南大學(xué);2006年

5 葉俊生;左卡尼汀在腎小管上皮細(xì)胞氧化應(yīng)激損傷中的保護(hù)作用研究[D];南方醫(yī)科大學(xué);2010年

6 張昱;PJ-34對(duì)心肌缺血再灌注損傷的保護(hù)作用及其抗氧化應(yīng)激損傷的機(jī)制研究[D];浙江大學(xué);2011年

7 陸志明;高游離脂肪酸誘導(dǎo)血管局部氧化應(yīng)激損傷的分子機(jī)制及中藥干預(yù)研究[D];四川大學(xué);2006年

8 王瑞敏;低劑量Genistein誘導(dǎo)eNOS/Nrf2-ARE通路對(duì)缺血后神經(jīng)元氧化應(yīng)激損傷和空間學(xué)習(xí)記憶缺陷的保護(hù)作用[D];河北醫(yī)科大學(xué);2013年

9 付志紅;UCP2在女性生殖中作用的研究[D];第一軍醫(yī)大學(xué);2005年

相關(guān)碩士學(xué)位論文 前10條

1 王莉;7-二氟亞甲基-5，，4’-二甲烷氧基異黃酮對(duì)過(guò)氧化氫誘導(dǎo)的血管內(nèi)皮氧化應(yīng)激損傷的保護(hù)作用研究[D];南華大學(xué);2006年

2 鄒玉凌;綠原酸對(duì)氧化應(yīng)激損傷人視網(wǎng)膜色素上皮細(xì)胞保護(hù)作用的分子機(jī)制[D];南昌大學(xué);2014年

3 鄭媛媛;Nrf2在大鼠HSC氧化應(yīng)激損傷中的作用及機(jī)制[D];第四軍醫(yī)大學(xué);2011年

4 李玉亮;川芎及其有效成分對(duì)照射所致機(jī)體氧化應(yīng)激損傷的保護(hù)作用[D];第四軍醫(yī)大學(xué);2011年

5 王改;姜黃素對(duì)腸單層上皮氧化應(yīng)激損傷的保護(hù)作用[D];河北醫(yī)科大學(xué);2010年

6 王茜;窖蛋白-1在波動(dòng)性高糖誘導(dǎo)的內(nèi)皮細(xì)胞氧化應(yīng)激損傷中的作用及機(jī)制[D];大連醫(yī)科大學(xué);2011年

7 李瓊;Sulfiredoxin-1對(duì)雙氧水誘導(dǎo)的PC12細(xì)胞氧化應(yīng)激損傷的保護(hù)作用研究[D];重慶醫(yī)科大學(xué);2013年

8 張倩;金絲桃苷對(duì)Nrf2-ARE途徑的影響及其對(duì)肝細(xì)胞氧化應(yīng)激損傷的保護(hù)作用[D];西南大學(xué);2014年

9 楊絲絲;硫化氫對(duì)PC12細(xì)胞氧化應(yīng)激損傷的保護(hù)作用及其機(jī)制的探討[D];南華大學(xué);2007年

10 張素響;Nrf2介導(dǎo)促紅細(xì)胞生成素對(duì)神經(jīng)元抗氧化應(yīng)激損傷機(jī)制研究[D];華中科技大學(xué);2013年

本文編號(hào)：2239332