本文選題：鉛 + EGCG��； 參考：《鄭州大學(xué)》2017年碩士論文

【摘要】：目的研究表沒(méi)食子兒茶素沒(méi)食子酸酯(epigallocatechin gallate,EGCG)與維生素E(vitamin E,VitE)聯(lián)用對(duì)不同時(shí)期母鼠鉛暴露所致仔鼠阿爾茨海默病(Alzheimer’s disease,AD)淀粉樣變的干預(yù)作用,探討EGCG與VitE的聯(lián)用對(duì)鉛致AD樣病變是否存在協(xié)同作用及其可能機(jī)制。方法將SPF級(jí)昆明小鼠33只(雌22只、雄11只),按照體重順序隨機(jī)分為3組:對(duì)照組(雌2只、雄1只)、孕前期染鉛組(雌10只、雄5只)和哺乳期染鉛組(雌10只、雄5只)。孕前期染鉛組雌鼠飲0.2%醋酸鉛溶液染鉛21d,染鉛結(jié)束后與該組雄鼠合籠以獲得孕鼠,自由飲去離子水直至仔鼠斷乳。哺乳期染鉛組正常飲去離子水、合籠,待孕鼠分娩后飲0.2%醋酸鉛溶液染鉛21d直至小鼠斷乳。對(duì)照組在實(shí)驗(yàn)過(guò)程中自由飲去離子水,合籠后獲得孕鼠直至分娩后仔鼠斷乳。斷乳后,孕前期染鉛組和哺乳期染鉛組所產(chǎn)仔鼠再按體重順序編號(hào),分別隨機(jī)分為7組,即生理鹽水組、玉米油組、EGCG干預(yù)組(3.0 mg/kg)、VitE干預(yù)組(30 IU/kg)、聯(lián)合干預(yù)組(1)(EGCG 3.0 mg/kg+VitE 30 IU/kg)、聯(lián)合干預(yù)組(2)(EGCG 1.5 mg/kg+VitE 60 IU/kg)、聯(lián)合干預(yù)組(3)(EGCG 6.0 mg/kg+Vit E 15 IU/kg),每組14只,從21 d斷乳后開(kāi)始連續(xù)灌胃28 d。對(duì)照組所產(chǎn)仔鼠按體重順序隨機(jī)選取14只自由飲去離子水。干預(yù)結(jié)束后,對(duì)所有小鼠進(jìn)行Morris水迷宮實(shí)驗(yàn);用極譜儀測(cè)定小鼠血鉛含量;用相應(yīng)試劑盒測(cè)定小鼠海馬組織中總超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(malondialdehvde,MDA)含量;用ELISA試劑盒檢測(cè)小鼠海馬組織中Aβ_(1-40)、Aβ_(1-42)含量;用Western blot檢測(cè)小鼠海馬組織中β淀粉樣前體蛋白(β-amyloid precursor protein,APP)及β-內(nèi)分泌酶1(β-secreatase-1,BACE1)蛋白的表達(dá)水平。使用SPSS 21.0版本進(jìn)行統(tǒng)計(jì)分析,檢驗(yàn)水準(zhǔn)α=0.05。結(jié)果1.體重結(jié)果顯示:孕前期染鉛組和哺乳期染鉛組的小鼠,與對(duì)照組相比,第5周齡開(kāi)始生理鹽水組、玉米油組及不同干預(yù)組小鼠體重均明顯降低(P0.05);第9周齡時(shí)孕前期染鉛組中聯(lián)合干預(yù)組(1)和聯(lián)合干預(yù)組(3)的小鼠體重分別高于生理鹽水組和玉米油組(P0.05),哺乳期染鉛組中聯(lián)合干預(yù)組(1)的小鼠體重高于玉米油組(P0.05)。2.水迷宮結(jié)果顯示:孕前期染鉛組和哺乳期染鉛組中,生理鹽水組和玉米油組的平均逃避潛伏期均高于對(duì)照組(P0.05);EGCG干預(yù)組和VitE干預(yù)組的平均逃避潛伏期分別低于生理鹽水組和玉米油組(P0.05);與單獨(dú)干預(yù)組相比,孕前期染鉛組中聯(lián)合干預(yù)組(1)的平均逃避潛伏期降低(P0.05);哺乳期染鉛組中聯(lián)合干預(yù)組(1)和(3)的平均逃避潛伏期降低(P0.05)。孕前期和哺乳期染鉛組中,生理鹽水組和玉米油組的穿越平臺(tái)次數(shù)均明顯低于對(duì)照組(P0.05);三個(gè)聯(lián)合干預(yù)組分別與生理鹽水組和玉米油組相比,穿越平臺(tái)次數(shù)明顯增加(P0.05);與單獨(dú)干預(yù)組相比,聯(lián)合干預(yù)組(1)和(3)的穿越平臺(tái)次數(shù)明顯增加(P0.05)。3.血鉛含量結(jié)果顯示:孕前期染鉛組和哺乳期染鉛組中,生理鹽水組和玉米油組的血鉛含量均明顯高于對(duì)照組(P0.05);EGCG干預(yù)組和三個(gè)聯(lián)合干預(yù)組血鉛含量均低于生理鹽水組(P0.05);三個(gè)聯(lián)合干預(yù)組血鉛含量均低于玉米油組(P0.05),而單獨(dú)干預(yù)組與聯(lián)合干預(yù)組的血鉛含量差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。4.氧化應(yīng)激指標(biāo)結(jié)果顯示:孕前期染鉛組和哺乳期染鉛組中,生理鹽水組和玉米油組與對(duì)照組相比,SOD活性顯著降低(P0.05),MDA含量明顯升高(P0.05);分別與生理鹽水組和玉米油組相比,單獨(dú)干預(yù)組和三個(gè)聯(lián)合干預(yù)組的SOD活性升高(P0.05),MDA含量降低(P0.05);聯(lián)合干預(yù)組與單獨(dú)干預(yù)組相比,聯(lián)合干預(yù)組(1)、(3)的SOD活性升高(P0.05),MDA含量差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。5.Aβ_(1-40)、Aβ_(1-42)含量結(jié)果顯示:孕前期染鉛組和哺乳期染鉛組中,生理鹽水組和玉米油組Aβ_(1-40)、Aβ_(1-42)含量均明顯高于對(duì)照組(P0.05);與生理鹽水組和玉米油組相比,單獨(dú)干預(yù)組和聯(lián)合干預(yù)組的Aβ_(1-40)、Aβ_(1-42)含量降低(P0.05);與單獨(dú)干預(yù)組相比,哺乳期染鉛組中,聯(lián)合干預(yù)組(3)Aβ_(1-42)含量比VitE干預(yù)組低(P0.05)。6.APP、BACE1蛋白表達(dá)水平結(jié)果顯示:孕前期染鉛組和哺乳期染鉛組中,與對(duì)照組相比,生理鹽水組、玉米油組及不同干預(yù)組APP、BACE1蛋白水平明顯增多(P0.05);且不同干預(yù)組與生理鹽水組和玉米油組相比,APP、BACE1蛋白水平明顯減少(P0.05);聯(lián)合干預(yù)組與單獨(dú)干預(yù)組相比,APP、BACE1蛋白水平明顯減少(P0.05)。結(jié)論1.母鼠孕前期或哺乳期的鉛暴露,可導(dǎo)致小鼠體內(nèi)血鉛含量升高,影響小鼠生長(zhǎng)發(fā)育,損傷小鼠空間學(xué)習(xí)記憶能力,并且可以升高海馬組織中氧化應(yīng)激水平,誘導(dǎo)海馬中APP和BACE1蛋白的過(guò)度表達(dá),從而增加海馬中Aβ_(1-40)、Aβ_(1-42)的水平。2.EGCG、VitE以及EGCG和VitE聯(lián)用均可以改善鉛暴露引起的小鼠學(xué)習(xí)記憶能力下降情況,降低AD樣變小鼠海馬中Aβ_(1-40)、Aβ_(1-42)的含量,降低APP及BACE1蛋白的表達(dá)水平,并且在適當(dāng)劑量下的聯(lián)合干預(yù)效果要優(yōu)于單獨(dú)使用EGCG或VitE。
[Abstract]:Objective to investigate the effect of combined use of epigallocatechin gallate (EGCG) and vitamin E (vitamin E, VitE) on amyloidosis of Alzheimer's disease (Alzheimer 's disease, AD) induced by lead exposure in mice at different times. Methods 33 Kunming mice (22 females and 11 males) were randomly divided into 3 groups according to the weight sequence: the control group (2 female, 1 male), the prepregnancy lead group (10 female, male 5) and the lactating group (10, male 5). The female rats in the pre pregnancy group were exposed to lead 21d in the lead group, and the lead was closed with the male mice. In the cage, the pregnant rats were free to drink deionized water until the offspring were weaned. The lactating group normally drank deionized water and closed the cage. After childbirth, 0.2% lead acetate solution was used to dye lead 21d until the mice were weaned. The control group was free to drink deionized water during the experiment. The offspring rats in the group and lactation stage were numbered according to the weight sequence and were randomly divided into 7 groups, namely the saline group, the corn oil group, the EGCG intervention group (3 mg/kg), the VitE intervention group (30 IU/kg), the combined intervention group (1) (EGCG 3 mg/kg+VitE 30 IU/kg), the combined intervention group (2) (EGCG 1.5 mg/kg+VitE 60 IU/kg), and the joint intervention group (3) (EGCG 6 mg/kg+Vi). T E 15 IU/kg), 14 rats in each group, 14 free drinking deionized water were selected randomly from 28 d. control group after 21 d weaning. After the intervention, all mice were tested in the Morris water maze test; the blood lead content of mice was measured by polarography; and the total superoxide dismutase in the hippocampus of mice was measured by the phase strain kit. Superoxide dismutase (SOD) activity and the content of malondialdehyde (malondialdehvde, MDA), and the content of A beta (1-40) and A beta (1-42) in the hippocampus of mice were detected by ELISA kit, and the beta amyloid precursor protein (beta -amyloid precursor) and beta endocrine enzyme 1 protein in the hippocampus of mice were detected by Western blot. The level of expression of SPSS 21 was statistically analyzed, and the results of test level alpha =0.05. results showed that the results of 1. body weight showed that the mice in the prepregnancy group and the breastfeeding group, compared with the control group, began to start the physiological saline group at fifth weeks of age, and the weight of the corn oil group and the different intervention groups were significantly decreased (P0.05), and the lead exposure group at the early stage of pregnancy at the age of ninth weeks was the lead group. The weight of mice in the combined intervention group (1) and the combined intervention group (3) was higher than that of the saline group and the corn oil group (P0.05). The weight of the mice in the combined intervention group (1) in the lactating period (1) was higher than that of the corn oil group (P0.05).2. water maze. The incubation period was higher than that of the control group (P0.05); the average escape latency of the EGCG intervention group and the VitE intervention group was lower than that of the saline group and the corn oil group (P0.05). Compared with the single intervention group, the average escape latency decreased (P0.05) in the combined intervention group (1) in the prepregnancy group, and the average escape of the combined intervention group (1) and (3) in the breastfeeding lead dyed lead group. The number of crossing platforms in the saline group and the corn oil group were significantly lower than that of the control group (P0.05) in the preconception and lactation period (P0.05). The number of crossing platforms increased significantly (P0.05) compared with the saline group and the corn oil group (1) and (3) in the combined intervention group (P0.05). The blood lead content of the crossing platform (P0.05).3. showed that the blood lead content of the normal saline group and the corn oil group was significantly higher than that of the control group (P0.05), the blood lead content in the EGCG intervention group and the three combined intervention group was lower than that of the normal saline group (P0.05), and the blood lead content in the three combined intervention group was more than that of the control group. The blood lead content of the group was lower than that of the corn oil group (P0.05), but there was no significant difference in the blood lead content between the single intervention group and the combined intervention group (P0.05). The results of.4. oxidative stress showed that the activity of SOD was significantly decreased (P0.05) and the content of MDA significantly increased (P0.05) compared with the control group. Compared with the normal saline group and the corn oil group, the activity of SOD increased (P0.05) and the content of MDA decreased (P0.05) in the individual intervention group and the three combined intervention group. The combined intervention group was compared with the single intervention group, and the SOD activity in the combined intervention group (1) and (3) increased (P0.05), and the MDA content was not statistically significant (P0.05).5.A beta (1-40), and A beta (1-42) content results. The contents of A beta (1-40) and A beta (1-42) in the normal saline group and the corn oil group were significantly higher than those in the control group (P0.05). Compared with the normal saline group and the corn oil group, the content of A beta (1-40) and the A beta (1-42) in the single intervention group and the combined intervention group were lower (P0.05). Compared with the single intervention group, the lactation period lead dye group was compared with the single intervention group. In the combined intervention group (3), the content of A beta (1-42) was lower than that of the VitE intervention group (P0.05).6.APP, and the expression level of BACE1 protein showed that the level of BACE1 egg white in the normal saline group, the corn oil group and the intervention group APP, and the BACE1 egg white level increased significantly (P0.05), and the different intervention groups and the saline group and the jade were compared with the control group. Compared with the rice oil group, the level of APP and BACE1 protein decreased significantly (P0.05), and the level of APP and BACE1 protein decreased significantly in the combined intervention group compared with the single intervention group (P0.05). Conclusion the lead exposure in the prepregnancy or lactation period of the 1. female mice could lead to the increase of blood lead content in the mice, the growth and development of mice and the ability of learning and memory in mice. To increase the level of oxidative stress in hippocampus, induced overexpression of APP and BACE1 protein in hippocampus, thus increasing A beta (1-40) and A beta (1-42) level.2.EGCG, VitE and EGCG and VitE can improve the decline of learning and memory ability in mice induced by lead exposure, and reduce A beta (1-40) and A beta (1-42) in the hippocampus of the AD like mice. The level of APP and BACE1 protein was decreased, and the combined intervention at appropriate dose was better than using EGCG or VitE. alone.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R749.16

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 李靜;劉芳麗;安偉鋒;薛振菲;李文杰;;不同時(shí)期鉛暴露對(duì)小鼠學(xué)習(xí)記憶能力及海馬組織中β淀粉樣蛋白表達(dá)的影響[J];鄭州大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2015年03期

2 薛振菲;劉芳麗;李靜;李寧;尹延彥;李文杰;;鉛對(duì)小鼠學(xué)習(xí)記憶能力及β-淀粉樣前體蛋白表達(dá)的影響[J];現(xiàn)代預(yù)防醫(yī)學(xué);2014年15期

3 邱宏;金國(guó)琴;金如鋒;趙偉康;;水迷宮重復(fù)測(cè)量數(shù)據(jù)的方差分析及其在SPSS中的實(shí)現(xiàn)[J];中西醫(yī)結(jié)合學(xué)報(bào);2007年01期

4 劉英莉,張艷淑,王廣增,姚林,趙伯陽(yáng);茶多酚聯(lián)合維生素C、維生素E對(duì)染塵大鼠肺泡巨噬細(xì)胞DNA損傷的保護(hù)實(shí)驗(yàn)研究[J];中國(guó)職業(yè)醫(yī)學(xué);2000年05期

5 張艷淑,王廣增,姚林,趙伯陽(yáng),劉英莉;茶多酚聯(lián)合維生素C、E對(duì)染塵大鼠血液中NO、NOS和GSH-Px的影響[J];中國(guó)公共衛(wèi)生;2000年10期

，

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