本文選題：茶葉　切入點(diǎn)：EGCG　出處：《湖南農(nóng)業(yè)大學(xué)》2015年博士論文　論文類(lèi)型：學(xué)位論文

【摘要】：表沒(méi)食子兒茶素沒(méi)食子酸酯(Epigallocatechin-3-gallate, EGCG)是綠茶中主要的兒茶素類(lèi)物質(zhì),具有抗氧化、抗過(guò)敏、抗炎癥、抗輻射和降血脂等一系列生物活性。EGCG對(duì)活體壽命的影響研究較少且存在爭(zhēng)議。本文在前人已有研究基礎(chǔ)上,進(jìn)一步探討EGCG對(duì)秀麗線(xiàn)蟲(chóng)(Caenorhabditis elegans)壽命的調(diào)節(jié)作用及其潛在作用機(jī)制。主要研究結(jié)果如下：(1)通過(guò)EGCG干預(yù)野生型秀麗線(xiàn)蟲(chóng)N2整個(gè)成蟲(chóng)期的壽命試驗(yàn)發(fā)現(xiàn),在0-300 μM濃度范圍內(nèi)與秀麗線(xiàn)蟲(chóng)平均壽命正相關(guān)；在300-500 μM范圍內(nèi)不顯著影響秀麗線(xiàn)蟲(chóng)平均壽命；在濃度超過(guò)500μM時(shí)與秀麗線(xiàn)蟲(chóng)的平均壽命負(fù)相關(guān)。不同食物結(jié)構(gòu)(活菌、死菌和EGCG預(yù)處理大腸桿菌)壽命試驗(yàn)發(fā)現(xiàn),EGCG調(diào)節(jié)秀麗線(xiàn)蟲(chóng)壽命與其抗菌性無(wú)關(guān)；同時(shí)添加EGCG和過(guò)氧化物酶(CAT)進(jìn)行壽命試驗(yàn)發(fā)現(xiàn),EGCG調(diào)節(jié)秀麗線(xiàn)蟲(chóng)壽命與其在NGM中自動(dòng)氧化生成的H202無(wú)關(guān)。(2)通過(guò)EGCG限時(shí)干預(yù)野生型秀麗線(xiàn)蟲(chóng)N2成蟲(chóng)早期和中期(6 d)、干預(yù)不同年齡階段秀麗線(xiàn)蟲(chóng)(成蟲(chóng)第6、12和18d開(kāi)始干預(yù))和干預(yù)秀麗線(xiàn)蟲(chóng)整個(gè)成蟲(chóng)期對(duì)其衰老相關(guān)功能衰退的影響發(fā)現(xiàn),EGCG調(diào)節(jié)秀麗線(xiàn)蟲(chóng)壽命的主要作用時(shí)期是成蟲(chóng)早中期。通過(guò)去除NGM中蛋白胨進(jìn)行野生型秀麗線(xiàn)蟲(chóng)N2壽命試驗(yàn)發(fā)現(xiàn),EGCG對(duì)秀麗線(xiàn)蟲(chóng)N2壽命的影響程度較正常培養(yǎng)條件下小。通過(guò)DA465 (eat-2缺失)突變體壽命試驗(yàn)發(fā)現(xiàn),200和1000 μM對(duì)DA465突變體壽命的影響較其對(duì)N2的影響要小。(3)EGCG干預(yù)野生型秀麗線(xiàn)蟲(chóng)N248 h后進(jìn)行應(yīng)激試驗(yàn)發(fā)現(xiàn),在不同應(yīng)激環(huán)境下,不同EGCG濃度產(chǎn)生不同效果,最優(yōu)濃度并不一致。EGCG調(diào)節(jié)秀麗線(xiàn)蟲(chóng)熱應(yīng)激抵抗的劑量效應(yīng)與調(diào)節(jié)壽命一致。200 μM EGCG限時(shí)干預(yù)(6 d)可幫助秀麗線(xiàn)蟲(chóng)N2抵御1000μM EGCG引起的壽命損傷。(4) EGCG不影響秀麗線(xiàn)蟲(chóng)N2生殖和幼蟲(chóng)發(fā)育。400和1000 μM EGCG干預(yù)秀麗線(xiàn)蟲(chóng)幼蟲(chóng)期顯著縮短其體長(zhǎng)。EGCG干預(yù)秀麗線(xiàn)蟲(chóng)N2幼蟲(chóng)期并不影響Fl、F2和F6代(連續(xù)干預(yù)5代)成蟲(chóng)壽命。(5) EGCG(200和1000μM)干預(yù)降低秀麗線(xiàn)蟲(chóng)ATP水平和TG含量,并短期提升ROS含量,激活抗氧化防御系統(tǒng)清除ROS。同時(shí)添加EGCG和抗氧化劑N-乙酰半胱氨酸(NAC)或叔丁基-4-羥基茴香醚(BHA)進(jìn)行壽命試驗(yàn)發(fā)現(xiàn),EGCG(200和1000μM)不再顯著影響野生型秀麗線(xiàn)蟲(chóng)N2壽命。200 μM EGCG提升秀麗線(xiàn)蟲(chóng)NAD+水平,1000 μM EGCG降低秀麗線(xiàn)蟲(chóng)NAD+水平。200 μMEGCG延長(zhǎng)秀麗線(xiàn)蟲(chóng)壽命與AAK-2、 SIR2.1、PRDX-2、PMK-1、DAF-16、SKN-1以及線(xiàn)粒體電子傳遞鏈相關(guān),與HIF-1、 SOD-3和胰島素及類(lèi)胰島素信號(hào)通路無(wú)關(guān)。另外,1000 μM EGCG縮短秀麗線(xiàn)蟲(chóng)亦與SIR2.1相關(guān)。通過(guò)測(cè)定mtDNA與nDNA比例發(fā)現(xiàn),200 μM EGCG能有效促進(jìn)線(xiàn)粒體生物合成。通過(guò)綠色熒光蛋白融合表達(dá)hsp-6和hsp-60轉(zhuǎn)基因品系的綠色熒光表達(dá)及qPCR試驗(yàn)發(fā)現(xiàn),EGCG并未誘導(dǎo)線(xiàn)粒體未折疊蛋白質(zhì)響應(yīng)。
[Abstract]:Epigallocatechin-3-gallateEpigallocatechin-3-gallateEpigallocatechin-3-gallateEpigallocatechin-3-gallateEpigallocatechin-3-gallateEpigallocatechin-3-gallateEpigallocatechin-3-gallateEpigallocatechin-3-gallate (. A series of biological activities, such as anti-radiation and hypolipidemia, have little or no research on the effects of EGCG on life span in vivo. The effects of EGCG on the life span of Caenorhabditis eleans were further studied. The main results were as follows: 1) the results of EGCG intervention on the life span of wild-type nematode N _ 2 in the whole adult stage were as follows:. In the range of 0-300 渭 M, there was a positive correlation with the average life span of the nematode, but not in the range of 300-500 渭 M, but negative correlation with the average life span of the nematode when the concentration was above 500 渭 M. different food structure (living bacteria, The life test of dead bacteria and Escherichia coli pretreated with EGCG showed that there was no relationship between the life span of EGCG-regulated nematode and its antimicrobial activity. EGCG and peroxidase (EGCG) were added in the longevity test. It was found that EGCG-regulated nematode longevity was independent of H202, which was produced automatically in NGM.) EGCG limited time was used to intervene in the early and middle stage of N _ 2 adult nematode wild-type nematode N _ 2 for 6 days. The effects of different age stages on senescence related function of nematodes (beginning intervention on the 12th and 18th days of adult worms) and the whole adult stage of C. elegans showed that EGCG mainly regulates the longevity of C. elegans in the early stage of adults. In the middle stage, by removing peptone from NGM, the N _ 2 longevity of wild type nematode nematode was tested. It was found that the effect of EGCG on N _ 2 longevity of nematode was less than that under normal culture condition. The effect of 1 000 渭 M on the lifespan of DA465 mutant was less than that on N2. The stress test showed that 1 000 渭 M treated wild-type nematode N248 h later. Under different stress conditions, different concentrations of EGCG produced different effects. The dose effect of EGCG regulating the heat stress resistance of nematode nematode is the same as that of the control life. 200 渭 M EGCG for 6 days) can help N 2 to resist 1 000 渭 M EGCG induced life damage. 4) EGCG does not affect N 2 of nematode. Reproductive and larval development. 400 and 1 000 渭 M EGCG intervention significantly shortened the larval stage of nematode nematode. EGCG did not affect the lifespan of F 2 and F 6 generations (5 successive generations) EGCG(200 and 1 000 渭 M). ATP level and TG content, And increase the ROS content in a short time, Activation of antioxidant defense system to remove ros.An addition of EGCG and the antioxidant N-acetylcysteine or tert-#china_person0# -4- hydroxyanisole (BHA) were used to test the life of wild nematode nematode. It was found that EGCG 200 and 1000 渭 M) no longer significantly affected the N2 life span of wild nematode nematode. 200 渭 M. The increase of NAD level by EGCG and the decrease of NAD level by 200 渭 MEGCG were associated with AAK-2, SIR2.1% PRDX-2 PMK-1, DAF-16SKN-1 and mitochondrial electron transport chain. It was not related to HIF-1, SOD-3, insulin and insulin-like signal transduction pathway. The other 1 000 渭 M EGCG shortening nematode was also associated with SIR2.1. It was found that the ratio of mtDNA to nDNA could effectively promote mitochondrial biosynthesis. The green fluorescence expression of hsp-6 and hsp-60 transgenic lines expressed by white fusion and qPCR test showed that EGCG did not induce the response of mitochondrial unfolded protein.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R151

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