本文選題：茶多酚 + 奶牛��； 參考：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：本論文分四個(gè)部分,主要研究茶多酚對(duì)氧化損傷奶牛乳腺上皮細(xì)胞的干預(yù)作用及機(jī)制。試驗(yàn)一采用奶牛乳腺細(xì)胞構(gòu)建乳腺上皮細(xì)胞系,使其達(dá)到最佳的細(xì)胞系模型。試驗(yàn)二用H_2O_2處理奶牛乳腺上皮細(xì)胞,篩選出H_2O_2最佳誘導(dǎo)劑量和作用時(shí)間,建立氧化應(yīng)激損傷模型。試驗(yàn)三在建立奶牛乳腺上皮細(xì)胞氧化損傷模型的基礎(chǔ)上,添加不同濃度的茶多酚,通過(guò)細(xì)胞培養(yǎng)液中抗氧化指標(biāo)的變化探討茶多酚對(duì)氧化應(yīng)激所致的奶牛乳腺上皮細(xì)胞損傷的影響,并篩選出最佳茶多酚添加劑量。試驗(yàn)四利用特殊性MAPK通路抑制劑分別將p38、JNK、ERK5和ERK1/2四條MAPK通路阻斷,從而篩選出MAPK通路中對(duì)茶多酚影響Nrf2起關(guān)鍵作用的因子,揭示出茶多酚對(duì)奶牛乳腺抗氧化功能的影響機(jī)制。試驗(yàn)結(jié)果如下:(1)本試驗(yàn)成功建立了奶牛乳腺上皮細(xì)胞的體外培養(yǎng)方法,2×10~4個(gè)/mL可以作為以后的細(xì)胞接種濃度。采用免疫熒光染色方法成功鑒定了 BMECs特有骨架蛋白-角蛋白的存在,證明本試驗(yàn)所使用細(xì)胞為BMECs,可用于后續(xù)的試驗(yàn)研究。(2)當(dāng)H_2O_2作用濃度為600μM,作用時(shí)間為6h,乳腺上皮細(xì)胞產(chǎn)生了明顯的氧化損傷,但這種損傷不會(huì)造成細(xì)胞大量死亡,損傷還有可能會(huì)被修復(fù),可作為建立乳腺上皮細(xì)胞氧化損傷模型時(shí)的適宜條件。(3)茶多酚主要通過(guò)抗氧化作用中和細(xì)胞內(nèi)產(chǎn)生的過(guò)多的ROS,提高細(xì)胞內(nèi)Nrf2的表達(dá),減少氧化應(yīng)激,起到保護(hù)乳腺上皮細(xì)胞免受H_2O_2損傷的作用。當(dāng)作用濃度100μg/mL,作用時(shí)間12h時(shí)可作為茶多酚保護(hù)乳腺上皮細(xì)胞產(chǎn)生氧化損傷的最適條件。(4)茶多酚可提高H_2O_2損傷的乳腺上皮細(xì)胞中Nrf2通路和MAPK通路中的基因的表達(dá),阻斷劑DNCB下調(diào)了 MAPK信號(hào)通路中ERK1/2和p38的表達(dá),說(shuō)明TP對(duì)H_2O_2損傷的乳腺上皮細(xì)胞的保護(hù)作用主要是通過(guò)調(diào)節(jié)MAPK通路中ERK1/2和p38的表達(dá)來(lái)促進(jìn)Nrf2的表達(dá)和Nrf2介導(dǎo)的HO-1的表達(dá)。綜上所述,TP對(duì)H202損傷的乳腺上皮細(xì)胞具有保護(hù)作用,其機(jī)制主要是通過(guò)調(diào)節(jié)MAPK通路中ERK1/2和p38的表達(dá)來(lái)促進(jìn)Nrf2和Nrf2介導(dǎo)的HO-1的表達(dá),減緩細(xì)胞受到氧化應(yīng)激造成的損傷,進(jìn)而降低由氧化應(yīng)激造成的疾病的發(fā)生率。
[Abstract]:This paper is divided into four parts to study the effect and mechanism of tea polyphenols on oxidative injury of dairy cow mammary epithelial cells. In experiment 1, the mammary epithelial cell line was constructed by using dairy mammary gland cells to make it the best cell line model. In the second experiment, H_2O_2 was used to treat dairy cow mammary epithelial cells. The optimal dose and time of H_2O_2 induction were screened out, and the oxidative stress injury model was established. On the basis of the model of oxidative injury of dairy cow mammary epithelial cells, different concentrations of tea polyphenols were added in experiment 3. The effects of tea polyphenols on the injury of dairy cow mammary epithelial cells induced by oxidative stress were studied through the changes of antioxidant indexes in cell culture medium. The optimum additive amount of tea polyphenols was screened out. In experiment 4, the specific MAPK pathway inhibitor was used to block the four MAPK pathways of p38 MAPK ERK5 and ERK1/2, respectively, so as to screen out the factors that play a key role in the influence of tea polyphenols on Nrf2 in the MAPK pathway, and to reveal the mechanism of the effects of tea polyphenols on the antioxidant function of dairy cows' mammary glands. The results are as follows: (1) in this experiment, the method of in vitro culture of dairy cow mammary epithelial cells was established successfully. 2 脳 10 ~ 4 / mL of milk cow mammary epithelial cells could be used as the subsequent cell inoculation concentration. The existence of BMECs specific skeleton protein keratin was successfully identified by immunofluorescence staining. It is proved that the cells used in this study are BMECs, which can be used for further experimental study. (2) when the concentration of H_2O_2 is 600 渭 M and the exposure time is 6 h, the epithelial cells of mammary gland produce obvious oxidative damage, but this kind of damage will not result in a large number of cell death. The injury may also be repaired, which can be used as a suitable condition for the establishment of oxidative injury model of breast epithelial cells. Tea polyphenols can increase the expression of Nrf2 and reduce oxidative stress mainly through the antioxidant action and the excessive ROSs produced in the cells. It can protect breast epithelial cells from H_2O_2 damage. When the concentration of tea polyphenols was 100 渭 g / mL and the exposure time was 12 h, tea polyphenols could be used as the best condition for protecting breast epithelial cells from oxidative damage.) Tea polyphenols could increase the expression of Nrf2 pathway and MAPK pathway gene in breast epithelial cells damaged by H_2O_2. The blocking agent DNCB down-regulated the expression of ERK1/2 and p38 in MAPK signaling pathway, suggesting that TP can promote the expression of Nrf2 and Nrf2 mediated HO-1 by regulating the expression of ERK1/2 and p38 in MAPK pathway. In conclusion, TP has protective effect on H202 damaged mammary epithelial cells. The mechanism is to promote the expression of Nrf2 and Nrf2 mediated HO-1 by regulating the expression of ERK1/2 and p38 in the MAPK pathway, and to slow down the damage caused by oxidative stress. This in turn reduces the incidence of diseases caused by oxidative stress.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S823

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 趙文紅;崔慧嫻;孫倩男;陳兵兵;;茶多酚對(duì)直鏈烷基苯磺酸鈉致小鼠皮膚損傷的保護(hù)作用[J];茶葉科學(xué);2016年05期

2 武芳芹;;茶多酚功能及其調(diào)節(jié)脂類(lèi)物質(zhì)代謝機(jī)理研究進(jìn)展[J];現(xiàn)代農(nóng)業(yè)科技;2015年21期

3 陳時(shí)宏;;茶多酚對(duì)動(dòng)物腦組織受自由基損傷的保護(hù)作用研究[J];海峽藥學(xué);2015年09期

4 劉曉慧;揭國(guó)良;林康;吳媛媛;王岳飛;屠幼英;;EGCG和茶氨酸對(duì)細(xì)胞氧化損傷的協(xié)同保護(hù)和修復(fù)作用研究[J];茶葉科學(xué);2014年03期

5 馬蕊;劉仲華;黃建安;林勇;陳金華;鐘源;;綠茶和紅茶提取物抑制中波紫外線誘導(dǎo)HaCaT細(xì)胞氧化損傷和凋亡的比較[J];湖南農(nóng)業(yè)大學(xué)學(xué)報(bào)(自然科學(xué)版);2013年04期

6 丁怡;錢(qián)珂;支楓;王榕;孫欣;;番茄紅素對(duì)過(guò)氧化氫誘導(dǎo)的小鼠來(lái)源神經(jīng)瘤母細(xì)胞損傷的保護(hù)作用及機(jī)制研究[J];中華老年心腦血管病雜志;2013年06期

7 陳浩;許浪;;Nrf2/NQO1信號(hào)通路在胃癌干細(xì)胞氧化應(yīng)激反應(yīng)中的作用機(jī)制[J];吉林大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2013年03期

8 姜敏;高振;李風(fēng)森;;Keap1-Nrf2-ARE通路與慢性阻塞性肺疾病氧化/抗氧化失衡關(guān)系[J];國(guó)際病理科學(xué)與臨床雜志;2013年02期

9 王釗;黃雨蒙;彭軍;;氧化應(yīng)激與腦缺血-再灌注損傷[J];國(guó)際病理科學(xué)與臨床雜志;2012年04期

10 王振云;周璇;李惠俠;廖旭東;崔群維;李蓮;王根林;;茶多酚對(duì)氧化應(yīng)激所致奶牛乳腺上皮細(xì)胞損傷的保護(hù)作用[J];南京農(nóng)業(yè)大學(xué)學(xué)報(bào);2012年03期

相關(guān)博士學(xué)位論文 前5條

1 楊光照;EGCG通過(guò)上調(diào)Nrf2/HO-1途徑降低PM2.5所致HUVEC氧化損傷的機(jī)制研究[D];山西醫(yī)科大學(xué);2015年

2 金鹿;維生素A對(duì)奶牛乳腺硒蛋白合成及抗氧化功能影響機(jī)理的研究[D];內(nèi)蒙古農(nóng)業(yè)大學(xué);2014年

3 堅(jiān)哲;Nrf2-ARE信號(hào)通路在白癜風(fēng)氧化應(yīng)激發(fā)病中的作用和機(jī)制研究[D];第四軍醫(yī)大學(xué);2013年

4 李永義;茶多酚對(duì)氧化應(yīng)激仔豬的保護(hù)作用及機(jī)制研究[D];四川農(nóng)業(yè)大學(xué);2011年

5 王艷明;日糧脂肪和能量水平對(duì)奶牛氧化應(yīng)激、生產(chǎn)性能的影響及抗氧化劑添加效果研究[D];浙江大學(xué);2010年

相關(guān)碩士學(xué)位論文 前3條

1 姜婧;原花青素對(duì)鉛誘導(dǎo)的大鼠肝毒性的保護(hù)作用研究[D];東北農(nóng)業(yè)大學(xué);2016年

2 林佳;番茄紅素通過(guò)Nrf2信號(hào)通路拮抗阿特拉津致小鼠腎毒性的機(jī)制[D];東北農(nóng)業(yè)大學(xué);2016年

3 劉志方;葉黃素基于Nrf-2/ARE信號(hào)途徑對(duì)人結(jié)腸癌HT29細(xì)胞增殖抑制作用的研究[D];鄭州大學(xué);2016年

，

本文編號(hào)：1941483