本文選題：原花青素 + 玉米赤霉烯酮　； 參考：《沈陽(yáng)農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：背景:玉米赤霉烯酮(ZEA)又叫作F-2毒素,是一種由鐮孢菌屬真菌產(chǎn)生的具有雌激素活性的非甾醇類(lèi)真菌毒素,與體內(nèi)雌二醇受體具有特殊親和力。ZEA作為一種生物毒素具有特殊的毒性,其毒性作用主要表現(xiàn)在它的強(qiáng)雌激素樣作用,直接對(duì)動(dòng)物機(jī)體生殖系統(tǒng)造成嚴(yán)重?fù)p傷,造成動(dòng)物繁殖功能下降,對(duì)畜牧業(yè)造成嚴(yán)重經(jīng)濟(jì)損失。原花青素(PC)作為一種國(guó)際上公認(rèn)的最有效的清除人體內(nèi)自由基的天然抗氧化劑具有極強(qiáng)的抗氧化作用,能有效清除超氧陰離子自由基和羥基自由基,且其來(lái)源十分廣泛,本試驗(yàn)中選用PC作為ZEA氧化損傷作用的抗氧化劑來(lái)研究PC是否可以有效拮抗ZEA對(duì)鼠睪丸支持細(xì)胞的氧化損傷。目的:本試驗(yàn)旨在研究體外實(shí)驗(yàn)中PC在ZEA誘導(dǎo)的鼠睪丸支持細(xì)胞氧化損傷中的保護(hù)作用及其作用機(jī)制,為畜牧業(yè)生產(chǎn)中ZEA酮中毒的防治提供理論依據(jù)。方法:利用MTT法分別測(cè)出不同濃度的ZEA和PC單獨(dú)和聯(lián)合對(duì)鼠睪丸支持細(xì)胞相對(duì)存活率;流式細(xì)胞術(shù)法檢測(cè)各組支持細(xì)胞凋亡率;利用試劑盒檢測(cè)各組細(xì)胞T-SOD、LDH、GSH-Px活性及MDA和GSH含量;RT-PCR法分別檢測(cè)核因子E2相關(guān)因子2(Nrf2)、血紅素加氧酶1(HO-1)、谷胱甘肽過(guò)氧化物酶(GSH-Px)、醌氧化還原酶1(NQO1)、γ-谷氨酰半胱氨酸合成酶(γ-GCS)的mRNA表達(dá)水平;利用Western-blot法分別檢測(cè)各組細(xì)胞Nrf2、GSH-Px、HO-1、γ-GCS、NQO1的蛋白表達(dá)水平。結(jié)果:1、ZEA對(duì)支持細(xì)胞具有損傷作用,低濃度的PC對(duì)支持細(xì)胞有促進(jìn)生長(zhǎng)的作用,PC對(duì)經(jīng)ZEA染毒的支持細(xì)胞具有保護(hù)作用。2、與ZEA染毒組相比,PC能夠降低經(jīng)ZEA染毒的支持細(xì)胞的凋亡率(p0.01);與ZEA染毒組相比,PC能夠提高T-SOD、GSH-PX活性(p0.05,p0.05),降低MDA含量,顯著降低LDH活性(p0.05),顯著提高GSH含量(p0.05)。3、與ZEA組相比,PC能夠顯著上調(diào)生精細(xì)胞Nrf2及其下游靶基因GSH-Px、HO-1、γ-GCS、NQO1的mRNA和蛋白表達(dá)量。結(jié)論:PC對(duì)ZEA誘導(dǎo)的鼠睪丸支持細(xì)胞氧化損傷具有保護(hù)作用,PC對(duì)ZEA誘導(dǎo)的鼠睪丸支持細(xì)胞損傷的保護(hù)作用可能是通過(guò)激活Nrf2/ARE通路途徑實(shí)現(xiàn)。
[Abstract]:Background: ZEAA, also known as F-2 toxin, is an estrogen-active nonsterol mycotoxin produced by Fusarium fungi.As a biotoxin, ZEA has a special toxicity. Its toxic effect is mainly due to its strong estrogen-like effect, which directly causes serious damage to the reproductive system of animals.Causes the animal reproduction function to decline, causes the serious economic loss to the animal husbandry.As one of the most effective natural antioxidants for scavenging free radicals in human body, proanthocyanidin (PCP) has a strong antioxidant effect, and can effectively scavenge superoxide anion radical and hydroxyl radical, and its sources are very extensive.In this study, PC was used as an antioxidant for oxidative damage of ZEA to investigate whether PC could effectively antagonize oxidative damage of rat testicular Sertoli cells induced by ZEA.Objective: to study the protective effect and mechanism of PC on oxidative injury of testis Sertoli cells induced by ZEA in vitro, and to provide theoretical basis for the prevention and treatment of ZEA ketone poisoning in animal husbandry.Methods: the relative survival rate of rat testicular Sertoli cells with different concentrations of ZEA and PC was determined by MTT method, and the apoptosis rate of Sertoli cells in each group was detected by flow cytometry.The activity of T-SODX LDH-Px and the contents of MDA and GSH were detected by RT-PCR, and the mRNA expression levels of nuclear factor E2 related factor 2Nrf2, heme oxygenase 1HO-1, glutathione peroxidase (GSH-PxX), quinone redox enzyme 1NQO1, 緯 -glutamyl cysteine synthase (緯 -GCSA) were detected by RT-PCR.Western-blot assay was used to detect the protein expression level of Nrf2GCSNQO1, GSH-PxHO-1 and 緯 -GCSO-1 in each group.Results [WT5HZ] ZEA could injure Sertoli cells.Low concentration of PC can promote the growth of Sertoli cells. PC has protective effect on Sertoli cells exposed to ZEA. Compared with ZEA group, it can reduce the apoptosis rate of Sertoli cells exposed to ZEA, and compared with ZEA group, it can reduce the apoptosis rate of Sertoli cells.The activity of GSH-PX in T-SODX was increased, and the activity of GSH-PX was increased. The activity of GSH-PX was increased, and the content of MDA was decreased.Compared with ZEA group, PPC could significantly up-regulate the expression of mRNA and protein in spermatogenic cell Nrf2 and its downstream target genes GSH-PxHO-1, 緯 -GCSNQO1.Conclusion the protective effect of ZEA on oxidative injury of rat testicular Sertoli cells induced by ZEA may be realized by activating the pathway of Nrf2/ARE pathway in ZEA induced rat testicular Sertoli cells injury.
【學(xué)位授予單位】：沈陽(yáng)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)】：S859.87
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本文編號(hào)：1762749