發(fā)布時間：2018-05-14 08:22

  本文選題：氯化鎘 + 槲皮素��； 參考：《沈陽農(nóng)業(yè)大學》2017年碩士論文

【摘要】：目的:本試驗探究抗氧化劑槲皮素通過調(diào)控Nrf2-Keap1途徑拮抗鎘對大鼠生殖機能造成的應激損傷。通過體內(nèi)試驗對其機制進行深入研究,為研究鎘的生殖毒性及槲皮素通過調(diào)控Nrf2-Keap1路徑保護機體免受氧化損傷的研究奠定了基礎。方法:通過分組飼養(yǎng),定時灌胃處理,建立試驗模型,4周之后取出睪丸組織,通過檢驗睪丸組織的谷胱甘肽過氧化酶(Gsh-Px),過氧化氫酶(CAT),超岐物過氧化物酶(SOD)等抗氧化酶水平,睪酮分泌量以及Nrf2-Keap1抗氧化通路相關因子2(Nrf2)、谷胱甘肽過氧化物酶(Gsh-Px)、血紅素加氧酶1(HO-1)、Y-谷氨酰半胱氨酸合成酶(γ-GCS)、醌氧化還原酶1(NQO1)mRNA及蛋白的表達量,進一步判定槲皮素作為抗氧化劑對染鎘的干預作用。結果:1.通過對大鼠灌胃處理后,5天作為一個周期,對大鼠體重進行測定,發(fā)現(xiàn)染鎘組大鼠體重明顯低于空白組及槲皮素組,槲皮素組的體重和空白組的體重相比差異顯著。槲皮素治療組的大鼠體重測定值明顯高于染鎘組。2.通過對睪丸組織內(nèi)必要的抗氧化酶檢測,對試驗數(shù)據(jù)進行分析發(fā)現(xiàn),與空白組相比,染鎘組丙二醛(MDA)活性明顯升高(p0.01),差異極顯著,槲皮素組丙二醛活性低于空白組(p0.01),槲皮素治療組與染鎘組比較,丙二醛活性低于染鎘組(p0.01)。多岐物過氧化物酶(SOD)活性,過氧化氫酶(CAT),谷胱甘肽過氧化物酶(Gsh-Px),染鎘組與實驗空白組相比,活性明顯降低(p0.01),槲皮素治療組與染鎘組相比,抗氧化酶的活性明顯升高,差異極顯著(p0.01)。數(shù)據(jù)表明槲皮素對鎘引起的機體損傷具有很好的緩和作用,促使機體的抗氧化反應能力增強。3.通過Elisa酶聯(lián)免疫吸附試驗測定發(fā)現(xiàn),與空白組作比較,染鎘組睪酮含量明顯降低(p0.01),槲皮素組的睪酮含量與空白組相比明顯增加(p0.01),槲皮素治療組與染鎘組相比,睪酮含量明顯上升,差異顯著(p0.05)。證明槲皮素能夠拮抗鎘引起機體睪酮分泌量的減少,減弱對機體的損傷。4.通過對睪丸組織的HE染色鏡檢分析,發(fā)現(xiàn)與空白組相比,染鎘組睪丸組織存在生精上皮生殖細胞大量脫落,各級生殖細胞數(shù)量減少,曲細精管中央伸長型精子細胞數(shù)量減少的現(xiàn)象,槲皮素治療組部分生精上皮細胞損傷脫落,內(nèi)皮細胞損傷情況不顯著,這表示槲皮素對鎘引起的大鼠睪丸組織損傷能夠起到較好的緩解作用。5.通過q-PCR技術和Western Blot蛋白技術檢測組織Nrf2抗氧化通路中Nrf2,HO-1,NQO1,Gsh-Px,γ-GCS,Keap1的mRNA和蛋白水平,發(fā)現(xiàn)與空白組相比,染鎘組Keap1的mRNA和蛋白水平高于空白組(p0.01),然而Nrf2,HO-1,NQO1,Gsh-Px,y-GCS的mRNA和蛋白水平明顯低于空白組(p0.01,p0.05)。對于槲皮素治療組,Keap1的表達水平明顯低于染鎘組(p0.01),Nrf2,HO-1,NQO1,Gsh-Px,y-GCS水平高于染鎘組(p0.01,p0.05)。結論:鎘會損害大鼠睪丸組織的生精功能,降低睪丸組織對抗氧化反應的能力,造成生殖系統(tǒng)明顯損傷,而槲皮素具有拮抗鎘誘導損害作用的能力。槲皮素可以激活Nrf2-Keap1抗氧化通路,調(diào)節(jié)抗氧化酶的表達水平,干預鎘損傷,證明槲皮素對鎘造成的內(nèi)源性氧化反應起到防御功能。
[Abstract]:Objective: To explore the antagonistic effect of quercetin on the stress damage caused by cadmium on the reproductive function of rats by regulating the Nrf2-Keap1 pathway. Through in vivo experiments, the mechanism of quercetin was studied in order to study the reproductive toxicity of cadmium and the study of quercetin to protect the body from oxidative damage by regulating the Nrf2-Keap1 path. Methods: the experimental model was established by grouping and feeding the stomach at a time. After 4 weeks, the testicular tissue was removed, and the levels of antioxidant enzymes, such as glutathione peroxidase (Gsh-Px), catalase (CAT), superoxide peroxidase (SOD), and Nrf2-Keap1 antioxidant pathway related factor 2 (Nrf2), were examined, Gu Guanggan Peptide peroxidase (Gsh-Px), heme oxygenase 1 (HO-1), Y- glutamyl cysteine synthetase (gamma -GCS), quinone oxidoreductase 1 (NQO1) mRNA and protein expression, further determined that quercetin was used as an antioxidant to interfere with cadmium. The results were as follows: 1. of rats were treated by gavage for 5 days as a cycle, and the weight of rats was measured. It was found that the weight of rats in the cadmium dyed group was significantly lower than that in the blank group and the quercetin group. The weight of the quercetin group was significantly different from that in the blank group. The weight determination of the rats in the quercetin group was significantly higher than that in the cadmium dyed group.2. through the test of the necessary antioxidant enzymes in the testicular tissue, and the test data were analyzed and compared with the blank group. The activity of malondialdehyde (MDA) in cadmium dyed group was significantly higher (P0.01). The activity of malondialdehyde in quercetin group was lower than that in blank group (P0.01). The activity of malondialdehyde in quercetin group and cadmium dyed group was lower than that of cadmium dyed group (P0.01). The activity of peroxidase (SOD), catalase (CAT), glutathione peroxidase (Gsh-Px), cadmium dyed group and solid of cadmium group were compared with that of cadmium dyed group. Compared with the blank group, the activity of the quercetin was significantly lower (P0.01). The activity of antioxidant enzymes in the quercetin group was significantly higher than that in the cadmium dyed group, and the difference was very significant (P0.01). The data showed that quercetin had a good mitigating effect on the damage caused by cadmium and enhanced the anti oxygenation ability of the organism by the Elisa enzyme-linked immunosorbent assay. Compared with the blank group, the testosterone content in the cadmium dyed group decreased significantly (P0.01), and the testosterone content in the quercetin group increased significantly compared with the blank group (P0.01). The content of testosterone in the quercetin group was significantly higher than that in the cadmium dyed group (P0.05). It was proved that quercetin could antagonize the decrease of the excretion of testosterone in the organism and the mechanism of reducing the mechanism of cadmium. The damage of.4. was analyzed by the HE staining microscopy of the testicular tissue. It was found that compared with the blank group, the testicular tissue in the cadmium dyed group had a large number of germ cells falling off, the number of germ cells at all levels decreased, the number of central elongated spermatozoa in the fine tubule decreased, and the injury of some spermatogenic epithelial cells in the quercetin group was lost. The damage of skin cells was not significant, which indicated that quercetin could play a better role in alleviating the damage of testicular tissue in rats induced by cadmium..5., HO-1, NQO1, Gsh-Px, gamma -GCS, Keap1 mRNA and protein levels were detected by q-PCR technology and Western Blot protein technology. The level of mRNA and protein was higher than that in the blank group (P0.01), but the level of mRNA and protein in Nrf2, HO-1, NQO1, Gsh-Px, y-GCS was significantly lower than that in the blank group (P0.01, P0.05). Quercetin can activate Nrf2-Keap1 antioxidant pathway, regulate the expression level of antioxidant enzymes, interfere with the cadmium damage, and demonstrate the endogenous oxidation caused by quercetin to cadmium. The reaction serves as a defensive function.

【學位授予單位】：沈陽農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R114

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