發(fā)布時(shí)間：2018-03-22 21:31

  本文選題：卵巢　切入點(diǎn)：半乳糖致衰　出處：《內(nèi)蒙古大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：卵巢是女性生殖系統(tǒng)中最重要的器官之一,隨著婦女年齡的增加,其功能不斷下降,并經(jīng)歷卵泡減少、卵母細(xì)胞質(zhì)量下降、月經(jīng)不調(diào)、卵巢激素缺乏、停止排卵最后導(dǎo)致生育能力下降的過程。而延緩卵巢老化是增加生殖能力的重要手段。藻藍(lán)蛋白(C-Phycocyanin,PC)是來自鈍頂螺旋藻的一種具有抗炎癥、抗氧化、抗癌癥、提高免疫能力和保肝護(hù)腎作用的蛋白質(zhì),因?yàn)樗哂卸喾N生物活性而被應(yīng)用在許多臨床疾病研究中。在前期的研究中,我們已經(jīng)發(fā)現(xiàn)PC具有改善D-半乳糖致衰小鼠卵巢功能的作用,但PC發(fā)揮這一作用的分子機(jī)理還未被充分研究。本研究主要對(duì)D-半乳糖致衰小鼠灌胃藻藍(lán)蛋白后,通過轉(zhuǎn)錄組測序分析藻藍(lán)蛋白對(duì)致衰小鼠卵巢功能改善的分子機(jī)理,為臨床治療卵巢功能的研究提供基礎(chǔ)數(shù)據(jù)。實(shí)驗(yàn)以B6D2F/1品系小鼠為研究對(duì)象,分別采集8周齡小鼠卵巢作為對(duì)照組,D-半乳糖致衰小鼠卵巢作為致衰組,藻藍(lán)蛋白灌胃D-半乳糖致衰小鼠卵巢作為致衰改善組。利用轉(zhuǎn)錄組測序(RNA-Sequencing,RNA-seq)的方法對(duì)這三組不同來源的卵巢進(jìn)行轉(zhuǎn)錄組測序分析。通過基因本體(Gene Ontology,GO)分類及KEGG(Kyoto Encyclopedia of Genes and Genomes)Pathway 進(jìn)行基因差異表達(dá)分析和差異基因的功能分析,得到如下實(shí)驗(yàn)結(jié)果:(1)按照P0.05篩選差異基因,對(duì)照組和致衰組差異分析共篩選出71個(gè)差異基因;對(duì)照組和致衰改善組共篩選出56個(gè)差異基因;致衰組和致衰改善組共篩選出89個(gè)差異基因。我們集中分析了致衰組和致衰改善組差異表達(dá)基因的功能,發(fā)現(xiàn)這些差異基因主要參與調(diào)節(jié)細(xì)胞分化Sfrp2、Efnb2、Runx3、Jun)、細(xì)胞凋亡(Tnfrsf9、Btnl2、Ltf、Cdkn1α)、細(xì)胞程序性死亡(Tnfrsf9、Ltf、nt、1)、細(xì)胞因子產(chǎn)生(Tnfrsf9、Btnl2、Ltf)等與發(fā)育相關(guān)的過程。KEGG Pathway富集分析發(fā)現(xiàn),這些差異基因顯著富集到Wnt信號(hào)通路(Wnt signaling pathway)、TNF信號(hào)通路(TNF signaling pathway)、p53 信號(hào)通路(p53 signaling pathway)及 NF-κB信號(hào)通路(NF-kappa B signaling pathway)等與衰老有關(guān)的通路。(2)結(jié)合上述分析,我們篩選了 Aocl、Cd74、Cox6a2、Cyp4f18、Enpp3、Tnfrsf9、Wnt11、Zfp365等8個(gè)與衰老相關(guān)的差異基因進(jìn)行實(shí)時(shí)熒光定量分析和驗(yàn)證。與RNA-seq的結(jié)果相比,兩種方法得到的基因表達(dá)差異趨勢一致,證明了 RNA-seq結(jié)果的可靠性,并發(fā)現(xiàn)這8個(gè)基因與炎癥、癌癥、免疫相關(guān),在灌胃藻藍(lán)蛋白后,這些基因在致衰改善組中發(fā)生顯著改變,我們推測這些基因的表達(dá)變化可能對(duì)衰老卵巢的功能改善起到一定促進(jìn)作用。綜上所述,本研究通過RNA-seq分析發(fā)現(xiàn)致衰小鼠的卵巢轉(zhuǎn)錄組活動(dòng)發(fā)生較大改變,這些改變在一定程度上可以通過灌胃藻藍(lán)蛋白而恢復(fù),鑒定出的相關(guān)差異表達(dá)基因參與了免疫、炎癥及衰老有關(guān)的信號(hào)通路,提示我們藻藍(lán)蛋白可能通過這些信號(hào)通路發(fā)揮對(duì)衰老小鼠卵巢功能的改善作用,為未來卵巢功能相關(guān)研究提供了有效的靶點(diǎn)。
[Abstract]:The ovary is one of the most important organs in the female reproductive system. As women get older, their functions decline, and they experience follicle decline, oocyte quality decline, irregular menstruation, and ovarian hormone deficiency. Stopping ovulation leads to a decline in fertility, and delaying ovarian aging is an important means of increasing reproductive capacity. Phycocyanin C-Phycocyanin PCC, a form of spirulina platensis, has anti-inflammatory, antioxidant, and anti-cancer properties. A protein that improves immunity and protects the liver and kidney, because it has a variety of biological activities and has been used in many clinical studies. We have found that PC can improve ovarian function of mice induced by D-galactose, but its molecular mechanism has not been fully studied. The molecular mechanism of phycocyanin on ovarian function improvement in aging mice was analyzed by transcriptome sequencing, which provided basic data for clinical treatment of ovarian function. The ovaries of 8-week-old mice were collected as control group. The ovaries of aging mice induced by phycocyanin were treated as the improvement group of aging by intragastric administration of phycocyanin. The transcriptional sequence analysis of these three groups of ovaries from different sources was carried out by the method of transcriptional sequencing. The classification and KEGG(Kyoto Encyclopedia of the three groups of ovaries from different sources were analyzed. Genes and Genomes)Pathway was used to analyze gene differential expression and function of differentially expressed genes. The results are as follows: 1) according to P0.05, 71 differentially genes were screened in control group and senescence group, 56 genes were screened in control group and senescence improvement group, 56 genes in control group and senescence improvement group were screened out. A total of 89 differentially expressed genes were screened from aging group and senescence improvement group. We focused on the function of differentially expressed genes in senescence group and senescence improvement group. It was found that these differentially expressed genes were mainly involved in the processes of regulating cell differentiation, such as regulation of cell differentiation, Sfrp2, Efnb2, Runx3, Tnfrsf9, Btnl2, Tnfrsf9, Tnfrsf9, Tnfrsf9, Tnfrsf9, Tnfrsf9Btnl2, etc., and KEGG Pathway enrichment analysis showed that these genes were mainly involved in the processes related to development, such as the regulation of cell differentiation, Tnfrsf9, Btnl2, Tnfrsf9, Tnfrsf9, Tnfrsf9Btnl2, etc. These differentially expressed genes were significantly enriched in the Wnt signaling pathway, signaling signaling pathway, and NF- 魏 B signal pathway, NF-kappa B signaling pathway, etc., combined with the above analysis. We have screened eight different genes related to senescence, such as Aocl-CD74Cox6a2Cyp4f18, Enpp3Tnfrsf9, Wnt11Zfp365, and carried out real-time fluorescence quantitative analysis and verification. Compared with the results of RNA-seq, the difference trend of gene expression obtained by the two methods is consistent, which proves the reliability of the results of RNA-seq. It was also found that these eight genes were associated with inflammation, cancer, and immunity, and that these genes were significantly altered in the aging improvement group after oral administration of phycocyanin. We speculate that the changes of the expression of these genes may play a role in improving the function of aging ovaries. In conclusion, we found that the transcriptional activity of the ovaries in aging mice changed greatly by RNA-seq analysis. These changes can be recovered in part by oral administration of phycocyanin, and the identified differentially expressed genes are involved in signaling pathways related to immunity, inflammation and aging. These results suggest that phycocyanin may play an important role in improving the ovarian function of aging mice through these signaling pathways and provide an effective target for the future study of ovarian function.
【學(xué)位授予單位】：內(nèi)蒙古大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R711.75

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 LV Xing;LV Gao-Hong;DAI Guo-Ying;SUN Hong-Mei;XU Hui-Qin;;Food-advanced glycation end products aggravate the diabetic vascular complications via modulating the AGEs/RAGE pathway[J];Chinese Journal of Natural Medicines;2016年11期

2 Linlin Pan;Wei Gong;Yuanyuan Zhou;Xiaonuan Li;Jun Yu;Songnian Hu;;A Comprehensive Transcriptomic Analysis of Infant and Adult Mouse Ovary[J];Genomics,Proteomics & Bioinformatics;2014年05期

3 孫江宏;劉玉梅;曹統(tǒng);歐陽五慶;;激動(dòng)素對(duì)D-半乳糖致衰雌性小鼠卵巢和子宮的影響[J];生理學(xué)報(bào);2013年04期

4 ;Current understanding of ovarian aging[J];Science China(Life Sciences);2012年08期

5 祁云霞;劉永斌;榮威恒;;轉(zhuǎn)錄組研究新技術(shù):RNA-Seq及其應(yīng)用[J];遺傳;2011年11期

6 ;Next-generation sequencing technology:A technology review and future perspective[J];Science China(Life Sciences);2010年01期

7 ;Deficit of mitochondria-derived ATP during oxidative stress impairs mouse MII oocyte spindles[J];Cell Research;2006年10期

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本文編號(hào)：1650507